• Title/Summary/Keyword: Imaging probes

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Screening and Characterization of a Novel RNA Aptamer That Specifically Binds to Human Prostatic Acid Phosphatase and Human Prostate Cancer Cells

  • Kong, Hoon Young;Byun, Jonghoe
    • Molecules and Cells
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    • v.38 no.2
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    • pp.171-179
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    • 2015
  • Prostatic acid phosphatase (PAP) expression increases proportionally with prostate cancer progression, making it useful in prognosticating intermediate to high-risk prostate cancers. A novel ligand that can specifically bind to PAP would be very helpful for guiding prostate cancer therapy. RNA aptamers bind to target molecules with high specificity and have key advantages such as low immunogenicity and easy synthesis. Here, human PAP-specific aptamers were screened from a 2'-fluoropyrimidine (FY)-modified RNA library by SELEX. The candidate aptamer families were identified within six rounds followed by analysis of their sequences and PAP-specific binding. A gel shift assay was used to identify PAP binding aptamers and the 6N aptamer specifically bound to PAP with a Kd value of 118 nM. RT-PCR and fluorescence labeling analyses revealed that the 6N aptamer bound to PAP-positive mammalian cells, such as PC-3 and LNCaP. IMR-90 negative control cells did not bind the 6N aptamer. Systematic minimization analyses revealed that 50 nucleotide sequences and their two hairpin structures in the 6N 2'-FY RNA aptamer were equally important for PAP binding. Renewed interest in PAP combined with the versatility of RNA aptamers, including conjugation of anti-cancer drugs and nano-imaging probes, could open up a new route for early theragnosis of prostate cancer.

Use of DNA-Specific Anthraquinone Dyes to Directly Reveal Cytoplasmic and Nuclear Boundaries in Live and Fixed Cells

  • Edward, Roy
    • Molecules and Cells
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    • v.27 no.4
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    • pp.391-396
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    • 2009
  • Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

Acoustic Power Measurement System of Array Probes for Ultrasonic Diagnostic Equipment Using Radiation Force Balance Methods (방사힘 측정법을 이용한 초음파 진단장치용 배열 탐침자의 음향파워 측정시스템)

  • Yun, Yong-Hyeon;Jho, Moon-Jae;Kim, Yong-Tae;Lee, Myoung-Ho
    • The Journal of the Acoustical Society of Korea
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    • v.29 no.6
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    • pp.355-364
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    • 2010
  • Considering biological safety, it is very important to measure acoustic power from ultrasonic array probe for diagnostic ultrasound imaging applications. In this paper, to measure acoustic power from each element on array probe for ultrasonic diagnostic equipment, we reconstruct and automate the acoustic power measurement system. The acoustic power from linear, phased and curved array were measured and analyzed. As a result of measurement, the effects caused by directivity of sound beam from curved array were founded. To remove these effects, we developed and applied the correction model. The proposed system is useful to evaluate characteristics of the acoustical output power of array probe.

In vivo comparison of Lu-177-labeled phosphonate compounds as potential agents for bone pain palliation in rodents

