• Title/Summary/Keyword: In-vitro diagnosis

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Tumor Uptake Rate of Alkaline $^{99m}Tc-DMSA$ in Walker Carcinosarcoma 256 Bearing Wistar Rats (염기성 $^{99m}Tc-DMSA$의 Walker 256 암육종 이식백서에서 종양섭취율에 관한 연구)

  • Lim, S.M.;Hong, S.W.;Awh, O.D.;Lee, M.C.;Koh, C.S.
    • The Korean Journal of Nuclear Medicine
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    • v.22 no.1
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    • pp.65-76
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    • 1988
  • High affinity complexes for the tumor were obtained by changing pH and composition in the preparation of $^{99m}Tc-DMSA$. The purpose of this study was to investigate the tumor affinity, and in vitro and in vivo characteristics of these complexes. The results obtained were as follows; 1) Tumor imaging agent was formed successfully at pH $6.0\sim9.0$ and renal imaging agent at pH $2.0\sim5.0$. 2) The serum protein binding of $^{99m}Tc-DMSA$ was $89.1\sim92.8%$ at pH $2.0\sim5.0$ and $11.8\sim30.5%$ at pH $6.0\sim9.0$ respectively, and it was not changed with time. 3) The T 1/2 of tumor affinity complex in blood between 3 and 6 hours after injection was $187{\pm}29$ minutes $(mean{\pm}SD)$. 4) In the blood, the radioactivity was mainly in the plasma, and less than 1% was in the cellular components. 5) In the Walker carcinosarcoma 256 bearing Wistar rats, the radioactivity in the kidney increased, and decreased in the skeleton with time. The radioactivity in the tumor showed the peak in 6 hours after injection and decreased thereafter. 6) In the tumor cell, the radioactivity localized mainly in the cytosol, the soluble fraction of the cytoplasm. This study provides the basic knowledge about tumor affinity and usefulness of $^{99m}Tc-DMSA$ in the diagnosis of malignant disease.

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An Usefulness of In Vitro Interferon Gamma Assay for the Diagnosis of Latent Tuberculosis Infection in Middle- and High-School Students in Jeju-Shi, Korea (제주시 중고등학생의 잠복결핵감염 진단을 위한 체외 인터페론감마검사의 유용성)

  • Bae, Jong-Myon;Kim, Eun-Hee;Wang, Ok-Bo
    • Tuberculosis and Respiratory Diseases
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    • v.68 no.3
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    • pp.155-161
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    • 2010
  • Background: The tuberculin skin test (TST) has limitations in diagnosing a latent tuberculosis infection (LTBI). The interferon-gamma release assay (IGRA) was introduced to middle- and high-school students since 2009 by the Korea Centers for Disease Control and Prevention. The aim was to evaluate the utility of IGRA in diagnosing LTBI in middle- and high-school students. Methods: From August 2007 to July 2009, among suspected LTBI students showing TST induration with a 10 mm diameter and over with a normal chest x-ray in school students of Jeju city, 341 students underwent a Quanti FERON-TB Gold In-Tube (QFT-IT) test to confirm LTBI. Results: From 348 students showing a positive TST, a QFT-IT test was carried out on 341 students. The positive QFT-IT rate was 52.8% (=180/341). The positive QFT-IT rate was higher in high-school boys with a 15~19 mm diameter of induration in TST. Conclusion: With the introduction of IGRA for diagnosing LTBI in middle- and high-school students, approximately 47% of students who show a TST induration with a 10 mm diameter and over can avoid taking unnecessary preventive chemotherapy. These results suggest that IGRA is useful for diagnosing and controlling LTBI in Korean students.

Detection and Epidemiological Survey of Canine Parvoviral Enteritis by Polymerase Chain Reaction (Polymerase Chain Reaction을 이용한 Canine Parvovirus성장염의 진단과 역학조사)

  • Kim, Doo;Jang, Wook
    • Journal of Veterinary Clinics
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    • v.14 no.2
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    • pp.177-184
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    • 1997
  • Canine parvovirus(CPV) is a very highly contagious virus causing hemorrhagic enteritis and myocarditis mainly in young dogs. The diseases were first recognized in 1978, and then spread throughout the world by 1980. The main source of the infection seems to be the feces of infected dogs, at the same time feces are suitable materials for detection of virus in the enteric form exactly for the same reasons. Recently, a new technique of in vitro DNA amplification, Known as the polymerase chain reaction (PCR), has been widely applied to clinical viral diagnosis because of its sensitivity, specificity and rapidity. In this research, we attemped to set up the PCR for the detection of CPV in fecal samples and conformed the canine parvpviral enteritis by PCR. To increase the sensitivity and specificity of a PCR, the nested PCR (two-step PCR) was performed. We also surveyed the contamination status of CPV in the research using fecal specimen was highly sensitive and specific. Of the 100 fecal specimens suspected canine parvoviral enteritis, 45 fecal specimens were positive in HA test, 64 fecal specimens were positive in the first PCR, and 87 fecal specimens were positive in the second PCR. CPV contamination status of animal clinics and breeding centers was serious, wo hygienic management of environment in which dogs are reared is required. The nested PCR described here seems to be a rapid, sensitive and specific for the detection of canine parvovirus.

