• Title/Summary/Keyword: 염색 기법

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Development of a New Artificial Latent Fingerprint Aqueous Solution by Improving Lipid Composition (지질조성 개선을 통한 새로운 인공 잠재지문 수용액의 개발)

  • Sang-Yoon LEE;Hwa-Seon LIM;Ki-Jong RHEE
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.1
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    • pp.99-103
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    • 2024
  • Previous artificial latent fingerprint solution has shown unsatisfactory results. Therefore, in this study, we developed an artificial latent fingerprint solution close to the actual fingerprint composition by improving the lipid composition. We mixed lipid solution with amino acid solution at v/v ratios as follows: 2:3, 1:4, 1:5, 1:8, 1:10, 1:20. We then dropped the same amount of each proportion of artificial latent fingerprint solution on porous paper and non-porous slide glass. Subsequently, each sample was treated with Oil red O, Cyanoacrylate fuming and Basic yellow 40 staining. As the concentration of lipids decreased, the output also decreased. Both types of surfaces and all concentrations were visually confirmed very well. In addition, the reactivity to lipids was significantly higher compared to the previous artificial latent fingerprint solution. Furthermore, for the quantitative evaluation, it is necessary to conduct additional research on the printing of the artificial latent fingerprint solution.

Aberrant Microtubule Assembly and Chromatin Configuration of Homan Oocytes Which Failed to Complete Fertilization Following In Vitro Fertilization and Intracytoplasmic Sperm Injection (일반적 수정과 세포질내 정자주입법에 의해 수정에 실패한 인간난자의 미세소관과 염색체의 형태이상)

  • Chung, H. M.;Kim, N. H.;Kim, J. W.;J. M. Lim;Park, C.;J. J. Ko;K. Y. Cha;Kim, J. M.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.2
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    • pp.143-154
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    • 2000
  • Most eggs initiated the fertilization processes but arrested at specific stages. The stages included failure of the oocyte to exit from the meiotic metaphase-II with or without sperm penetration, failure of appropriate sperm aster formation, inability to form proper male and female pronuclei, failure of suitable pronuclear apposition, and failure to form proper number of either male or female pronuclei. Various images of defective microtubule organization and chromatin configuration during IVF and ICSI procedures were observed. We discussed the data with previous research results during normal fertilization in humans and other mammals. In conclusion, various aberrant patterns in microtubule assembly and chromatin configuration, which were assessed in the present study, could be used as criteria to improve assisted reproductive technology in clinics. However, further cellular and molecular characterization is needed to clarify these aberrant patterns of cytoskeletal assembly.

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Application of Molecular Diagnostics Technology in the Development of a Companion Diagnostics for Malignant Solid Tumors (악성 고형암의 항암제 동반진단 기술에서 분자진단기술의 적용)

  • Kim, Jin-Hee
    • The Journal of the Korea Contents Association
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    • v.19 no.3
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    • pp.365-374
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    • 2019
  • Unlike benign tumors, malignant tumors are capable of metastasis, easy to relapse, poor survival, and low quality of life. In Korea, here is a tendency to treat the tumors collectively according to the General Principles of Cancer Chemotherapy(GPCC) of the Health Insurance Review & Assessment Service (HIRA). But recently, companion diagnostics(CDx) is recommended rather than unilateral medication because biomarker-based molecular diagnostics is possible to predict the drug response of patients before drug treatment. Not only domestic but also overseas Food and Drug Administratio (FDA) recommends the development of the CDx system at the stage of drug development to ensure the responsiveness and safety of medicines. In this study, I focused on the necessity of CDx development direction as well as CDx development status through literature review. Furthermore I also discussed CDx types according to the molecular diagnostic technology such as immunohistochemistry (IHC), polymerase chain reaction (PCR), in situ hybridization (ISH), and next-generation sequencing (NGS) not only in the approved CDx but also in the developing one by US FDA. And I suggested the technology issue of CDx development process such as a selection of molecular diagnostics at the time of release, a clear understanding of the CDx mechanism, and a convergence of drug with CDx development. The necessity of social insurance system also was proposed for CDx development.

Application of Chemiluminescence Enzyme Immunoassay Method to Collect in vivo Matured Oocyte in Dog Cloning (개 복제 시 체내 성숙 난자 회수를 위한 화학발광효소면역분석기법의 적용)

