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PAMAM Dendrimer Conjugated with N-terminal Oligopeptides of Mouse Fibroblast Growth Factor 3 as a Novel Gene Carrier

  • Jung, Jinwoo;Lee, Jeil;Kim, Tae-Hun;Yang, Bong Suk;Lee, Eunji;Kim, Youn-Joong;Park, Jong-Sang;Choi, Joon Sig
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1036-1042
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    • 2014
  • In this study, we introduced the RRLR oligopeptide sequences on the surface of polyamidoamine (PAMAM) dendrimer and characterized the physical properties and gene carrier activity of the novel polymer using HEK 293, NIH3T3, and HeLa cells. The RRLR peptide sequences were derived from a mouse fibroblast growth factor 3 (FGF3) protein containing a bipartite NLS motif. The entire sequence of FGF3 is RLRRDAGGRGGVYEHLGGAPRRRK and it has two functional sequences RLRR and RRRK at N-terminus and C-terminus, respectively. In particular, PAMAM G4-RRLR conferred enhanced transfection efficiency and lower cytotoxicity compared with those of PEI 25 kDa, PAMAM G4-R, and PAMAM G4 in various cell lines. These results suggest that the introduction of N-terminal oligopeptides of FGF3 on the surface of PAMAM holds promise as an effective non-viral gene delivery carrier for gene therapy.

Protective Effect of Aster tataricus L. Extract on the Dermal Cytotoxicity Induced by Sodium Bromate, Oxidant of Hair Dye

  • Chung, Jung-Hwa;Lee, Gyoung-Wan;Seo, Young-Mi
    • Biomedical Science Letters
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    • v.25 no.4
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    • pp.348-356
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    • 2019
  • This study evaluated the dermal cytotoxicity of sodium bromate (NaBrO3) and the protective effect of Aster tataricus L. (AT) extract against NaBrO3-induced cytotoxicity in the cultured NIH3T3 fibroblasts. For this study, it was done the antioxidative effects such as electron donating (ED) activity and lipid peroxidation (LP) activity as well as cell viability. NaBrO3 significantly decreased cell viability in a dose-dependent manner and its XTT50 value was measured at a concentration of 54.4 μM in these cultures. The cytotoxicity of NaBrO3 was determined as highly-toxic by Borenfreund and Puerner's toxic criteria. The quercetin, antioxidant significantly increased cell viability against NaBrO3-induced cytotoxicity. Regarding the protective effect of Aster tataricus (AT) L. extract on NaBrO3-induced cytotoxicity, AT extract significantly increased the cell viability, the ED ability and the inhibitory ability of LP. From these findings, it suggested that the oxidative stress is involved in the cytotoxicity of NaBrO3, and AT extract effectively protected NaBrO3-induced cytotoxicity by antioxidative effects. Conclusively, the natural component like AT extract may be a putative therapeutic agent for the diminution or treatment of the cytotoxicity correlated with oxidative stress like hair dye component, NaBrO3.

Green Fluorescent Protein-reporter Mammalian One-hybrid System for Identifying Novel Transcriptional Modulators for Human $p14^{ARF}$ Tumor Suppressor Gene

  • Lee, Hye Jin;Yang, Dong Hwa;Yim, Tae Hee;Rhee, Byung Kirl;Kim, Jung-Wook;Lee, Jungwoon;Gim, Jin Bae;Kim, JungHo
    • Animal cells and systems
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    • v.6 no.4
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    • pp.317-322
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    • 2002
  • To improve conventional yeast one-hybrid screening, we have developed an efficient mammalian one-hybrid system that allows rapid isolation of com-plementary DNAs which are able to induce human p14$^{ARF}$. tumor suppressor gene. A 1.5 kb promoter region of p14$^{ARF}$ was fused to EGFP to generate ARF promoter-EGFP reporter vector. This reporter plasmid was stably trans-fected into NIH3T3 cells for generation of reporter cell line. When the reporter cell line was infected with E2F-1 together with excess amounts of empty vector, the cells that received the positive modulator were readily identifiable by green fluorescence using FACS. The GFP-positive cells were cloned directly from the cultured cells and expanded in bulk culture. The genomic DNAs from GFP-positive cells were prepared and the CDNA insert in integrated retroviral genome was recovered by PCR using primers annealing to the retroviral vector sequences flanking the insert-cloning site. This system should be useful for efficient screening of expression CDNA libraries in mammalian cells to identify novel upstream regulators for spe-cific genes by one-hybrid interaction.ion.