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Effect of object position in the field of view and application of a metal artifact reduction algorithm on the detection of vertical root fractures on cone-beam computed tomography scans: An in vitro study

  • Nikbin, Ava;Kajan, Zahra Dalili;Taramsari, Mehran;Khosravifard, Negar
    • Imaging Science in Dentistry
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    • v.48 no.4
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    • pp.245-254
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    • 2018
  • Purpose: To assess the effects of object position in the field of view (FOV) and application of a metal artifact reduction (MAR) algorithm on the diagnostic accuracy of cone-beam computed tomography (CBCT) for the detection of vertical root fractures(VRFs). Materials and Methods: Sixty human single-canal premolars received root canal treatment. VRFs were induced in 30 endodontically treated teeth. The teeth were then divided into 4 groups, with 2 groups receiving metal posts and the remaining 2 only having an empty post space. The roots from different groups were mounted in a phantom made of cow rib bone, and CBCT scans were obtained for the 4 different groups. Three observers evaluated the images independently. Results: The highest frequency of correct diagnoses of VRFs was obtained with the object positioned centrally in the FOV, using the MAR algorithm. Peripheral positioning of the object without the MAR algorithm yielded the highest sensitivity for the first observer (66.7%). For the second and third observers, a central position improved sensitivity, with or without the MAR algorithm. In the presence of metal posts, central positioning of the object in the FOV significantly increased the diagnostic sensitivity and accuracy compared to peripheral positioning. Conclusion: Diagnostic accuracy was higher with central positioning than with peripheral positioning, irrespective of whether the MAR algorithm was applied. However, the effect of the MAR algorithm was more significant with central positioning than with peripheral positioning of the object in the FOV. The clinical experience and expertise of the observers may serve as a confounder in this respect.

High-Level Production of High-Purity Human and Murine Recombinant Prion Proteins Functionally Compatible to In Vitro Seeding Assay

  • Hwang, Hae-Gwang;Kim, Dae-Hwan;Lee, Jeongmin;Mo, Youngwon;Lee, Se-Hoon;Lee, Yongjin;Hyeon, Jae Wook;Lee, Sol Moe;Cheon, Yong-Pil;Choi, Eun-Kyoung;Kim, Su Yeon;Lee, Yeong Seon;Son, Young-Jin;Ryou, Chongsuk
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1749-1759
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    • 2018
  • Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

Clinical application of genome-wide single nucleotide polymorphism genotyping and karyomapping for preimplantation genetic testing of Charcot-Marie-Tooth disease

  • Kim, Min Jee;Park, Sun Ok;Hong, Ye Seul;Park, Eun A;Lee, Yu Bin;Choi, Byung-Ok;Lee, Kyung-Ah;Yu, Eun Jeong;Kang, Inn Soo
    • Journal of Genetic Medicine
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    • v.19 no.1
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    • pp.7-13
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    • 2022
  • Purpose: Preimplantation genetic testing for monogenic disorders (PGT-M) has been successfully used to prevent couples with monogenic disorders from passing them on to their child. Charcot-Marie-Tooth Disease (CMT) is a genetic disorder characterized by progressive extremity muscle degeneration and loss of sensory function. For the first time in Korea, we report our experience of applying single nucleotide polymorphism genotyping and karyomapping for PGT-M of CMT disease. Materials and Methods: Prior to clinical PGT-M, preclinical tests were performed using genotypes of affected families to identify informative single-nucleotide polymorphisms associated with mutant alleles. We performed five cycles of in vitro fertilization PGT-M in four couples with CMT1A, CMT2A, and CMT2S in CHA Fertility Center, Seoul Station. Results: From July 2020 through August 2021, five cycles of PGT-M with karyomapping in four cases with CMT1 and CMT2 were analyzed retrospectively. A total of 17 blastocysts were biopsied and 15 embryos were successfully diagnosed (88.2%). Ten out of 15 embryos were diagnosed as unaffected (66.7%). Five cycles of PGT-M resulted in four transfer cycles, in which four embryos were transferred. Three clinical pregnancies were achieved (75%) and the prenatal diagnosis by amniocentesis for all three women confirmed PGT-M of karyomapping. One woman delivered a healthy baby uneventfully and two pregnancies are currently ongoing. Conclusion: This is the first report in Korea on the application of karyomapping in PGT-M for CMT patients. This study shows that karyomapping is an efficient, reliable and accurate diagnostic method for PGT-M in various types of CMT diseases.

