• Title/Summary/Keyword: Biomedical technology

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Establishment of a NanoBiT-Based Cytosolic Ca2+ Sensor by Optimizing Calmodulin-Binding Motif and Protein Expression Levels

  • Nguyen, Lan Phuong;Nguyen, Huong Thi;Yong, Hyo Jeong;Reyes-Alcaraz, Arfaxad;Lee, Yoo-Na;Park, Hee-Kyung;Na, Yun Hee;Lee, Cheol Soon;Ham, Byung-Joo;Seong, Jae Young;Hwang, Jong-Ik
    • Molecules and Cells
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    • v.43 no.11
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    • pp.909-920
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    • 2020
  • Cytosolic Ca2+ levels ([Ca2+]c) change dynamically in response to inducers, repressors, and physiological conditions, and aberrant [Ca2+]c concentration regulation is associated with cancer, heart failure, and diabetes. Therefore, [Ca2+]c is considered as a good indicator of physiological and pathological cellular responses, and is a crucial biomarker for drug discovery. A genetically encoded calcium indicator (GECI) was recently developed to measure [Ca2+]c in single cells and animal models. GECI have some advantages over chemically synthesized indicators, although they also have some drawbacks such as poor signal-to-noise ratio (SNR), low positive signal, delayed response, artifactual responses due to protein overexpression, and expensive detection equipment. Here, we developed an indicator based on interactions between Ca2+-loaded calmodulin and target proteins, and generated an innovative GECI sensor using split nano-luciferase (Nluc) fragments to detect changes in [Ca2+]c. Stimulation-dependent luciferase activities were optimized by combining large and small subunits of Nluc binary technology (NanoBiT, LgBiT:SmBiT) fusion proteins and regulating the receptor expression levels. We constructed the binary [Ca2+]c sensors using a multicistronic expression system in a single vector linked via the internal ribosome entry site (IRES), and examined the detection efficiencies. Promoter optimization studies indicated that promoter-dependent protein expression levels were crucial to optimize SNR and sensitivity. This novel [Ca2+]c assay has high SNR and sensitivity, is easy to use, suitable for high-throughput assays, and may be useful to detect [Ca2+]c in single cells and animal models.

10 MV X-ray Beam Dosimetry by Water and White Polystyrene Phantom (물과 백색폴리스티렌 팬텀에 의한 10 MV X-선 빔 선량계측)

  • Kim, Jong-Eon;Cha, Byung-Youl;Kang, Sang-Sik;Park, Ji-Koon;Sin, Jeong-Wook;Kim, So-Yeong;Jo, Seong-Ho;Son, Dae-Woong;Choi, Chi-Won;Park, Chang-Hee;Yoon, Chun-Sil;Lee, Jong-Duk;Park, Byung-Do
    • Journal of radiological science and technology
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    • v.31 no.1
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    • pp.83-87
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    • 2008
  • The purpose of this study is to get the correction factor to correct the measured values of the absolute absorbed dose proportional to the water equivalent depth. The measurement conditions in white polystyrene and water phantoms for 10MV X-ray beam are that the distance of source to center of ionization chamber is fixed at SAD 100 cm, the field sizes are $10{\times}10\;cm^2$, $20{\times}20\;cm^2$ and the depths are 2.3 cm, 5 cm, 10 cm, and 15 cm, respectively. The mean value of ionization was obtained by three times measurements in each field size and depths after delivering 100 MU from linear accelerator with output of 400 MU per min to the two phantoms. The correction factor and the percentage deviation in TPR were obtained below 0.97% and 0.53%, respectively. Therefore, we can get high accuracy by using the correction factor and the percentage deviation in TPR in measuring the absolute absorbed dose with the solid water equivalent phantom.