The Suppressive Effect of Pueraria lobata Root Extract and Its Biotransformed Preparation against Skin Wrinkle Formation

  • Koo, Hyun Jung;Lee, SungRyul;Kang, Se Chan;Kwon, Jung Eun;Lee, Da Eun;Choung, Eui-Su;Lee, Jong-Sub;Lee, Jin Woo;Park, Yuna;Sim, Dong Soo;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
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    • v.30 no.3
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    • pp.272-279
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    • 2017
  • EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to $1000{\mu}g/ml$ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of $50{\mu}g/ml$ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of $50{\mu}g/ml$. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

The effect of bacterial cellulose membrane compared with collagen membrane on guided bone regeneration

  • Lee, So-Hyoun;Lim, Youn-Mook;Jeong, Sung In;An, Sung-Jun;Kang, Seong-Soo;Jeong, Chang-Mo;Huh, Jung-Bo
    • The Journal of Advanced Prosthodontics
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    • v.7 no.6
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    • pp.484-495
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    • 2015
  • PURPOSE. This study was to evaluate the effects of bacterial cellulose (BC) membranes as a barrier membrane on guided bone regeneration (GBR) in comparison with those of the resorbable collagen membranes. MATERIALS AND METHODS. BC membranes were fabricated using biomimetic technology. Surface properties were analyzed, Mechanical properties were measured, in vitro cell proliferation test were performed with NIH3T3 cells and in vivo study were performed with rat calvarial defect and histomorphometric analysis was done. The Mann-Whitney U test and the Wilcoxon signed rank test was used (${\alpha}<.05$). RESULTS. BC membrane showed significantly higher mechanical properties such as wet tensile strength than collagen membrane and represented a three-dimensional multilayered structure cross-linked by nano-fibers with 60 % porosity. In vitro study, cell adhesion and proliferation were observed on BC membrane. However, morphology of the cells was found to be less differentiated, and the cell proliferation rate was lower than those of the cells on collagen membrane. In vivo study, the grafted BC membrane did not induce inflammatory response, and maintained adequate space for bone regeneration. An amount of new bone formation in defect region loaded with BC membrane was significantly similar to that of collagen membrane application. CONCLUSION. BC membrane has potential to be used as a barrier membrane, and efficacy of the membrane on GBR is comparable to that of collagen membrane.

A Clinical Study of Decrease Appetite Effects by Aromatherapy Using Foeniculum vulgare Mill(Fennel) to Female Obese Patients (여성비만환자에게 소회향(小茴香) 정유를 사용한 향기요법의 식욕억제효과에 관한 임상적 연구)

