• 제목/요약/키워드: Nuclear localization

검색결과 309건 처리시간 0.022초

항 암태아성항원에 대한 단세포군항체의 $^{99m}Tc$ 표지법개발 및 생체분포 ($^{99m}Tc-Labeling$ of Monoclonal Antibody to Carcinoembryonic Antigen and Biodistribution)

  • 문대혁;정준기;이명철;고창순;정홍근;박재갑
    • 대한핵의학회지
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    • 제26권2호
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    • pp.380-391
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    • 1992
  • This study was designed to evaluate a direct method of $^{99m}Tc$ labeling using $\beta-mercaptoethanol$ as a reducing agent, and to investigate whether $^{99m}Tc$ labeled specific monoclonal antibody against carcinoembryonic antigen (CEA-92) can be used for the scintigraphic localization of human colon cancer xenograft. Purified CEA-92 IgG was fragmented into F $(ab')_2$ and then labeled with $^{99m}Tc$ by transchelation method using glucarate as a chelator. Labeling efficiency, immunological reactivity and in vitro stability of $^{99m}Tc$ CEA-92 F $(ab')_2$ were measured and then injected intravenously into nude mice bearing human colon cancer (SNU-C4). Scintigrams were obtained at 24 hour after injection. Then nude mice were sacrificed and the radioactivity was measured Labeling efficiency of injected $^{99m}Tc$ CEA-92 F $(ab')_2$, immunoreative fraction and in vitro stability at 24 hour of injected $^{99m}Tc$ CEA-92 F $(ab')_2$ was 45.2%, 32.8% and 57.4%, respectively. At 24 hour after injection, % ID/g in kidney (46.77) showed high uptake, but %ID/g in tumor (1.65) was significantly higher than spleen (0.69), muscle (0.16), intestine (0.45), stomach (0.75), heart (0.48) and blood (0.45). There was no significant difference between tumor and liver (1.81). Tumor contrast as quantitated by tumor to blood ratio of $^{99m}Tc$ CEA-92 F $(ab')_2$ was increased significantly (p<0.005) until 24 hours (3.70), and there was no statistical differece from tumor to blood ratio of I-131 CEA-92 F $(ab')_2$. The scintigram demonstrated localization of radioactivity over transplanted tumor, but significant background radioactivity was also noted over kidney and abdomen. It is concluded that CEA-92 F $(ab')_2$ can be labeled with $^{99m}Tc$ by a direct transchelation method using $\beta-mercaptoethanol$ as a reducing agent and $^{99m}Tc$ labeled CEA-92 F $(ab')_2$ can be used for the scintigraphic localization of human colon cancer xenograft in nude mice model.

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IMACIS-1을 이용한 위장관 종양의 방사면역신티그램 (Radioimmunoscintigraphy Using IMACIS-1 in Gastrointestinal Cancer)

  • 손형선;김춘열;박용휘
    • 대한핵의학회지
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    • 제24권1호
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    • pp.29-36
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    • 1990
  • Most of the diagnostic methods currently used for the detection of neoplastic masses provide indirect evidence. To obtain greater specificity in the interpretation of neoplasias by in vivo methods, the immunological approach appears to be most promising. Two problems that interfered with progress in this field were the lack of tumor specific antigen and the lack of well-defined and reproducible antibodies. To improve the sensitivity and specificity of radioimmunoscintigraphy as a technique for tumor localization, the use of monoclonal antibodies, fragments of antibodies and single photon emission computerized tomography (SPECT) are reasonable. The obvious advantages of monoclonal antibodies are their homogeneity, their specificity for the immunizing antigen and the reaction with a single determinant-thus no large immunecomplexes with antigen are formed. Monoclonal antibody technique has recently provided an opportunity to reevaluate the role of nuclear medicine for the diagnosis of malignant diseases by using the immunological approach. Out first results by means of radioimmunoscintigraphy of CEA and CA 19-9 producing tumors using a cocktail of fragments F $(ab')_2$, of mocolonal antibodies to CA 19-9 and CEA labeled with $^{131}I$ (IMACIS-1) are reported. The aims of this investigation was to evaluate the role of immunoscintigraphy in patients with colorectal and other cancers for diagnosis of local recurrences and metastasis. This report contains results of the first 8 colorectal and pancreas cancer patients with the elevation of the level of serum CEA and/or CA 19-9. IMACIS-1 was injected intravenously during 30 minutes in 100 ml saline solution after skin test. Planar scintigrams were recorded 3, 5 and 7 days after the injection of the IMACIS-1. Anterior, lateral and posterior views of the liver as well as anterior and posterior views of the pelvis were obtained in each patients as an $^{131}I-antibody$ image. We were able to localize exactly the malignant process with the double-nuclide double-compound $^{99m}Tc\;^{131}I$ (Tc+l) scintigrams. In Tc & I double-nuclide scintigraphy, computer subtraction display provided more clear localization of the tumor. We compared the results of radioimmunoscintigraphy with CT, ultrasonograms, conventional scintigrams. The results were as follows: 1) The sensitivity and specificity of radioimmunoscintigraphy using the fragments $F(ab')_2$ of the cocktails of CEA and CA 19-9 monoclonal antibodies were 80% and 100% respectively. 2) Tumor detection rate was not proportionated to the level of serum tumor markets. 3) Second tracer technique was essential for tumor localization as an anatomic landmark using double-nuclide scintigraphy. 4) A slow infusion of the antibodies was necessary to prevent the formation of large immune complexes. 5) Tumor/non-tumor radioactivity was most elevated at 7 days delayed imaging. 6) Using planar scintigraphic technique of $^{131}I$ labeled monoclonal antibodies are possible for imaging most of the tumors.

