• Title/Summary/Keyword: phase contrast microscopy

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In vitro Cytotoxic Effect of Extracts from Styela plicata (오만둥이(Styela plicata) 추출물의 in vitro 세포독성 효과)

  • Lee, Bo-Bae;Cha, Mi-Ran;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.9
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    • pp.1099-1105
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    • 2007
  • The present study describes the preliminary evaluation of the anticancer activity of Styela plicata. Freeze-dried S. plicata was extracted with methanol, ethanol, acetone, and water, and then anticancer effect of the extracts was measured by the MTT reduction assay and phase-contrast microscopy on the HT-29 human colon carcinoma cells. Among the extracts, acetone extract showed the highest anticancer activity. The cell proliferation rates markedly decreased by 94.0% at the concentration of 500 ${\mu}g/mL$ of acetone extract compared with control cells. The acetone extract was further fractionated with hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The HT-29 cells with hexane layer extract (250 ${\mu}g/mL$) decreased the cell viability to 5.1% of untreated control. The growth of SW620, HeLa, and MCF-7 cells was decreased to about 10%, by the treatment of hexane layer extract 250 ${\mu}g/mL$. Theses results suggest extracts from S. plicata as possible natural cancer therapeutic material.

Analysis of dental hygiene assessment data of recall patients (mainly 20s age)

  • Choi, Hye-Jung;Woo, Hee-Sun
    • Journal of the Korea Society of Computer and Information
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    • v.27 no.6
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    • pp.131-137
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    • 2022
  • As the age increases, the oral cavity, that is, the teeth and periodontium, also begin to age, and accordingly, a preparation process is required. The preparation process is an important period for oral health management to start continuously with oral health education consisting of knowledge, attitude, and behavior from the 20s. Therefore, to design a clinical dental hygiene course for patients who visited a dental clinic in Gyeonggi-do and received continuous care in an oral health care room after treatment, we tried to analyze the data of the dental hygiene assessment. As a dental hygiene assessment tool, based on personal information and general medical history, dental visit experience, bleeding on probing(BOP), bad breath measurement, phase contrast microscopy, and O'Leary index were performed. The number of subjects who had dental visits was 75.4% and those without experience were 24.6%, and as a result of the periodontal examination, generally bleeding was found in 76.3%. In preventive oral care, the stage of dental hygiene assessment in the 20s is an important first step. From this point on, it is an important time to be systematically habituated so that you can take responsibility for your own oral condition. Therefore, in this study, the results of dental hygiene assessment through oral examinations of subjects in their 20s are derived and presented as basic data for the development of dental hygiene performance competency of dental hygienists during the clinical dental hygiene process in oral health education and oral health management.

Improvement of Reproductive Efficiency of Artificial Insemination Following Estrus Induction in Dog II. Semen Freezing and Artificial Insemination in Dog

  • Lee, Y. R.;S. Y. Choe
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.92-92
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    • 2002
  • Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to evaluate semen quality after cryopreservation and to evaluate the Pregnancy rate after insemination (AI). Fifty infertilie dogs (age 2∼3 years) were selected for the study and divided into three different estrus induction treatment groups. Group 1: dogs (n=15) were given clomifene (0.1 mg/kg) orally for five days at 12 hr intervals. Group 2: dogs (n=15) were given bromocriptine (50 $\mu\textrm{g}$/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Group 3, n=20) when pro-estrus occurred. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. The ejaculated semen to freeze was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM: TES, 209 mM: citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 cm above the surface of liquid nitrogen (LN$_2$) for 23 min. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, the pregnancy rate of dogs treated with a combination of GnRH and bromocriptine was more effective than use of clomifene or bromocriptine only. In addition, frozen-thawed semen can be used successfully far artificial insemination in dog.