  • Kim, Min-Jung;Oh, Hyun-Ju;Kim, Geon-A;Jo, Young-Kwang;Choi, Jin;Lee, Byeong-Chun
    • Journal of Veterinary Clinics
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    • v.31 no.4
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    • pp.267-271
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    • 2014
  • Accurate determination of in vivo oocyte maturation is particularly critical for dog cloning compared to other assisted reproductive technologies because oocytes in metaphase II stage have to be recovered in order to undergo somatic cell nuclear transfer right after its recovery. The aim of present study was to evaluate the reliability and to set a reference range of a chemiluminescence enzyme immunoassay (CLEIA) compared to radioimmunoassay (RIA) method to retrieve in vivo matured oocytes. Serum progesterone concentration during proestrus and estrus was analyzed by RIA and CLEIA to determine ovulation day (Day 0). On Day 3, in vivo oocytes were recovered surgically and evaluated microscopically maturation status after staining nucleus with bisbenzimidazole dye. Mean progesterone concentration by CLEIA ($7.64{\pm}0.06ng/ml$) was significantly higher than by RIA ($6.46{\pm}0.04ng/ml$, P < 0.0001). It was not different between CLEIA ($10.01{\pm}0.34ng/ml$) and RIA values ($7.91{\pm}0.14ng/ml$, P < 0.05) on Day 0, but significantly higher CLEIA level on Day -1 and Day 1 ($6.41{\pm}0.15$ and $14.25{\pm}0.44ng/ml$) was assessed compared to RIA ($4.95{\pm}0.10$ and $11.29{\pm}0.34ng/ml$). However, with both methods, progesterone level was significantly increased from Day -1 to Day 2. To determine oocyte maturation with CLEIA method, a wider and higher reference range has to be considered.

Human Blastocysts;The Correlation Between Embryo Microscopical Assessments and Their Cell Number (인간 배반포기 배의 현미경적 분류와 세포수의 상관관계에 관한 연구)

  • Kim, E.Y.;Uhm, S.J.;Kim, M.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.319-326
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    • 1996
  • The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.

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Biological Activity of Mixed Extracts of Acanthopanax senticosus and Citrus unshiu Fermented with Bovista plumbea for Inner Beauty (이너뷰티 소재로서의 생물전환된 가시오가피-진피 혼합 추출물의 생물학적 활성)

  • Eun Jeong Kim;So Yeon Kim;Su Yeon Kang;Yung Choon Yoo;Taek Joon Yoon;Gye Won Lee;Young Ho Cho
    • Journal of Life Science
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    • v.33 no.7
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    • pp.555-564
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    • 2023
  • In this study, the anti-oxidant activity, elastase inhibitory activity, and skin moisturizing effect of mixed extracts of Acanthopanax senticosus and Citrus unshiu fermented with Bovista plumbea (B-MEAC) were evaluated to verify the availability as a material for inner beauty. The DPPH radical scavenging activity of B-MEAC was showed in a dose-dependent manner (SC50=156.1±0.82 ㎍/ml). Also, B-MEAC inhibited the elastase activity in a concentration-dependent manner (p<0.001). To study the effect of B-MEAC on mouse skin hydration, skin moisture content and transepidermal water loss (TEWL) measured. As a result, skin moisture content increased (p<0.001) and TEWL decreased (p<0.01) compared to the dry-induced control group. The effect on the change of collagen fibers in the dry-induced mouse skin was examined through Masson's trichrome staining. In the group administered with B-MEAC, the amount of collagen relatively increased compared to the control group, and the intensity of blue color increased. The effect on the moisturizing function of the dry-induced mouse skin was examined by Western blot method. In the group administered with B-MEAC, the expression of matrix metalloproteinase-1 (MMP-1) protein decreased compared to the control group. In addition, the expression level of collagen1A1 (COL1A1), hyaluronan synthase-2 (HAS2), filaggrin, and aquaporin-3 (AQP3) recovered (p<0.001). Therefore, these results suggest the potential of B-MEAC as a skin hydration agent for inner beauty.

Development of Proteomics-based Biomarkers for 4 Korean Cultivars of Sorghum Seeds (Sorghum bicolor (L.) Moench) (국내 수수 종자 분석을 위한 프로테오믹스-기반 바이오마커 개발)

  • Kim, Jin Yeong;Lee, Su Ji;Ha, Tae Joung;Park, Ki Do;Lee, Byung Won;Kim, Sang Gon;Kim, Yong Chul;Choi, In Soo;Kim, Sun Tae
    • Korean Journal of Environmental Agriculture
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    • v.32 no.1
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    • pp.48-54
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    • 2013
  • BACKGROUND: Sorghum (Sorghum bicolor (L.) Moench) ranks as the 6th most planted crop in the world behind wheat, rice, maize, soybean, and barley. The objective of this study was to identify bio-marker among sorghum cultivars using proteomics approach such as two-dimensional polyacrylamide gel electrophoresis (2-DE) coupled with mass spectrometry (MS). METHODS AND RESULTS: Proteins were extracted from sorghum seed, and separated by 2-DE. Total 652 spots were detected from 4 different sorghum seed after staining of 2-DE with colloidal Coomassie brilliant blue (CBB). Among them, 8 spots were differentially expressed and were identified using MALDI-TOF/TOF mass spectrometry. They were involved in RNA metabolism (spot1, spot 4), heat shock proteins (HSPs, spot 2), storage proteins (spot 3, spot 5, and spot 6), and redox related proteins (spot 8). Eight of these proteins were highly up-regulated in Whinchalsusu (WCS). The HSPs, Cupin family protein, and Globulin were specifically accumulated in WCS. The DEAD-box helicase was expressed in 3 cultivars except for WCS. Ribonuclease T2 and aldo-keto reductase were only expressed in 3 cultivars except for Daepung-susu (DPS). CONCLUSION(S): Functions of identified proteins were mainly involved in RNA metabolism, heat shock protein (HSP), and redox related protein. Thus, they may provide new insight into a better understanding of the charactreization between the cultivars of sorghum.