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Expression and Localization of Heat Shock Protein 70 in Frozen-Thawed IVF and Nuclear Transfrred Bovine Embryos

  • Park, Y.J;S.J Song;J.T Do;B.S Yoon;Kim, A.J;K.S Chung;Lee, H.T
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2002년도 국제심포지엄
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    • pp.78-78
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    • 2002
  • The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

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Protein transduction of an antioxidant enzyme: subcellular localization of superoxide dismutase fusion protein in cells

  • Kim, Dae-Won;Kim, So-Young;Lee, Hwa;Lee, Yeum-Pyo;Lee, Min-Jung;Jeong, Min-Seop;Jang, Sang-Ho;Park, Jin-Seu;Lee, Kil-Soo;Kang, Tae-Cheon;Won, Moo-Ho;Cho, Sung-Woo;Kwon, Oh-Shin;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • 제41권2호
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    • pp.170-175
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    • 2008
  • In protein therapy, it is important for exogenous protein to be delivered into the target subcellular localization. To transduce a therapeutic protein into its specific subcellular localization, we synthesized nuclear localization signal (NLS) and membrane translocation sequence signal (MTS) peptides and produced a genetic in-frame SOD fusion protein. The purified SOD fusion proteins were efficiently transduced into mammalian cells with enzymatic activities. Immunofluorescence and Western blot analysis revealed that the SOD fusion proteins successfully transduced into the nucleus and the cytosol in the cells. The viability of cells treated with paraquat was markedly increased by the transduced fusion proteins. Thus, our results suggest that these peptides should be useful for targeting the specific localization of therapeutic proteins in various human diseases.

Construction of a Novel Recombinant Bombyx mori Nuclear Polyhedrosis Virus Producing the Fluorescent Polyhedra

  • Kang, Seok-Woo;Yun, Eun-Young;Woo, Soo-Dong;Goo, Tae-Won;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • 제3권1호
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    • pp.75-81
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    • 2001
  • We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

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가우시안 포락선 선형 첩 신호의 순시 주파수 추정을 통한 원전 내 계측 케이블의 고장점 진단 연구 (Instantaneous Frequency Estimation of the Gaussian Enveloped Linear Chirp Signal for Localizing the Faults of the Instrumental Cable in Nuclear Power Plant)

  • 이춘구;박진배;윤태성
    • 전기학회논문지
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    • 제62권7호
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    • pp.987-993
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    • 2013
  • Integrity of the control and instrumental cables in nuclear power plant is important to maintain the stability of the nuclear power plants. In order to diagnose the integrity of the cables, the diagnostic methods based on reflectometry have been studied. The reflectometry is a non-destructive method and it is applicable to diagnose the live cables. We introduce a Gaussian enveloped linear chirp reflectometry to diagnose the cables in the nuclear power plants. In this paper, we estimate the instantaneous frequency of the Gaussian enveloped linear chirp signal by using the weighted robust least squares filtering to localize the impedance discontinuities in the class 1E instrumental cable.

CRM1 inhibitor S109 suppresses cell proliferation and induces cell cycle arrest in renal cancer cells

  • Liu, Xuejiao;Chong, Yulong;Liu, Huize;Han, Yan;Niu, Mingshan
    • The Korean Journal of Physiology and Pharmacology
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    • 제20권2호
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    • pp.161-168
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    • 2016
  • Abnormal localization of tumor suppressor proteins is a common feature of renal cancer. Nuclear export of these tumor suppressor proteins is mediated by chromosome region maintenance-1 (CRM1). Here, we investigated the antitumor effects of a novel reversible inhibitor of CRM1 on renal cancer cells. We found that S109 inhibits the CRM1-mediated nuclear export of RanBP1 and reduces protein levels of CRM1. Furthermore, the inhibitory effects of S109 on CRM1 is reversible. Our data demonstrated that S109 significantly inhibits proliferation and colony formation of renal cancer cells. Cell cycle assay showed that S109 induced G1-phase arrest, followed by the reduction of Cyclin D1 and increased expression of p53 and p21. We also found that S109 induces nuclear accumulation of tumor suppressor proteins, Foxo1 and p27. Most importantly, mutation of CRM1 at Cys528 position abolished the effects of S109. Taken together, our results indicate that CRM1 is a therapeutic target in renal cancer and the novel reversible CRM1 inhibitor S109 can act as a promising candidate for renal cancer therapy.

Low-noise reconstruction method for coded-aperture gamma camera based on multi-layer perceptron

  • Zhang, Rui;Tang, Xiaobin;Gong, Pin;Wang, Peng;Zhou, Cheng;Zhu, Xiaoxiang;Liang, Dajian;Wang, Zeyu
    • Nuclear Engineering and Technology
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    • 제52권10호
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    • pp.2250-2261
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    • 2020
  • Accurate localization of radioactive materials is crucial in homeland security and radiological emergencies. Coded-aperture gamma camera is an interesting solution for such applications and can be developed into portable real-time imaging devices. However, traditional reconstruction methods cannot effectively deal with signal-independent noise, thereby hindering low-noise real-time imaging. In this study, a novel reconstruction method with excellent noise-suppression capability based on a multi-layer perceptron (MLP) is proposed. A coded-aperture gamma camera based on pixel detector and coded-aperture mask was constructed, and the process of radioactive source imaging was simulated. Results showed that the MLP method performs better in noise suppression than the traditional correlation analysis method. When the Co-57 source with an activity of 1 MBq was at 289 different positions within the field of view which correspond to 289 different pixels in the reconstructed image, the average contrast-to-noise ratio (CNR) obtained by the MLP method was 21.82, whereas that obtained by the correlation analysis method was 5.85. The variance in CNR of the MLP method is larger than that of correlation analysis, which means the MLP method has some instability in certain conditions.