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Human Brain Pyridoxal-5'-phosphate Phosphatase: Production and Characterization of Monoclonal Antibodies

  • Kim, Dae-Won;Eum, Won-Sik;Choi, Hee-Soon;Kim, So-Young;An, Jae-Jin;Lee, Sun-Hwa;Sohn, Eun-Joung;Hwang, Seok-Il;Kwon, Oh-Shin;Kang, Tae-Cheon;Won, Moo-Ho;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Choi, Soo-Young
    • BMB Reports
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    • v.38 no.6
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    • pp.703-708
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    • 2005
  • We cloned and expressed human pyridoxal-5'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin $B_6$, in Escherichia coli using pET15b vector. Monoclonal antibodies (mAb) were generated against purified human brain PLP phosphatase in mice, and four antibodies recognizing different epitopes were obtained, one of which inhibited PLP phosphatase. The binding affinities of these four mAbs to PLP phosphatase, as determined using biosensor technology, showed that they had similar binding affinities. Using the anti-PLP phosphatase antibodies as probes, we investigated their cross-reactivities in various mammalian and human tissues and cell lines. The immunoreactive bands obtained on Western blots had molecular masses of ca. 33 kDa. Similarly fractionated extracts of several mammalian cell lines all produced a single band of molecular mass 33 kDa. We believe that these PLP phosphatase mAbs could be used as valuable immunodiagnostic reagents for the detection, identification, and characterization of various neurological diseases related to vitamin $B_6$ abnormalities.

Evaluation of a PCR-Reverse Blot Hybridization Assay to Identify Six Dermatophytes Predominant in the Republic of Korea

  • Jin, Hyunwoo;Kim, Hyunjung;Kim, Sunghyun;Choi, Yeonim;Bang, Hyeeun;Park, Sangjung;Wang, Hyeyoung;Lee, Jang-Ho;Jang, In Ho;Kim, Young-Kwon;Lee, Hyeyoung
    • Biomedical Science Letters
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    • v.20 no.3
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    • pp.139-146
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    • 2014
  • Accurate and rapid diagnosis of dermatophytosis, a disease whose prevalence has been steadily increased, is important for successful treatment. Current laboratory methods for diagnosing dermatophytosis rely on KOH mount and fungal culture method. However, these methods have low sensitivity and are time-consuming (2~4 weeks to diagnosis). In our previous study, a rapid molecular diagnostic assay (PCR-reverse blot hybridization assay, REBA) was developed to identify the following 6 main species of dermatophytes: Trichophyton rubrum, T. mentagrophytes, T. tonsurans, Microsporum canis, M. gypseum, and Epidermophyton floccosum. However, the REBA required more evaluation to validate its use in clinical examinations. The aim of the present study was to evaluate and validate the ability of the PCR-REBA to successfully identify dermatophytes in clinical isolates from dermatophytosis patients. Both conventional identification methods and the PCR-REBA were used to assess the presence of species of dermatophytes in 148 clinical isolates. The results of the two approaches were compared, and discrepancies between the two approaches were resolved by fungal ITS1 sequence analysis. T. rubrum was the most prevalent dermatophyte identified by conventional identification methods (118/148, 79.7%) and the PCR-REBA (131/148, 88.4%). The overall rate of consistency between conventional identification methods and the PCR-REBA was 79.0% (117/148 samples). Fungal ITS1 sequence analysis showed that PCR-REBA results were correct for 93.5% (29/31) of the discrepant samples. The PCR-REBA is rapid, sensitive, and highly specific compared with conventional identification methods. Thus, the PCR-REBA is a potentially useful tool for identifying dermatophytes in clinical settings.

Intrabed and Interbed Networks for Patient Monitoring (환자 모니터링을 위한 인트라베드 및 인터베드 통신망)

  • Park, Seung-Hun;Woo, Eung-Je;Kim, Kyung-Soo;Choi, Keun-Ho;Kim, Seung-Tae;Lee, Hee-Cheol;Seo, Jae-Joon;Kim, Hyung-Jin
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.05
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    • pp.285-289
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    • 1997
  • In this paper, we describe the intrabed and interbed network in a developed patient monitoring system. Intrabed network handles data communication among the main unit of a bedside monitor and parameter modules plugged in it. Interbed network deals with a higher level data communication among many bedside monitors, central stations, DB servers, and clinical workstations. Analyzing the data communication requirements in each stage of the system, we designed the intrabed network based upon RS-485 and HDLC protocol with 1Mbps data rate. Interbed network is designed to utilize the industry standard 10Base-T Ethernet with TCP/IP and UDP protocol. We present the specifications and the performances of the developed data communication networks in the patient monitoring system.