  • Chang, Young Soo;Lee, Yun-Sang;Kim, Young Ju;Jeong, Jae Min
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.1 no.1
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    • pp.53-61
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    • 2015
  • Lutetium-177 ($T_{1/2}=6.71day$) is an adequate radionuclide for therapy, which has both beta emission ($E_{max}=497keV$) for therapeutic effect and gamma emission (113 and 208 keV) for imaging. $^{177}Lu$ labeled ethylenediamine-N,N,N',N'-tetrakis (methylene phosphonic acid) (EDTMP) and 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraaminomethylenephosphonate (DOTMP) have been proposed as radiopharmaceuticals for bone pain palliation. In this study, we compared radiochemistry and biodistribution of $^{177}Lu$-EDTMP and $^{177}Lu$-DOTMP. EDTMP and DOTMP were synthesized, and 1 mg of each was labeled with $^{177}Lu$ at pH 7~8 with high efficiency (>98%). For comparative biodistribution studies, $^{177}Lu$-EDTMP or $^{177}Lu$-DOTMP were injected into ICR-mice through tail vein, and then biodistribution data were obtained as percentages of injected dose per gram of tissue (% ID/g). Urine excretions of both agents in mice were checked for 7 days. Rat images were also obtained after injection of $^{177}Lu$-EDTMP or $^{177}Lu$-DOTMP. $^{177}Lu$-DOTMP (100% at 1 min) showed faster labeling than $^{177}Lu$-EDTMP (100% at 30 min). Both of them were stable at least for 21 days at room temperature. High bone uptakes were found for both $^{177}Lu$-EDTMP and $^{177}Lu$-DOTMP: 38.0 and 34.1% ID/g at 3 hr, respectively; and 33.2 and 18.8% ID/g at 7 day, respectively. Rapid excretions to urine were found for both agents ($^{177}Lu$-EDTMP: 56%, $^{177}Lu$-DOTMP: 63% at 1 day). Other organs showed very low uptakes. Rat images of both $^{177}Lu$-EDTMP and $^{177}Lu$-DOTMP showed high bone uptakes and low soft tissue uptakes. In conclusion, both $^{177}Lu$-EDTMP and $^{177}Lu$-DOTMP showed high potential as bone pain palliation agents. $^{177}Lu$-EDTMP showed higher bone uptake and slower bone clearance in mice than those of $^{177}Lu$-DOTMP.

Development of an exclusive column method for 82Sr/82Rb generator using a 100 MeV proton linear accelerator of KOMAC

  • Kye-Ryung Kim;Yeong Su Ha;Sang-Pil Yoon;Yeon-ji Lee;Yong-Sub Cho;Hyeongi Kim;Sang-Jin Han;Jung Young Kim;Kyo Chul Lee;Jin Su Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.7 no.2
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    • pp.119-125
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    • 2021
  • 82Sr for 82Rb generator was produced through the irradiation of the proton beam on the nat.RbCI target at the target irradiation facility installed at the end of the Rl-dedicated beamline of the 100 MeV proton linear accelerator of KOMAC (Korea Multi-purpose Accelerator Complex). The average current of the proton beam was 1.2 µA for irradiation time of 150 min. For the separation and purification of the 82Sr from nat.RbCI irradiated, Chelex-100 resin was used. The activities of 82Sr in the irradiated nat.RbCI target solution and after purification were 45.29 µCi and 43.4 µCi, respectively. The separation and purification yield was 95.8%. As an adsorbent to be filled in the generator for 82Sr adsorption hydrous tin oxide was selected. The adsorption yield of 82Sr into the generator adsorbent was > 99 %, and the total amount of 82Sr adsorbed to the generator was 21.6 µCi as of the day of the 82Rb elution experiment. When the elution amount was 22 mL, the maximum82Rb elution yield was 93.3%, and the elution yield increased as the flow rate increased. After the eluted 82Rb was filled in the correction phantom of the small PET for animals, a PET image was taken. The image scan time was set to 5 min, and the phantom PET image was successfully obtained. As results of impurity analysis on eluted 82Rb using ICP-MS, nat.Rb stable isotopes that compete in vivo of 82Rb were identified as undetected levels and were determined to be No-Carrier-Added (NCA).

Thin and superthin perforator flap elevation based on preoperative planning with ultrahigh-frequency ultrasound

  • Visconti, Giuseppe;Bianchi, Alessandro;Hayashi, Akitatsu;Cina, Alessandro;Maccauro, Giulio;Almadori, Giovanni;Salgarello, Marzia
    • Archives of Plastic Surgery
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    • v.47 no.4
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    • pp.365-370
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    • 2020
  • The ability to directly harvest thin and superthin perforator flaps without jeopardizing their vascularity depends on knowledge of the microsurgical vascular anatomy of each perforator within the subcutaneous tissue up to the dermis. In this paper, we report our experience with ultrahigh-frequency ultrasound (UHF-US) in the preoperative planning of thin and superthin flaps. Between May 2017 and September 2018, perforators of seven patients were preoperatively evaluated by both ultrasound (using an 18-MHz linear probe) and UHF-US (using 48- and 70-MHz linear probes). Thin flaps (two cases) and superthin flaps (five cases) were elevated for the reconstruction of head and neck oncologic defects and lower limb traumatic defects. The mean flap size was 6.5×15 cm (range, 5×8 to 7.5×23 cm). No complications occurred, and all flaps survived completely. In all cases, we found 100% agreement between the preoperative UHF-US results and the intraoperative findings. The final reconstructive outcomes were considered satisfactory by both the surgeon and the patients. In conclusion, UHF-US was found to be very useful in the preoperative planning of thin and superthin free flaps, as it allows precise anticipation of very superficial microvascular anatomy. UHF-US may represent the next frontier in thin, superthin, and pure skin perforator flap design.

