• Title/Summary/Keyword: Dye

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Purinergic regulation of calcium signaling and exocytosis in rat prostate neuroendocrine cells

  • Kim, Jun-Hee;Kim, Mean-Hwan;Koh, Duk-su;Park, So-Jung;Kim, Soo-Jung;Nam, Joo-Hyun;Lee, Jee-Eun;Uhm, Dae-Yong;Kim, Sung-Joon
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.54-54
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    • 2003
  • Prostate gland contains neuroendocrine cells (PNECs) are playing important roles in physiological and pathophysiological processes of the prostate gland. Here, we investigated the role of purinoceptors in PNECs freshly isolated from rat ventral prostate (RPNECs) that show immunoreactivity to chromogranin A. Fura-2 ratiometry revealed that ATP evokes both fast Ca$\^$2+/ influx and store Ca$\^$2+/ release in RPNECs. A whole-cell patch clamp study demonstrated fast inactivating cationic current activated by ATP or by ${\alpha}$,${\beta}$-MeATP, which was blocked by ATP-TNP. The activation of P2X inward current was tightly associated with a sharp increase in [Ca$\^$2+/]$\sub$c/. The presence of P2X1/3 subtypes were proved by RT-PCR analysis. For the stored Ca$\^$2+/ release, ATP and UTP showed similar effects, suggesting the dominant role or P2Y2 subtypes, also confirmed by RT-PCR. Both P2X (${\alpha}$,${\beta}$-MeATP) and P2Y (UTP) stimulation induced changes in the cell morphology (initial shrinkage and blob formation on the surface) reversibly. Exocytotic membrane trafficking events were monitored with the membrane-bound fluorescent dye, FM1-43 using confocal microscopy. In spite of the similar Ca$\^$2+/ responses, UTP was far less effective in triggering exocytosis than ${\alpha}$,${\beta}$ -MeATP. Since serotonin is reportedly stored in the secretory granule of PNECs, we directly examined whether the aforementioned agonists elicit release of serotonin using carbon fiber electrode-amperometry. In accordance with the results of FM1 -43 experiments, ${\alpha}$,${\beta}$-MeATP efficiently evoke serotonin secretion while not with UTP. In summary, the P2X-mediated Ca$\^$2+/ influx plays crucial roles in the exocytosis of RPNECs. Although a global increase in [Ca$\^$2+]$\sub$c/ might be related with the morphological changes, a sharp rise of [Ca$\^$2+/]$\sub$c/ in the putative sub-plasmalemmal ‘microdomains’ might be a decisive factor for the exocytosis.

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The Role of Glial Cells in Regenerative Responses of the Injured Corticospinal Tract Axons in Rats Treated with Cindii Rhizoma (흰쥐의 척수손상 후 천궁처리에 의한 피질척수로 축삭재생 반응시 Glial cells의 역할)

  • Han, Yeong-Su;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.3
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    • pp.19-39
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    • 2008
  • Objectives : It has been reported that CG was effective in decreasing injury to neural tissues. To investigate neural responses in the injured spinal cord, an extract of CG was examined to determine its effect on neural responses in the injured spinal cords of rats. Methods : After CG treatment was applied to the spinal cord of rats given a contusion injury, the re-growth responses of injured neural tissues and corticospinal tract axons was observed by measuring the number of GAP-43, Cdc2, and phospho-Erk1/2 proteins, CST axons, GFAP-stained astrocytes, and Glial scarring in the injured spinal cord. Results : Levels of GAP-43, Cdc2, and phospho-Erk1/2 proteins were found to have increased in the injured spinal cord region. The number of GFAP-stained astrocytes also increased within and around the injury cavity. Glial scarring, which was identified by CSPG immunofluorescence staining, was reduced by CG treatment. Anterograde tracing by Dil dye showed that the elongation of the CST axons in the dorso-medial white matter area was almost completely prevented at the injury site. Collateral sprouting was observed in the spinal cord rostrally close to the injury site, and CG treatment further increased axonal arborization in the corresponding region. In vivo migration of CST axons and astrocytes using an implanted polymer tube system showed more of an increase in enhanced migration of axons and astrocytes in CG-treated group compared to the injury control group. Conclusions : These results suggest that CG activated neural responses - including astrocyte migration - and promotes axonal regenerative activity in the injured spinal cord area.