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A Study on Characteristics of Airborne Asbestos Concentrations Using PCM and TEM in Life Environment Surroundings of Seoul (서울지역 생활환경주변의 공기 중 석면분포 특성에 관한 연구)

  • Lee, Jinhyo;Lee, Suhyun;Kim, Jihui;Oh, Seokryul;Shin, Jinho;Eom, Seokwon;Chae, Youngzoo;Lee, Jinsook;Koo, Jayong
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.9
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    • pp.613-623
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    • 2013
  • This study is purposed to evaluate the airborne asbestos concentrations in life environment surroundings in Seoul. In study, we investigated airborne asbestos concentrations in thirteen subway stations, four monitoring networks and each vicinity roadside, six stream surroundings, four tunnels quarterly and we also investigated relationship between the airborne asbestos concentrations and ambient temperature in monitoring networks and time-based airborne asbestos concentration variability for two typical monitoring networks, two subway stations transferred and used by lots of people through Phase Contrast Microscopy (PCM) and Transmission Electron Microscopy (TEM). The airborne asbestos concentrations by PCM for 4 objects of study were less than the detection limit (7 fiber/$mm^2$) in 111 (50%) out of 223 samples. The highest concentration was 0.0130 f/cc. But additional TEM analysis result for samples exceeding the guideline value for indoor air quality (0.01 f/cc) proposed by the Ministry of Environment (Korea), no asbestos was detected. Similarly TEM analysis result for 124 samples, no asbestos was detected. The average airborne asbestos concentrations by PCM in subway stations, monitoring networks, streams and tunnels were $0.0041{\pm}0.0027$ f/cc, $0.0015{\pm}0.0011$ f/cc, $0.0024{\pm}0.0012$ f/cc and $0.0016{\pm}0.0020$ f/cc. All objects of study were satisfied with the guideline value for indoor air quality. The relationship between the airborne asbestos concentrations and ambient temperature in monitoring networks was generally positive correlation (r = 0.660). The higher ambient temperature was and the more transient population was, the airborne asbestos concentrations by time for two subway stations were increased. While the airborne asbestos concentrations for two monitoring networks showed no variation pattern according to time.

Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.165-172
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    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

Asbestos Exposure and Health Risk Assessment for the Residents Near the Abandoned Mining Area in Chungbuk, Korea (충북지역 폐석면광산 인근주민의 석면노출과 건강위해도 평가)

  • Shin, Jin-Ho;Oh, Seok-Ryul;Hwang, Soon-Yong;Chung, Sook-Nye;Kim, Ji-Hui;Nam, Eun-Jung;Lee, Jin-Hyo;Choi, Hee-Jin;Eom, Seok-Won;Chae, Young-Zoo;Park, Chul-Hwi
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.5
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    • pp.345-350
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    • 2012
  • To investigate the exposure and health risk assessment for the residents near the D-asbestos mine in Chungbuk, Korea. We analyzed asbestos in the 20 ambient air and 23 activity based samples near the mine. The airborne sample results are showed that 8 of 20 samples ranged between 0.0025 to 0.0029 f/cc (fiber per cubic centimeter) and the others were below the detection limit by phase contrast microscopy (PCM). In addition, asbestos fibers were under the detection limit or not being by transmission electron microscopy (TEM). Based on interview and survey targeting the local residents, we made the activity based sampling (ABS) scenarios fit to the conditions of field. At the same time, we calculated the excess lifetime cancer risk (ELCR) of these ABS scenarios according to the ELCR average value and 95% upper confidence limit (UCL). At the case of weed whacking, soil digging and sweeping yard scenario, 95% UCL of ELCR exceeded the $1{\times}10^{-4}$, acceptable risk range for exposure. Based on our study results, it is necessary safety measures such as risk communication, abatement or management of naturally occurring asbestos (NOA).

Expression of Growth Factors and Secretory Leukocyte Protease Inhibitor (SLPI) in RAW264.7 Cells after Lipopolysaccharide (LPS) Stimulation (대식세포주 (RAW264.7)에서 박테리아성 지질다당류 (LPS) 자극에 의한 분비백혈구단백분해효소억제제 (SLPI)와 성장인자들의 발현)

  • Son, Wook-Hee;Choi, Baik-Dong;Jeong, Soon-Jeong;Wang, Guan-Lin;Hwang, Ho-Keel;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.2
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    • pp.93-102
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    • 2007
  • Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

Establishment of Mouse Embryonic Stem Cell-like Cells from In Vitro Fertilized Embryos (체외수정 생쥐 배아에서의 배아 줄기세포 확립)

  • Shin, Yong-Moon;Park, Yong-Bin;Kim, Hee-Sun;Oh, Sun-Kyung;Chun, Dae-Woo;Suh, Chang-Suk;Choe, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.1-12
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    • 2002
  • Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.