Recent Progress in Multiplexed Detection of Biomarkers Based on Quantum Dots (양자점 기반 다중 바이오마커 검출법의 연구동향)

  • Kim, Yerin;Choi, Yu Rim;Kim, Bong-Geun;Na, Hyon Bin
    • Applied Chemistry for Engineering
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    • v.33 no.5
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    • pp.451-458
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    • 2022
  • Semiconductor quantum dots (QDs) are optical probes with excellent fluorescence properties. Therefore, they have been applied to various bio-medical imaging techniques and biosensors. Due to the unique optical characteristics of wide absorption and narrow fluorescence energy bands, multiple types of signals can be generated by the combination of fluorescence wavelengths from different QDs, which enables the simultaneous detection of more than two biomarkers. In this review, the advantages and applications of QDs and QD nanobeads (QBs) in multiple biomarker assays were described, and new developments or improvements in multiplexed biomarker detection techniques were summarized. In particular, recent reports were summarized, focusing on the design strategies in immunoassay construction and signal transducing materials for fluorescence-linked immunosorbent assays using QDs and immunochromatographic assays using QBs. New detection platforms will be developed for early diagnosis of diseases and other fields if multiplexed detection technologies of excellent accuracy and sensitivity are combined with artificial intelligence algorithms.

In Vitro Uptakes of Radiolabeled IVDU and IVFRU in Herpes Simplex Virus Type-1 Thymidine Kinase (HSV1-tk) Gene Transduced Morris Hepatoma Cell Line (단순 헤르페스 제 1형 티미딘 키나제 유전자 이입 간암세포주에서 방사표지 IVDU와 IVFRU의 섭취 평가)

  • Lee, Tae-Sup;Choi, Tae-Hyun;Ahn, Soon-Hyuk;Woo, Kwang-Sun;Jeong, Wee-Sup;Kwon, Hee-Chung;Awh, Ok-Doo;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.62-73
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    • 2004
  • Purpose: The herpes simplex virus type 1 thymidine kinase gene(HSV1-tk) is an attractive candidate as a reporter gene in noninvasive reporter gene monitoring system. The HSV1-tk gene was chosen as a reporter gene, because it has been extensively studied, and there are appropriate reporter probes, substrates of HSV1-tk gene product, to apply for HSV1-tk gene imaging. We used radiolabeled 5-iodovinyl-2'-deoxyuridine (IVDU) and 5-iodovinyl-2'-fluoro-2'-deoxyuridine (IVFRU) as reporter probes for HSV1-tk gene monitoring system. Materials and Methods: We prepared HSV1-tk gene transduced Morris hepatoma cell line using retroviral vector, MOLTEN containing HSV1-tk gene. And we confirmed the HSV1-tk gene expression by Northern blotting and Western blotting. We compared in vitro uptakes of radioiodinated IVDU and IVFRU to monitor HSV1-tk gene expression in Morris hepatoma cell line (MCA) and HSV1-tk gene tranduced MCA (MCA-tk) cells until 480 minutes. We also peformed correlation analysis between percentage of HSV1-tk gene tranduced MCA cell % (MCA-tk%) and uptakes of radiolabeled IVDU or IVFRU. Results: MCA-tk cell expressed HSV1-tk mRNA and HSV1-TK protein. Two compounds showed minimal uptake in MCA, but increased uptake was observed in MCA-tk. IVDU showed 4-fold higher accumulation than IVFRU at 480 min in MCA-tk (p<0.01). Both IVDU and IVFRU uptake were linearly correlated ($R^2>0.96$) with increasing MCA-tk%. Conclusion: The radiolabeld IVDU and IVFRU showed higher specific accumulation in retrovirally HSV1-tk gene transfected Morris hepatoma cell line. Both IVDU and IVFRU could be used as good substrates for evaluation of HSV1-tk gene expression.