EVALUATION OF THREE FLOWABLE COMPOSITES AS PIT AND FISSURE SEALANTS WITHOUT USING BONDING AGENTS (접착제 미 사용시 치면열구전색제로서의 유동성 복합레진 평가)

  • Kwon, Ho-Beom;Kim, Myoung-Jin;Shin, Cheol-Hwan;Kim, Ji-Yeon;Park, Ki-Tae
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.244-252
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    • 2006
  • The aims of this study were to determine if flowable composites can be used as pit and fissure sealants without bonding agents. Three flowable composites(Filtek Flow, Tetric Flow, Charmfil Flow) and a filled sealant (Ultraseal XT plus) were used. The pattern of resin tag formation in the four sealant materials were compared using SEM. For the microleakage assessment, 54 extracted human premolar teeth were randomly divided into 3 groups containing 18 premolars each. In each group, a conventional filled sealant and one of the three flowable composites were applied to occlusal fissures. The teeth were thermocycled(1200 cycles between $5^{\circ}{\pm}2^{\circ}C\;and\;55^{\circ}{\pm}2^{\circ}C$ with a dwell time of 30 seconds) and immersed in a 1% methylene blue solution for 48 hours. Each tooth was sectioned and examined to determine the extent of dye penetration. Three flowable composites and a filed sealant showed a similar resin tag formation pattern. The three flowable composites showed significantly more microleakage in each group than the filled sealant. The level of microleakage was similar in the three flowable composites. Flowable composites are not recommended as pit and fissure sealants because more microleakage can occur even when occlural fissures are mechanically widened.

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Fabrication of Chemical Sensors for the Detection of Acidic Gas using 1,3-bisdicyanovinylindane (1,3-bisdicyanovinylindane을 이용한 산성가스 감지용 화학 센서 제작)

  • Song, Hwan-Moon;Park, Young-Min;Son, Young-A;Lee, Chang-Soo
    • Korean Chemical Engineering Research
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    • v.46 no.1
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    • pp.184-188
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    • 2008
  • This study presented simple and efficient fabrication of chemical sensors for the detection of acidic gas using 1,3-bisdicyanovinylindane as an indicator because it can be promising materials having property of the rapid color change according to the variation of pH. The dissociation of proton and dye in acidic condition as changing of ion pairs give rise to dramatically change the absorbance intensity of 1,3-bisdicyanovinylindane, which can be easily applied to the development of chemical sensors. In addition, indicator dyes having negatively charge in aqueous phase can be easily fabricated using layer-by-layer (LBL) methods by way of electrostatic interaction. For the proof of concept, we demonstrated the abrupt presentation of skeleton symbol on the chemical sensor, which could be resulted from the reaction of 1,3-bisdicyanovinylindane as background color with acidic gas. Thus, the rapid appearance of symbol will induce user's caution under the emergency condition. The presented chemical gas sensor using 1,3-bisdicyanovinylindane have strong advantages. First, the fabrication process of gas sensor was very simple and low-cost. Secondly, sensors reacted by acidic gas could be reused for several times. Finally, the chemical gas sensor would be environmentally friend, which can be a basic tool for the realization of eco-organic sensor device.

The Flora of Gwangchiryeong Area Adjacent to the DMZ (DMZ에 인접한 광치령 일대의 식물상)

  • Heo, Tae-Im;Shin, Hyun-Tak;Kim, Sang-Jun;Lee, Jun-Woo;Jung, Su-Young;An, Jong-Bin
    • Korean Journal of Environment and Ecology
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    • v.31 no.1
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    • pp.1-23
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    • 2017
  • Gwangchiryeong, located adjacent to the Korean DMZ, is a mountain pass connecting Yang-Gu, Gangwon-do with In-je. It reaches a height of 800m (2,624ft) and stands on the flank of Mt. Daeam-san, which has an elevation of 1,304m (4,278ft). It is important to survey the flora of Gwangchiryeong in that not only the area encompassing the waterfall and the valley along with dense old-growth forest has various plant species but also it is a way to make out what kind of plant species exist in the DMZ due to the geographical characteristics of Gwangchiryeong adjacent to the Korean DMZ. Therefore, this study was conducted to elucidate the distribution of vascular plants and their value in Gwangchiryeong. The vascular plants that were collected 9 times from April 2014 to September 2015, and consisted of a total of 513 taxa: 88 families, 297 genera, 441 species, 4 subspecies, 62 varieties and 6 forms. Among those observed species, the rare plant species designated by Korea Forest Service were 22 taxa including Hanabusaya asiatica, Paeonia japonica, Saxifraga octopetala, etc. Endemic species were 22 taxa containing Cirsium setidens, Heloniopsis koreana, Salvia chanryoenica, etc. 18 taxa were recorded as naturalized plants and their naturalized rate and urban index were noted as 3.5% and 5.6%. Among the surveyed 513 taxa, edible, medicinal, pasturing, ornamental, timber, dye, fiber and industrial plants were 197 taxa, 154 taxa, 180 taxa, 49 taxa, 24 taxa, 8 taxa, 4 taxa and 3 taxa, respectively.