A Study on the Localization of Neurofascin in the Myelinated Rat Sciatic Nerve Fibers (랫드 수초좌골신경섬유에서 Neurofascin분포에 대한 면역세포화학적 연구)

  • Chang, Byung-Hwa;You, Kwan-Hee;Lee, Jong-Hwan;Cho, Ik-Hyun;Bae, Chun-Sik;Park, Chang-Hyun;Han, Jeong-Mi;Choe, Nong-Hoon;Chang, Byung-Joon
    • Applied Microscopy
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    • v.36 no.2
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    • pp.131-140
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    • 2006
  • Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the myelination of peripheral nerves, the localization of neurofascin in myelinated rat sciatic nerve fibers was studied with the immuno-fluorescence and immune-electron microscopy and the results are as follows; 1. According to the myelination is going on, neurofascin localization was dramatically changed in the sciatic nerve fibers. 2. In the myelinated fibers, neurofascin was weakly localized along the axolemma at the node of Ranvier. 3. Neurofascin was also apparantly localized at the non-compact area of Schwann cell membrane such as paranodal loop, Schmidt-Lantermann incisure, inner & outer mesaxons in the myelinated fibers. From the above results, neurofascin is likely to have a role to sustain the ideal gap of apposing membranes of Schwann cell, so it may enable to materials transport in the myelin sheath.

PHOTOTOXIC EFFECTS OF LOW LEVEL LASER IRRADIATION ON HUMAN OSTEOSARCOMA CELLS (골육종세포에 미치는 레이저 조사의 광독성 효과)

  • Son, Jang-Ho;Cho, Young-Chul;Ryu, Sung-Ho;Kim, Gyoo-Cheon;Sung, Iel-Yong;Park, Bong-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.6
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    • pp.509-515
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    • 2004
  • Photodynamic therapy(PDT) has advanced to clinical trials for the treatment of a variety of solid tumors and presents an alternative treatment option for tumors resistant to chemo-and/or radio-therapy. PDT is based on the combination of laser light of appropriate wavelength and energy to activate a systemically or locally applied photosensitizer that concentrates preferentially in malignant tissues. In this study, phototoxicity of laser EIT 21 was analysed in human osteosarcoma cell(HOS) and the second objective of this study was to determine the ability of laser EIT 21 to induce apoptosis. This study demonstrated that laser EIT 21 had a phototoxicity to HOS cells. In order to examinate whether cell death was induced by necrosis or apoptosis, variety of techniques which assess apoptosis were used. TUNEL assay showed only a few the positive reaction on condensed nuclei. It is hard to find condensed or fragmented nuclei on HOS cells irradiated with laser EIT 21 in Hemastat and AO/EB stain. By DNA electrophoresis, cells also did not show DNA degradation characteristic of apoptosis with a ladder pattern of DNA fragments. Apoptosis-related factors were analyzed by western blotting. The expression of p53 was constant and cells irradiated with laser did not show the caspase-3 and PARP degradation, therefore we suggest that p53 and caspase-3 are not involved in laserinduced cell death.

Cultivation and Electron Microscopy of Bovine Leukemia Virus from Peripheral Blood Lymphocytes of Holstein-Friesian Dairy Cattle (젖소 말초혈액 림프구로부터 소백혈병 바이러스 배양 및 전자현미경적 관찰)

  • Yoon, Soon-Seek;Park, Jung-Won;Byun, Jae-Won;Kang, Mun-Il;Yoo, Han-Sang;Han, Hong-Ryul
    • Applied Microscopy
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    • v.35 no.1
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    • pp.23-30
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    • 2005
  • Many studies have been performed on the bovine leukemia virus (BLV) since bovine leukosis had been reported in 1968 in Korea. However, there was no report on the ultrastructural examination of BLV. An attempt to detect C-type viral particles in the cultured peripheral blood lymphocytes of Holstein-Friesian dairy cattle, was made to determine whether in vitro viral expression might be used as a reliable method to identify the cow which is likely to transmit BLV. In transmission electron microscopic (TEM) examination, the virus particles were found predominantly outside of the lymphocytes even though a few particles were also observed within the membrane bound cytoplasmic vacuoles. All of them were C-type particles consisting of a central, electron-dense core separated by a clear area from a limiting envelope with a unit membrane structure. Virus particles were easily detected in the lymphocyte which was cultured with medium supplemented with either T-lymphocyte mitogen (conconavalin A) or B-lymphocyte mitogen (lipopolysaccharide). Identical viral particles, although fewer, were also consistently present in the lymphocytes cultured with medium which was containing foetal bovine serum (FBS) only and which was containing neither FBS or mitogen. By contrast, no virus particle was detected in extensive examination of lymphocytes before culture. In conclusion, the BLV cultivation and detection methods established in this study could be used as a tool to identify and eliminate the cattle which can transmit the BLV.