Recent Perspectives on Oncoplastic Breast Surgery in Korea (우리나라의 종양성형학적 유방암 수술에 대한 최신 동향)

  • Kang, Taewoo
    • Journal of Life Science
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    • v.30 no.6
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    • pp.563-569
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    • 2020
  • Oncoplastic surgery (OPS) combines oncologically safe tumor resection with aesthetically satisfying reconstruction of defects using established plastic surgery techniques. OPS is characterized by initial excision as extensive as is beneficial for oncological safety, and, once sufficient resection is complete, displacement or replacement techniques are selected based on remnant volume. The size of the lesion and the individual patient are important factors when considering the appropriate approach, and when pre-operative imaging, including MRI, is used to determine the technique, the complete removal of cancer cells by permanent pathology is essential. A frozen section is used during the operation to reduce the reoperation rate, but it is difficult to cover the entire margin surface theoretically and even harder in practice. A recent report about adequate margins has empowered OPS in its oncological safety. Considering the patients to whom each modality could be applied, basic breast volume is an important factor, and this is influenced by ethnic differences. In Europe or the US, for example, the average breast size is 36D (600 ㎤) and reduction mammoplasty is predominantly used. However, the average size of patients in our institution is 33A (300 ㎤), and so quite different approaches are selected in most cases. New techniques involving radiofrequency and fluorescence have been proposed as safe and easily accessible ways of reducing complications.

Current Status of the KMTNet Active Nuclei Variability Survey (KANVaS)

  • Kim, Joonho;Karouzos, Marios;Im, Myungshin
    • The Bulletin of The Korean Astronomical Society
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    • v.41 no.1
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    • pp.54.1-54.1
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    • 2016
  • Multi-wavelength variability is a staple of active galactic nuclei (AGN). Optical variability probes the nature of the central engine of AGN at smaller linear scales than conventional imaging and spectroscopic techniques. Previous studies have shown that optical variability is more prevalent at longer timescales and at shorter wavelengths. Intra-night variability can be explained through the damped random walk model but small samples and inhomogeneous data have made constraining this model hard. To understand the properties and physical mechanism of intra-night optical variability, we are performing the KMTNet Active Nuclei Variability Survey (KANVaS). Using KMTNet, we aim to study the intra-night variability of ~1000 AGN at a magnitude depth of ~19mag in R band over a total area of ${\sim}24deg^2$ on the sky. Test data in the COSMOS, XMM-LSS, and S82-2 fields was obtained over 4, 6, and 8 nights respectively during 2015, in B, V, R, and I bands. Each night was composed of 5-13 epoch with ~30 min cadence and 80-120 sec exposure times. As a pilot study, we analyzed data in the COSMOS field where we reach a magnitude depth of ~19.5 in R band (at S/N~100) with seeing varying between 1.5-2.0 arcsec. We used the Chandra-COSMOS catalog to identify 166 AGNs among 549 AGNs at B<23. We performed differential photometry between the selected AGN and nearby stars, achieving photometric uncertainty ~0.01mag. We employ various standard time-series analysis tools to identify variable AGN, including the chi-square test. Preliminarily results indicate that intra-night variability is found for ~17%, 17%, 8% and 7% of all X-ray selected AGN in the B, V, R, and I band, respectively. The majority of the identified variable AGN are classified as Type 1 AGN, with only a handful of Type 2 AGN showing evidence for variability. The work done so far confirms there are more variable AGN at shorter wavelengths and that intra-night variability most likely originates in the accretion disk of these objects. We will briefly discuss the quality of the data, challenges we encountered, solutions we employed for this work, and our updated future plans.

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