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Anti-inflammatory Properties of Meso-dihydroguaiaretic Acid in Lipopolysaccharide-induced Macrophage

  • Kim, Yong-Jae;Kang, Yeo-Jin;Kim, Tack-Joong
    • Biomedical Science Letters
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    • v.16 no.2
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    • pp.91-95
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    • 2010
  • Meso-dihydroguaiaretic acid (MDGA) is a medicinal herbal product isolated from the bark of Machilus thunbergii Sieb. et Zucc. (Lauraceae). It exhibits a neuroprotective effect and also exerts cytotoxicity to certain cancer cells. In the present study, we investigated whether or not MDGA inhibits inflammatory reaction through the inhibition of nitric oxide (NO) generation. The results showed that MDGA (5~$25 {\mu}M$) inhibited 100 ng/ml lipopolysaccharide (LPS)- induced NO generation in macrophage Raw 264.7 cells in a concentration-dependent manner. We also measured the cytotoxic effects of MDGA on Raw 264.7 cells and found no evidence of cytotoxicity. The inhibition of NO generation by MDGA was consistent with the inhibitory effect on the expression of inducible nitric oxide synthase (iNOS). In addition, MDGA inhibited the LPS-induced gene expression of $interleukin-1{\beta}$ $(IL-1{\beta})$ as well as tumor necrosis $factor-{\alpha}$ $(TNF-{\alpha})$. The present results may provide that MDGA has anti-inflammatory properties through inhibition of the toll-like receptors (TLRs) pathway, and suggest that MDGA can be used as an anti-inflammatory agent.

Position Recognition and User Identification System Using Signal Strength Map in Home Healthcare Based on Wireless Sensor Networks (WSNs) (무선 센서네트워크 기반 신호강도 맵을 이용한 재택형 위치인식 및 사용자 식별 시스템)

  • Yang, Yong-Ju;Lee, Jung-Hoon;Song, Sang-Ha;Yoon, Young-Ro
    • Journal of Biomedical Engineering Research
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    • v.28 no.4
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    • pp.494-502
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    • 2007
  • Ubiquitous location based services (u-LBS) will be interested to an important services. They can easily recognize object position at anytime, anywhere. At present, many researchers are making a study of the position recognition and tracking. This paper consists of postion recognition and user identification system. The position recognition is based on location under services (LBS) using a signal strength map, a database is previously made use of empirical measured received signal strength indicator (RSSI). The user identification system automatically controls instruments which is located in home. Moreover users are able to measures body signal freely. We implemented the multi-hop routing method using the Star-Mesh networks. Also, we use the sensor devices which are satisfied with the IEEE 802.15.4 specification. The used devices are the Nano-24 modules in Octacomm Co. Ltd. A RSSI is very important factor in position recognition analysis. It makes use of the way that decides position recognition and user identification in narrow indoor space. In experiments, we can analyze properties of the RSSI, draw the parameter about position recognition. The experimental result is that RSSI value is attenuated according to increasing distances. It also derives property of the radio frequency (RF) signal. Moreover, we express the monitoring program using the Microsoft C#. Finally, the proposed methods are expected to protect a sudden death and an accident in home.

