• Title/Summary/Keyword: Fluorescent microscopy

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Green Synthesis of Multifunctional Carbon Nanodots and Their Applications as a Smart Nanothermometer and Cr(VI) Ions Sensor

  • Li, Lu;Shao, Congying;Wu, Qian;Wang, Yunjian;Liu, Mingzhu
    • Nano
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    • v.13 no.12
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    • pp.1850147.1-1850147.14
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    • 2018
  • In this work, water-soluble and blue-emitting carbon nanodots (CDs) were synthesized from apple peels for the first time via one-step hydrothermal method. The synthetic route is facile, green, economical and viable. The as-prepared CDs were characterized thoroughly by transmission electron microscopy (TEM), X-ray diffraction (XRD), Raman, Fourier transform infrared (FT-IR), X-ray photoelectron (XPS), fluorescence and UV-Vis absorption spectroscopy in terms of their morphology, surface functional groups and optical properties. The results show that these CDs possessed ultrasmall size, good dispersivity, and high tolerance to pH, ionic strength and continuous UV irradiation. Significantly, the CDs had fast and reversible response towards temperature, and the accurate linear relationship between fluorescence intensity and temperature was used to design a novel nanothermometer in a broad temperature range from 5 to $65^{\circ}C$ facilely. In addition, the fluorescence intensity of CDs was observed to be quenched immediately by Cr(VI) ions based on the inner filter effect. A low-cost Cr(VI) ions sensor was proposed employing CDs as fluorescent probe, and it displayed a wide linear range from 0.5 to $200{\mu}M$ with a detection limit of $0.73{\mu}M$. The practicability of the developed Cr(VI) sensor for real water sample assay was also validated with satisfactory recoveries.

Overexpression of ginseng cytochrome P450 CYP736A12 alters plant growth and confers phenylurea herbicide tolerance in Arabidopsis

  • Khanom, Sanjida;Jang, Jinhoon;Lee, Ok Ran
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.645-653
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    • 2019
  • Background: Cytochrome P450 enzymes catalyze a wide range of reactions in plant metabolism. Besides their physiological functions on primary and secondary metabolites, P450s are also involved in herbicide detoxification via hydroxylation or dealkylation. Ginseng as a perennial plant offers more sustainable solutions to herbicide resistance. Methods: Tissue-specific gene expression and differentially modulated transcripts were monitored by quantitative real-time polymerase chain reaction. As a tool to evaluate the function of PgCYP736A12, the 35S promoter was used to overexpress the gene in Arabidopsis. Protein localization was visualized using confocal microscopy by tagging the fluorescent protein. Tolerance to herbicides was analyzed by growing seeds and seedlings on Murashige and Skoog medium containing chlorotoluron. Results: The expression of PgCYP736A12 was three-fold more in leaves compared with other tissues from two-year-old ginseng plants. Transcript levels were similarly upregulated by treatment with abscisic acid, hydrogen peroxide, and NaCl, the highest being with salicylic acid. Jasmonic acid treatment did not alter the mRNA levels of PgCYP736A12. Transgenic lines displayed slightly reduced plant height and were able to tolerate the herbicide chlorotoluron. Reduced stem elongation might be correlated with increased expression of genes involved in bioconversion of gibberellin to inactive forms. PgCYP736A12 protein localized to the cytoplasm and nucleus. Conclusion: PgCYP736A12 does not respond to the well-known secondary metabolite elicitor jasmonic acid, which suggests that it may not function in ginsenoside biosynthesis. Heterologous overexpression of PgCYP736A12 reveals that this gene is actually involved in herbicide metabolism.

Viator vitreocola gen. et sp. nov. (Stylonematophyceae), a new red alga on drift glass debris in Oregon and Washington, USA

  • Hansen, Gayle I.;West, John A.;Yoon, Hwan Su;Goodman, Christopher D.;Goer, Susan Loiseaux-de;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.34 no.2
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    • pp.71-90
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    • 2019
  • A new encrusting red alga was found growing abundantly on glass debris items that drifted ashore along the coasts of Oregon and Washington. These included discarded fluorescent tubes, incandescent light bulbs, capped liquor bottles, and ball-shaped fishing-net floats. Field collections and unialgal cultures of the alga revealed that it consisted of two morphological phases: a young loosely aggregated turf and a mature consolidated mucilaginous crust. The turf phase consisted of a basal layer of globose cells that produced erect, rarely branched, uniseriate to multiseriate filaments up to $500{\mu}m$ long with closely spaced cells lacking pit-plugs. These filaments expanded in size from their bases to their tips and released single cells as spores. At maturity, a second phase of growth occurred that produced a consolidated crust, up to $370{\mu}m$ thick. It consisted of a basal layer of small, tightly appressed ellipsoidal-to-elongate cells that generated a mucilaginous perithallial matrix containing a second type of filament with irregularly spaced cells often undergoing binary division. At the matrix surface, the original filaments continued to grow and release spores but often also eroded. Individual cells, examined using confocal microscopy and SYBR Green staining, were found to contain a central nucleus, a single highly lobed peripheral chloroplast without a pyrenoid, and numerous chloroplast nucleoids. Morphological data from field and culture isolates and molecular data (rbcL, psbA, and SSU) show that this alga is a new genus and species which we name Viator vitreocola, "a traveller on glass."

Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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Effects of Horse Bone Powder Extract on Longitudinal Bone Growth in Adolescent Male Rats (마골(馬骨) 추출물의 성장기 흰쥐 장골 길이 성장에 대한 효과)

  • Lee, Se-Na;Choi, Han;Son, Jae-Bong;Jeong, Jae-Seung;Kim, Yun-Jung;Kang, Dae-Pyung;Park, Soo-Yeong;Kang, Se-Chan;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.24 no.3
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    • pp.69-77
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    • 2009
  • Objectives : This study was designed to investigate the effects of horse bone powder extract on the growth of longitudinal bone in adolescent male rats. Methods: Longitudinal bone growth was measured by fluorescent microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate, the induction of local insulin-like growth factor-1 (IGF-1), and bone morphogenetic protein-2 (BMP-2) were measured. Results: Horse bone powder extract enhanced longitudinal bone growth and total height of the growth plate. Also, it promoted the induction of local IGF-1 and BMP-2 of the growth plate. Conclusions: This study shows that the horse bone powder extract effects longitudinal bone growth in adolescent rats and might be used for both stunted adolescents and inherent growth failure patients.

Effects of Cheunggyeongsamul-tang Extract on Longitudinal Bone Growth in Adolescent Female Rats (청경사물탕(淸經四物湯) 추출물의 성장기 흰쥐 장골길이 성장에 대한 효과)

  • Lee, Jung-Hun;Lee, Se-Na;Son, Jae-Bong;Keum, So-Hyun;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.149-157
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    • 2009
  • Objectives : This study was designed to investigate the effects of Cheunggyeongsamul-tang extract on the growth of longitudinal bone in adolescent female rats. Methods : Longitudinal bone growth was measured by fluorescent microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate, the induction of local insulin-like growth factor-1 (IGF-1), IGF-1 receptor, bone morphogenetic protein-2 (BMP-2), BMPR-1A, indian hedgehog (IHH), and parathyroid hormone-related protein (PTH-rP) were measured. Results : Cheunggyeongsamul-tang extract enhanced longitudinal bone growth and total height of the growth plate. Also, it promoted the induction of local IGF-1, BMP-2, IHH and PTH-rP of the growth plate. Conclusions : This study shows that the Cheunggyeongsamul-tang extract effects longitudinal bone growth in adolescent rats and might be used for both stunted adolescents and inherent growth failure patients.

Particle-motion-tracking Algorithm for the Evaluation of the Multi-physical Properties of Single Nanoparticles (단일 나노입자의 다중 물리량의 평가를 위한 입자 모션 트랙킹 알고리즘)

  • Park, Yeeun;Kang, Geeyoon;Park, Minsu;Noh, Hyowoong;Park, Hongsik
    • Journal of Sensor Science and Technology
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    • v.31 no.3
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    • pp.175-179
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    • 2022
  • The physical properties of biomaterials are important for their isolation and separation from body fluids. In particular, the precise evaluation of the multi-physical properties of single biomolecules is essential in that the correlation between physical and biological properties of specific biomolecule. However, the majority of scientific equipment, can only determine specific-physical properties of single nanoparticles, making the evaluation of the multi-physical properties difficult. The improvement of analytical techniques for the evaluation of multi-physical properties is therefore required in various research fields. In this study, we developed a motion-tracking algorithm to evaluate the multi-physical properties of single-nanoparticles by analyzing their behavior. We observed the Brownian motion and electric-field-induced drift of fluorescent nanoparticles injected in a microfluidic chip with two electrodes using confocal microscopy. The proposed algorithm is able to determine the size of the nanoparticles by i) removing the background noise from images, ii) tracking the motion of nanoparticles using the circular-Hough transform, iii) extracting the mean squared displacement (MSD) of the tracked nanoparticles, and iv) applying the MSD to the Stokes-Einstein equation. We compared the evaluated size of the nanoparticles with the size measured by SEM. We also determined the zeta-potential and surface-charge density of the nanoparticles using the extracted electrophoretic velocity and the Helmholtz-Smoluchowski equation. The proposed motion-tracking algorithm could be employed in various fields related to biomaterial analysis, such as exosome analysis.