Effects of Ginseng Saponins on the Induction of Differentiation in Mammary Epithelial Cells and Mammary tumor Cells (홍삼 사포닌에 의한 유선상피 및 유선암세포의 분화 유도 효과 연구)

  • 오미숙;백기주;전성실;김규원;최강주;김남득
    • Journal of Ginseng Research
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    • v.24 no.4
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    • pp.188-195
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    • 2000
  • Using Ginseng saponins (crude saponin and total saponin) and ginsenoside Rbl Rb2, Rc, Rd, Re, Rhl, and Rh2 in this study, we have examined the effects of the compounds on the induction of differentiation in normal rat mammary epithelial cells and 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumor cells in culture. When normal rat mammary organoids were cultured in 100-mm culture plates in the presence or absence of ginseng saponins, there were four different cell colonies after two weeks in culture: cobble stone, spindle, honey comb, and senescence type colonies. Ginseng saponins showed different effects on the development of each colonies. Scrape-loading dye transfer tech-nique was performed to measure the effects of total saponin, Rhl, and Rh2 on intercellular junctional communication. Intercellular communication was not observed at short cultilral time, e.g., four or seven days, but when it cultured it up to two weeks, cell to cell communication was observed in saponin-treated cells. Reconstituted basement membrane, Matrigel, supported the growth and development several different multicellular structures from normal mammary organoids (e.g., ductal, webbed, stellate, and squamous colonies) or DMBA-induced mammary tumor (e.g., alveolar unit, foamy alveolar unit, squamous metaplasia, lobule-ductal, stellate, and webbed colony). In ginseng saponin-treated groups, webbed colonies were more and squamous colonies were less than control group. Moreover, the ductal colonies, marker tructure of well-differentiate mammary epithelial cells, were developed more in saponin-treated group than in control group. In conclusion, ginseng saponins affected on the differentiation of normal rat mammary epithelial cells and DMBA-induced mammary tumor cells in culture.

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FLUORESCENT LABELLING OF MC3T3 CELL LINE BY 5-(AND-6)-CARBOXY-2', 7'-DICHLOROFLUORESCEIN DIACETATE, SUCCINIMIDYL ESTER MIXED (MC3T3 preosteoblast cell line의 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed에 의한 fluorescent labelling)

  • Kook, Min-Suk
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.6
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    • pp.461-467
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    • 2005
  • Background. 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed (CFSE) is the fluorescent labelling agent of living cells and used to trace the cells in vivo after transplatnation of various cells. The CFSE labelled cells can maintain fluorescence for up to 7 days after labelling. The MC3T3-E1 cell line (MC3T3) has been used for many studies about osteoblast, which is well known as a mouse preosteoblast. So the CFSE would be used to trace the transplanted MC3T3. However there are few reports about CFSE labelling of MC3T3. This study is aimed to know about adequate concenturation and incubation time of CFSE to MC3T3. Materials and methods. The MC3T3 was incubated in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ using ${\alpha}$-minimal essential medium (${alpha}$-MEM) containing10% FBS and gentamycin. Ten mM CFSE solution in dimethylsulphoxide (DMSO: 1%) was diluted with phosphate buffered saline (PBS) and final concentration of culture medium was, respectively, 5, 10, 15, 20, 25 and 30 ${{\mu}M$. Then the MC3T3 was incubated with CFSE in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 and 45 minutes in each concentration. The fluorescence of CFSE labelled cells was analysed with a inverted fluorescence microscope. The duration of cell labelling was also studied. Trypan blue dye exclusion test was done for cell viability. Results. For concentration between 5 and 10 ${\mu}M$, CFSE did not significantly label the MC3T3 in vitro. The destruction of MC3T3 was observed at the concentration of 20 ${\mu}M$. In the concentration of 15 ${\mu}M$, the best labelling was obtained at an incubation period between 15 and 30 minutes. The MC3T3 labelled with an incubation period of 15 minutes at 15 ${\mu}M$ was still fluorescent 7 days after CFSE labelling. The mean cell viability was 95.93%. Conclusion. These results suggests an incubation period of 15 minutes at 15 ${\mu}M$ of CFSE provides best labelling of MC3T3 in vitro.