  • Kim, Se-Jong;Kim, Kil-Soo;Choi, Young-Min;Kang, Byung-Gab;Yoon, Yoo-Sik;Oh, Min-Suk;Yoon, Il-Ji;Shin, Seung-Uoo
    • Journal of Korean Medicine for Obesity Research
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    • v.5 no.1
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    • pp.9-20
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    • 2005
  • Objectives : This study was performed to investigate the effect of Aromatherapy using Foeniculum vulgare Mill(Fennel) for decrease in the appetite of female obese patients. Methods : We analyzed 32 obese patients who had been hospitalized in Kirin Oriental Hospital from October 1, 2004 to January 30, 2005. The patient were classified in two groups, aroma inhalation group(IH group), and non-inhalation group(NIH group). Test was applied 2 times a week(Inhalation and Non-inhalation test) for 4 weeks. The effect of both groups was analyzed by using VAS scores about appetite. The scores were checked before inhalation, and 0, 15, 30, 45, 60 minutes after inhalation. Results : The VAS appetite scores decreased more significantly in inhalation group than non-inhalation group on every checked time(15, 30, 45, 60 minutes after inhalation)(p<0.05). And VAS scores of aroma inhalation group decreased more significantly on every checked time(0, 15, 30, 45, 60 minutes after inhalation) than pre-inhalation(p<0.05), and morbid obesity group is more sensitive than obesity group shortly after the inhalation(0 minute after inhalation)(p<0.05). Conclusions : In this study, we can conclude that aromatherapy using Foeniculum vulgare Mill(Fennel) has an effect on decreasing appetite of female obese patients.

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Cytotoxic Effects of Partially Purified Substances from Bacillus polyfermenticus SCD Supernatant toward a Variety of Tumor Cell tines

  • Chang, Kyung-Hoon;Park, Jun-Seok;Choi, Jae-Hoon;Kim, Cheon-Jei;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.163-166
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    • 2007
  • The cytotoxic effects of partially purified substances from Bacillus polylfermenticus SCD toward a variety tumor cell lines were studied. Cytotoxic activity was determined with regard to the A549 (human lung carcinoma), AGS (human stomach adenocarcinoma), DLD-1 (human colon adenocarcinoma), HEC-1-B (human uterus adenocarcinoma), SW-156 (human kidney carcinoma), and NIH/3T3 (murine normal fibroblast) cell lines using the MTT assay. Cytotoxic substances were partially purified through Diaion HP-20 columns and extracted with methanol or other organic solvents (n-hexane, chloroform, ethylacetate, and butanol). B. polyfermenticus SCD supernatant showed up to 60% inhibition of cell viability fer all five human cancer cell lines tested. When treated with 10 mg/mL of n-hexane, chloroform, ethylacetate, and butanol extract, HEC-1-B cells showed a 25,62,35, and 63% rate of inhibition respectively, and AGS cells showed a 72, 61, 44, and 67% rate of inhibition, respectively. At a concentration of 10 mg/mL, 100% methanol Diaion HP-20 extracts showed inhibition rates of 97.0% toward A-549 cells, 98.1% toward AGS cells, 81.6% toward DLD-1 cells, 83.5% toward HEC-1-B cells, and 92.7% toward SW-156 cells. These results indicate that partially purified fractions from B. polyfermenticus SCD have the potential to inhibit not only colon cancer cells, but also lung, stomach uterus, and kidney cancer cells. Further studies are needed to characterize the cytotoxic substances released in B. polyfermenticus SCD cultures.

Antitumor Effects of Kluyveromyces marxianus TFM-7 Isolated from Kefir

  • Lee, Hyun-Jung;Nam, Bo-Ra;Kim, Jin-Man;Kim, Ji-Yeon;Paik, Hyun-Dong;Kim, Chang-Han
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.133-137
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    • 2007
  • The Strain TFM-7, Which has an antitumor effect, was isolated from Kefir and identified based on analysis using the API 50 CHL kit and 265 rDNA sequencing. Strain TFM-7 was confirmed to belong to the genus Kluyveromyces. Analysis of the 265 rDNA nucleotide sequences found strain TFM-7 to be related to Kluyveromyces marxianus. NRRL Y-828IT. K. marxianus. TFM-7 was cultured with potato dektrose broth medium at $27^{\circ}C$ for 72 hr, and its inhibition effects on the proliferation of seven tumor cell lines and a normal cell line were assessed using the MTT assay. The antitumor effects and growth characteristics of K. marxianus TFM-7 were investigated during a culture period of 7 days. By the $3^{rd}\;day$, K. marxianus TFM-7 showed a dry cell weight 2.39 g/L, a pH of 4.39, an ethanol content of 0.89%, and an inhibition effect on the proliferation of seven tumor cell lines above 50%, except for A-549 tumor cell line. K. marxianus TFM-7 was the most effective at inhibiting the growth of Hep-2 cell line among all tumor cell lines tested. Growth inhibition of a normal cell line, NIH/3T3, was less than 35%, suggesting a decreased level of cytotoxicity toward normal cells. These results indicate that K. marxianus TFM-7 may have used as a yeast strain with antitumor activity.