Circular RNA hsa_circ_0075828 promotes bladder cancer cell proliferation through activation of CREB1

  • Zhuang, Chengle;Huang, Xinbo;Yu, Jing;Gui, Yaoting
    • BMB Reports
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    • v.53 no.2
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    • pp.82-87
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    • 2020
  • Circular RNAs (circRNAs), one kind of non-coding RNA, have been reported as critical regulators for modulating gene expression in cancer. In this study, microarray analysis was used to screen circRNA expression profiles of bladder cancer (BC) 5637 cells, T24 cells and normal control SV-HUC-1 cells. The data from the microarray showed that hsa_circ_0075828 (named circCASC15) was most highly expressed in 5637 and T24 cells. circCASC15 was highly expressed in BC tissues and cells. Overexpression of circCASC15 was closely associated with BC tumor stage and promoted cell proliferation significantly in vitro and in vivo. Mechanistically, circCASC15 could act as miR-1224-5p sponge to activate the expression of CREB1 to promote cell proliferation in BC. In short, circCASC15 promotes cell proliferation in BC, which might be a new molecular target for BC diagnosis and therapy.

Scabraside D Extracted from Holothuria scabra Induces Apoptosis and Inhibits Growth of Human Cholangiocarcinoma Xenografts in Mice

  • Assawasuparerk, Kanjana;Vanichviriyakit, Rapeepun;Chotwiwatthanakun, Charoonroj;Nobsathian, Saksit;Rawangchue, Thanakorn;Wittayachumnankul, Boonsirm
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.511-517
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    • 2016
  • Scabraside D, a sulfated triterpene glycoside extract from sea cucumber Holothulia scabra, shows various biological activities, but effects on human cholangiocarcinoma cells have not previously been reported. In the present study, we investigated the activity of scabraside D against human cholangiocarcinoma (HuCCA) both in vitro and for tumor growth inhibition in vivo using a xenograft model in nude mice. Scabraside D ($12.5-100{\mu}g/mL$) significantly decreased the viability and the migration of the HuCCA cells in a dose-dependent manner, with 50% inhibitory concentration (IC50) of $12.8{\pm}0.05{\mu}g/mL$ at 24 h. It induced signs of apoptotic cells, including shrinkage, pyknosis and karyorrhetic nuclei and DNA fragmentation on agarose gel electrophoresis. Moreover, by quantitative real-time PCR, scabraside D effectively decreased Bcl-2 while increasing Bax and Caspase-3 gene expression levels suggesting that the scabraside D could induce apoptosis in HuCCA cells. In vivo study demonstrated that scabraside D (1 mg/kg/day, i.p. for 21 days) significantly reduced growth of the HuCCA xenografts without adverse effects on the nude mice. Conclusively, scabraside D induced apoptosis in HuCCA cells and reduced the growth of HuCCA xenographs model. Therefore, scabraside D may have potential as a new therapeutic agent for cholangiocarcinoma.

Study on the Dependence of Ultrasonic Phase Velocity on Porosity, Frequency and Propagation Angle in Cancellous Bone (해면질골에서 다공율, 주파수 및 전파각에 대한 초음파 위상속도의 의존성 연구)

  • Lee, Kang-Il;Kim, Yong-Tae;Choi, Min-Joo
    • Journal of the Korean Society for Nondestructive Testing
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    • v.28 no.2
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    • pp.112-118
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    • 2008
  • In the present study, the dependence of ultrasonic phase velocity on porosity and frequency in cancellous bone was predicted using the Biot model and the modified Biot-Attenborough (MBA) model for propagation in fluid-saturated porous media. It was also compared with previously published measurements in human and bovine cancellous bones in vitro. It was shown that the phase velocity in cancellous bone decreased with increasing porosity and frequency The dependence of phase velocity on propagation angle in cancellous bone as predicted using the Schoenberg model together with the Biot model and tile MBA model which were modified to include the effect of angle. The theoretical models used in the present study advance our understanding of the interaction between ultrasound and cancellous bone and can be expected to be usefully employed for the diagnosis of osteoporosis.

Synthesis and Evaluation of Oleanolic Acid-Conjugated Lactoferrin for β-Amyloid Plaque Imaging

  • Kim, Sung-Min;Kim, Dongkyu;Chae, Min Kyung;Jeong, Il-Ha;Cho, Jee-Hyun;Choi, Naeun;Lee, Kyo Chul;Lee, Chulhyun;Ryu, Eun Kyoung
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3671-3675
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    • 2012
  • ${\beta}$-Amyloid accumulation in the brain is a pathological hallmark of Alzheimer's disease (AD). Since early detection of ${\beta}$-amyloid may facilitate more successful and timely therapeutic interventions, many investigators have focused on developing AD diagnostic reagents that can penetrate the blood-brain barrier (BBB). Oleanolic acid (OA) is a substance found in a variety of plants that has been reported to prevent the progression of AD in mice. In this study, we synthesized and evaluated a new radioligand in which OA was conjugated to lactoferrin (Lf, an iron-binding glycoprotein that crosses the BBB) for the diagnosis of AD. In an in vitro study in which OA-Lf was incubated with ${\beta}$-amyloid (1-42) aggregates for 24 h, we found that OA-Lf effectively inhibited ${\beta}$-amyloid aggregation and fibril formation. In vivo studies demonstrated that $^{123}I$-OA-Lf brain uptake was higher than$^{123}I$-Lf uptake. Therefore, radiolabeled OA-Lf may have diagnostic potential for ${\beta}$-amyloid imaging.