Preparation and characterization of Ga-doped TiO2 nanofibers by electrospinning (전기방사를 이용한 Ga이 첨가된 나노섬유의 제작 및 특성평가)

  • Song, Chan-Geun;Kang, Won Ho;Yoon, Jong-Won
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.6
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    • pp.274-278
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    • 2012
  • $TiO_2$ can be used optically and is applied on many areas such as gas sensor, solar cell and photocatalysis. Electrospun nanofibers have received great interest for development and utilization in some novel applications, such as chemical sensors, dye-sensitized solar cell and photo catalysis. In this study, pure $TiO_2$ and Ga-doped $TiO_2$ nanofibers synthesized by a modified electrospinning method. The Ga doped $TiO_2$ solution is prepared by mixing poly vinyl pyrrolidone, ethyl alcohol, and titanium (IV) isopropoxide. By electrospinning these sols, nanofibers were fabricated. These fibers are heat-treated at $800^{\circ}C$ in air. The prepared pure $TiO_2$ and Ga-doped $TiO_2$ nanofibers samples were characterized by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Raman spectroscopy.

Folk Plants in the Inland of Northern Area in Gangwon-do (강원도 내륙 북부지역의 민속식물)

  • Kim, Kyung-Ah;Han, Jun-Soo;Cheon, Kyeong-Sik;Jang, Jin-Hwan;Ok, Gil-Hwan;Yoo, Ki-Oug
    • Korean Journal of Plant Resources
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    • v.25 no.1
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    • pp.48-62
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    • 2012
  • This study was carried out to classify the folk plants in inland of northern region of Gangwon-do. According to the survey results from 35 places of 9 counties and cities from April to October, 2010, the folk plants in inland of northern region of Gangwon-do consisted of a total 261 taxa; 73 families, 185 genera, 225 species, 2 subspecies, 30 varieties, and 4 form. Among the investigated 261 taxa, 3 Korean endemic, 11 rare plants and 9 naturalized plants were also included. The use by usage of 261 taxa was: 378 times; Edible, 53 times; Medicinal, 8 times; Dye, 7 times; Ornamental, 6 times; Fuel, 5 times; Spice, 3 times; Aroma, 2 times; Nectar, 41 times; Others, respectively, so the edible use is the highest. The most useful part was the leaf, followed by whole plant and root. The consistency comparison between the scientific name and the local name were the highest in the 20's and the lowest in 80's.

The Regional Folk Plants in Southern Inland Area of Gyeonggi-do (경기남부 내륙지역의 민속식물자원)

  • Jeong, Hye-Ran;Choi, Kyung;Park, Kwang-Woo
    • Korean Journal of Plant Resources
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    • v.25 no.5
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    • pp.523-542
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    • 2012
  • This study was carried out to clarify the regional folk plants in the southern inland area of Gyeonggi-do Province. According to the survey results, derived from 1,229 sheets of 90 residents at 35 places from 9 counties and cities between October 2009 and December 2010, the folk plants in the inland southern area of Gyeonggi-do consisted of a total of 259 taxa; 82 families, 200 genera, 221 species, 4 subspecies, 32 varieties, and 2 forma. The classification of the plants by its usage was; 214 as edible, 119 as medicinal, 34 as ornamental, 18 as timber, 12 as dye, 5 as oil, 3 as spice, 2 as fiber, 2 as aroma, and 22 as others uses. So, the edible and medicinal uses are higher than other uses. The most useful part was the leaf, followed by fruit, stem, root, and flower. The collection season was focused on spring and summer, because the leaf was mainly collected in an edible condition during those seasons. Traditional knowledge of folk plants was mostly passed down orally. The consistency comparison between the standard common name and the local name was the highest in the people in 50s and the lowest in those in 80s.