Effects of Unloading induced Denervation on Trabecular Bone of Growing Mice; Gender-difference (궁둥신경절제술을 통한 무부하가 성별에 따른 성장기 쥐 해면골에 미치는 영향)

  • Seo, Dong-Hyun;Ko, Chang-Yong;Kang, Sun-Young;Lim, Do-Hyung;Kim, Han-Sung
    • Journal of the Korean Society for Precision Engineering
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    • v.26 no.10
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    • pp.122-128
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    • 2009
  • The aim of this study was to investigate, through a longitudinal tracking, whether gender influences morphological characteristics on trabeucular bone of tibia during unloading. 12 male and 15 female ICR mice at 6 week of age were used and randomly allocated into two groups in each gender; unloading and normal group, respectively Mice in unloading group were operated on denervation (sciatic neurectomy). The tibiae were scanned by using in vivo micro-computed tomography (micro-CT) at 0 day (before denervation) and after 14 days. Structural parameters and BMD on tibia were measured. In female, BV/TV (65%), Tb. Th (5%), Tb.N (61%) and BMD (62%) were significantly decreased and BS/BV (17%), Tb.Sp (54%), SMI (19%) and Tb.Pf (75%) were significantly increased after unloading (p<0.05). In male, BV/TV (1%) and Tb.N (10%) were significantly decreased and SMI (5%) was significantly increased after unloading (p<0.05). In addition, trabeuclar bone loss of female was significantly bigger than that of male (p<0.05). These results indicated that effects of unloading on trabecular bone in growing mice might have difference between female and male, although unloading result in loss of quantity and quality on trabecular bone both female and male.

Demonstration of the Usefulness of Optical Coherence Tomography in Imaging a Mouse Tail Model of Lymphedema

  • Kim, Hui Dong;Kim, Dong Kyu;Chae, Yu-Gyeong;Park, Seok Gyo;Kim, Ghi Chan;Jeong, Ho Joong;Sim, Young-Joo;Ahn, Yeh-Chan
    • Current Optics and Photonics
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    • v.1 no.2
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    • pp.132-137
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    • 2017
  • To investigate the usefulness of optical coherence tomography (OCT) for imaging lymphedema, we directly compared it to other histological methods in a mouse model of lymphedema. We performed detailed imaging of the lymphedema lesion on a mouse tail. We imaged the mouse tail in vivo with OCT and created histopathological samples. We constructed a spectrometer-based OCT system using a fiber-optic Michelson interferometer. The light was directed to 50:50 couplers that split the light into reference and sample arms. Backscattered light from a reference mirror and the sample produced an interference fringe. An OCT image of the lymphedema model revealed an inflammatory reaction of the skin that was accompanied by edema, leading to an increase in the light attenuation in the dermal and subcutaneous layers. Similar to OCT image findings, histological biopsy showed an inflammatory response that involved edema, increased neutrophils in epidermis and subdermis, and lymphatic microvascular dilatation. Furthermore, the lymphedema model showed an increase in thickness of the dermis in both diagnostic studies. In the mouse tail model of lymphedema, OCT imaging showed very similar results to other histological examinations. OCT provides a quick and useful diagnostic imaging technique for lymphedema and is a valuable addition or complement to other noninvasive imaging tools.

Analysis of Eddy Current Effect in Magnetic Resonance Imaging Using the Finite Element Method (유한요소법에 의한 자기공명영상시스템에서의 와전류 영향 분석)

  • Lee, Jeong-Han;Gang, Hyeon-Su;Jo, Min-Hyeong;Mun, Chi-Ung;Lee, Gang-Seok;Lee, Su-Yeol
    • Journal of Biomedical Engineering Research
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    • v.20 no.1
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    • pp.53-58
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    • 1999
  • Eddy current in MRI systems degrades gradient field linearity and distorts gradient waveform. When the waveform distortion is spatially variant, it is very difficult to perform special imaging techniques such as the echo planar imaging technique or the fast spin echo imaging technique. In this study, we have developed a new technique to estimate the distorted gradient waveforms at any points inside the imaging region using the finite element method. After obtaining the eddy-current-effect transfer function, which represents magnitude and phase characteristics of the gradient field at a particular point, we have used the transfer function to estimate the actual gradient waveforms at the point. To verify the proposed technique, we have compared the estimated gradient waveforms with the measured ones.

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