Identification of the Antidepressant Vilazodone as an Inhibitor of Inositol Polyphosphate Multikinase by Structure-Based Drug Repositioning

  • Lee, Boah;Park, Seung Ju;Lee, Seulgi;Park, Seung Eun;Lee, Eunhye;Song, Ji-Joon;Byun, Youngjoo;Kim, Seyun
    • Molecules and Cells
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    • v.43 no.3
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    • pp.222-227
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    • 2020
  • Inositol polyphosphate multikinase (IPMK) is required for the biosynthesis of inositol phosphates (IPs) through the phosphorylation of multiple IP metabolites such as IP3 and IP4. The biological significance of IPMK's catalytic actions to regulate cellular signaling events such as growth and metabolism has been studied extensively. However, pharmacological reagents that inhibit IPMK have not yet been identified. We employed a structure-based virtual screening of publicly available U.S. Food and Drug Administration-approved drugs and chemicals that identified the antidepressant, vilazodone, as an IPMK inhibitor. Docking simulations and pharmacophore analyses showed that vilazodone has a higher affinity for the ATP-binding catalytic region of IPMK than ATP and we validated that vilazodone inhibits IPMK's IP kinase activities in vitro. The incubation of vilazodone with NIH3T3-L1 fibroblasts reduced cellular levels of IP5 and other highly phosphorylated IPs without influencing IP4 levels. We further found decreased Akt phosphorylation in vilazodone-treated HCT116 cancer cells. These data clearly indicate selective cellular actions of vilazodone against IPMK-dependent catalytic steps in IP metabolism and Akt activation. Collectively, our data demonstrate vilazodone as a method to inhibit cellular IPMK, providing a valuable pharmacological agent to study and target the biological and pathological processes governed by IPMK.

ELF 3D Magnetic Field and Eddy Current Calculation of Human Body Around Transmission Lines (송전선로 주변의 3차원 자기장 및 인체 유도 와전류 계산)

  • Myeong, Seong-Ho;Lee, Dong-Il;Sin, Gu-Yong;Han, In-Su;Park, Jong-Geun
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.10
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    • pp.485-491
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    • 2002
  • Since Wertheimer and Leeper reported possible adverse health effects of magnetic field in 1979, worldwide researches on this issue have been conducted. More recently, the U.S. Congress instructed the NIEHS (National Institute of Environmental Health Sciences), NIH (National Institute of Health) and DOE (Department of Energy) to direct and manage EMF RAPID (Electric and Magnetic Fields Research and Public Information Dissemination) program aimed at providing scientific evidence to clarify the potential for health risks from exposure to extremely low frequency electric and magnetic fields(ELF-EMF). Although they concluded that the scientific evidence suggesting adverse health risks of ELF-EMF is weak, the exposure to ELF-EMF cannot be recognized as entirely safe. Therefore, the purpose of this article is to describe magnetic field 3-D calculation and to evluate eddy current of human body compare to international guide line recognized one of the basic problems. In open boundary problem, Magnetic field using FEM is not advantageous in the point of the division of area and the proposition of the fictitious boundary. Therefore, we induced the analytic equation of magnetic field calculations so but the finite line segment based on Biot-Savarts law Also, Eddy currents induced due to ELF-EMF magnetic field are computed. To calculate induced currents, impedance method is used in this paper, An example model of human head with resolution of 1.27cm is used. In this paper, We evaluate the magnetic field and eddy current of human head around 765 kV transmission lines compare to international guide line.