Characterization of Nitric Oxide (NO)-Induced Cell Death in Lung Epithelial Cells (폐상피세포에서 Nitric Oxide (NO)에 의한 세포사에 관한 연구)

  • Yong, Wha Shim;Kim, Youn Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.2
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    • pp.187-197
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    • 2004
  • Background : Nitric Oxide (NO) is a multi-faceted molecule with dichotomous regulatory roles in many areas of biology. NO can promote apoptosis in some cells, whereas it inhibits apoptosis in other cell types. This study was performed to characterize NO-induced cell death in lung epithelial cells and to investigate the roles of cell death regulators including iron, bcl-2 and p53. Methods : A549 cells were used for lung epithelial cells. SNP (sodium nitroprusside) and SNAP (S-nitroso-N-acetyl- penicillamine) were used for NO donor. Cytoxicity assay was done by MTT assay and crystal violet assay. Apoptotic assay was done by fluorescent microscopy after double staining with propidium iodide and hoecst 33342. Iron inhibition study was done with RBCs and FeSO4. For bcl-2 study, bcl-2 overexpressing cells (A549-bcl-2) were used and for p53 study, Western blot analysis and p53 functionally knock-out cells (A549-E6) were used. Results : SNP and SNAP induced dose-dependent cell death in A549 cells and fluorescent microscopy revealed that SNAP induced apoptosis in low doses but necrosis in high doses while SNP induced exclusively necrotic cell death. Iron inhibition study using RBCs and FeSO4 significantly blocked SNAP-induced cell death. And also SNAP-induced cell death was blocked by bcl-2 overexpression. Finally, we found that SNAP activate p53 by Western blot analysis and that SNAP-induced cell death was decreased in the abscence of p53. Conclusion : In lung epithelial cells, NO can induce cell death, more precisely apoptosis in low doses and necrosis in high doses. And iron, bcl-2, and p53 play important roles in NO-induced cell death.

Study on the Copper-Arsenic Green Pigments used on Shamanic Paintings in the 19~20th century (19~20세기 무신도 등에 사용된 구리-비소 녹색 안료에 대한 연구)

  • Oh, Joon Suk;Choi, Jung Eun;Choi, Yoon Hee
    • Journal of Conservation Science
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    • v.31 no.3
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    • pp.193-214
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    • 2015
  • The green pigments of shamanic paintings (83 items) in the 19~20th century were analyzed with X-ray fluorescent spectrometer(XRF), scanning electron microscopy (SEM) and fourier transform infrared spectroscopy (ATR-FTIR). In 60 items, copper and arsenic were detected in green pigments of the paintings by XRF spectra. Cu $K{\alpha}$/As $K{\beta}$ (peak intensity ratio of copper and arsenic) of shamanic paintings of Bokgaedang (shrine), solemn paintings (romance of three kingdoms) of Donggwanwangmyo and unknown enshrined place were 5.93~12.04 and higher compared to 5.67-6.26 of standard emerald greens and 4.01~7.89 of remaining shamanic paintings. The SEM images of crystal forms of copper-arsenic green pigments were various. Crystal forms were divided into oval and round spherulite with intersecting plate crystals and spherulite with agglomerate plate crystals. The crystals of the latter were found in shamanic paintings of Bokgaedang (shrine), solemn paintings (romance of three kingdoms) of Donggwanwangmyo and unknown enshrined place and the former were found in the rest of shamanic paintings. Copper-arsenic green pigments of shamanic paintings were identified as Scheele's green from shamanic paintings of Bokgaedang (shrine) and romance of three kingdoms. Emerald green from the rest of shamanic paintings by ATR-FTIR. From analytical results, it is confirmed that Scheele's green of shamanic paintings of Bokgaedang and romance of three kingdoms was used in the 1850s~1870s and emerald green had been widely used from late 19th century to 1970 in the rest of shamanic paintings.

Development of aortic endothelial cells to express CD37 and CD73 isolated from alpha 1,3-galactosyltransferase knock-out and MCP expressing pig (alpha 1,3-galactosyltransferase 기능 제거 및 MCP 발현 형질전환 돼지의 대동맥 혈관내피세포에 CD37/CD73 발현 세포주 개발)

  • No, Jin-Gu;Byun, Sung-June;Yang, Hyeon;Ock, Sun A;Woo, Jae-Seok;Lee, Hwi-Cheul;Hwang, In-sul;Kim, Ji-Youn;Park, Sang Hyoun;Lee, Joo Young;Oh, Keon Bong
    • Journal of Embryo Transfer
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    • v.33 no.3
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    • pp.129-137
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    • 2018
  • Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig ($GalT^{-MCP/-MCP}$). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain $GalT^{-MCP/-MCP}/CD39/CD73$ pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from $GalT^{-MCP/-MCP}$ pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate $GalT^{-MCP/-MCP}/CD39/CD73$ pig expressing CD39 and CD73 at endothelial cells.