• Title/Summary/Keyword: Fluorochrome

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Rapid Assesment of Microspore Development Stage in Pepper Using DAPI and Ferric chloride

  • Kim, Moon-Za;Jang, In-Chang
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.129-134
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    • 2000
  • Clear visualization of pepper (Capsicum annuum L.) microspore nuclei with common stains such as acetocarmine or propionocarmine is difficult, hindering cytological analysis. The DAPI stain after the addition of ferric chloride solution to fixative resulted in clear visualization of nuclei. For clear visualization of nuclei and slight fluorescence of microspore wall, addition of 40-60 ${mu}ell$ of ferric chloride solution to the 1 $m\ell$ fixative was identified as most effective. At all stages of gametophytic development, the nuclei can be distinctly visualized. Starch granules does not intefere with the fluorochrome, and so the vegetative and generative nuclei were cleary visible in binucleate pollens. With its rapidity and reliability, this technique represents an efficient tool for routine staging or investigation of the nuclear status of the microspore during culture.

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High Resolution G- and R-banding for Accurate analysis in Human Chromosomes (사람의 정확한 염색체 분석을 위한 고해상 G-분염 염색체와 R-분염 염색체)

  • Park, So-Yun;Choi, Soo-Kyung;Lee, Hyung-Hoan
    • Clinical and Experimental Reproductive Medicine
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    • v.22 no.2
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    • pp.109-122
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    • 1995
  • A technique is described for producing high resolution G- and R-banded chromosomes in human peripheral lymphocyte cultures. Cultured lymphocyte cells were exposed to ethidium bromide ($10{\mu}g/ml$) and colcemid ($0.02{\mu}g/ml$) each for 2.5h and 0.5h prior to harvest for high resolution G-banded chromosomes. High resolution R-band patterns were obtained by BrdU substitution which was revealed by the fluorochrome-photolysis-Giemsa staining technique. These methods are easy to perform and highly reliable. The data on relative length of chromosomes at the four mitotic stages are presented in units of percentage of haploid autosome length. The characteristic patterns of GTG-bands (G-bands after trypsin and Giemsa) and RBG bands (R-bands after BrdU and Giemsa) were analyzed.

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Heterosigma akasiwo의 핵형분석을 통한 생활사 연구를 위한 DAPI이용 기법

  • Lee Ju Yeon;Han Myeong Su
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2003.11a
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    • pp.121-124
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    • 2003
  • The goals of this study is to elucidate life cycle and to detect genetic differences within a single species of Heterosigma akashiwo. To elucidate life cycle of H. akashiwo, have to study of benthic stage and vegetative cell. So we studied identification of H. akashiwo cyst. The relative contents of DNA in nuclei were determined in Heterosigma akashiwo. Different stages of the life history were obtained from culture and natural sediments, and examined by microfluorometry after staining with the DNA-specific fluorochrome 4'-6-dianudubi-2-phenylindole(DAPI). Large cells mainly in exponensial stage, while small cell, pre-encystment cells(\ulcorner\ulcorner), showed in the end of the late growth stage. Type of DNA content showed the different with growth stage. Usually the small cell has the high level of IOD.

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THE HISTOLOGIC STUDY OF BONE HEALING AFTER HORIZONTAL RIDGE AUGMENTATION USING AUTO BLOCK BONE GRAFT (자가골 블럭 이식을 이용한 수평골 증강술시 이식골의 치유)

  • Oh, Jae-Kwen;Choi, Byung-Jun;Lee, Baek-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.3
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    • pp.207-215
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    • 2009
  • Purpose: The aim of the present study is to evaluate the long term bone healing after horizontal ridge augmentation using auto block bone graft for implant installation timing. Materials and Methods: Five Beagle dogs(which were 14 months old and weighted approximately 10kg). In surgery 1(extraction & bone defect), premolars(P2, P3,P4) were extracted and the buccal bone plate was removed to create a horizontally defected ridge. After three months healing, in surgery 2(ridge augmentation). Auto block bone grafts from the mandibular ramus were used in filling the bone defects were fixed with stabilizing screws. The following fluorochrome labels were given intravenously to the beagle dogs: oxytetracycline 1week after the surgery, alizarin red 4 weeks after the surgery, calcein blue 8 weeks after the surgery. The tissue samples were obtained from the sacrificed dogs of 1, 4, 8, 12, 16 weeks after the surgery. Non-decalcified sections were prepared by resin embedding and microsection to find thickness of $10{\mu}m$ for the histologic examination and analysis. Results: 1. We could achieve the successful reconstruction of the horizontal bone defect by auto block bone graft. The grafted bone block remained stable morohologically after 16 weeks of the surgery. 2. In the histologic view. We observed osteoid tissue from the sample $4^{th}$ week sample and active capillary reconstruction in the grafted bone from the $12^{th}$ week sample. Healing procedures of auto bone grafts were compared to that of the host bone. 3. Bone mineralization could be detected from the $8^{th}$ week sample. 4. Fluorochrome labeling showed active bony changes and formation at the interface of the host bone and the block graft mainly. Bony activation in the grafted bone could be seen from the $4^{th}$ week samples. Conclusions: Active bone formation and remodeling between the grafted bone and host bone can be seen through the revascularization. After the perfect adhesion to host bone, Timing of successful implant installation can be detected through the ideal ridge formation by horizontal ridge augmentation.

THE EFFECT OF AUTOLOGOUS PLATELET-RICH PLASMA (PRP) ON BONE FORMATION AROUND DENTAL IMPLANT IN THE RABBIT: A HISTOMOR-PHOMETRIC AND REMOVAL TORQUE STUDY

  • Yi Yang-Jin;Yang Jae-Ho;Lee Sun-Hyung;Kim Yung-Soo;Kwon Sang-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.39 no.6
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    • pp.659-681
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    • 2001
  • Platelet-rich plasma(PRP) has been known to increase the rate and degree of bone formation by virtue of growth factors in concentrated platelets. Although its great healing effect on bone defect or pre-implantation site preparation in conjunction with bone substitute has been reported, the effect associated with implant is unknown. The purpose of this study was to investigate the effect of PRP on rapid osseointegration of endosseous dental implants in the rabbit tibiae. Twenty two adult female New Zealand white rabbits, weighing approximately 2.7-3.3kg, were used for this study. Twelve of the 22 animals were used for histomorphometric analysis and ten of the 22 were for removal torque test. Each animal received two implants in each tibia (two treated with PRP and two as control) and was given fluorochrome intramuscularly. For histomorphometric analysis, rabbits were divided into four groups according to the healing period. At 1 week, 2 weeks, 4 weeks and 8 weeks postoperatively, each three animals were sacrificed serially and the amount and rate of bone formation around dental implant were examined on the undecalcified sections under fluorescent microscope, polarized microscope and light microscope connected to a personal computer equipped with image analysis system. For removal torque test, rabbits were divided into two groups and removal torque tests were performed at 4 weeks, 10 weeks after implant placement. In total, 88 screw shaped, commercially pure titanium implants (Neoplant, Neobiotech, Seoul, Korea) were used in this study. Labeling pattern reflected differences of two groups in bone formation rate at each period. Histomorphometrically, PRP group showed significantly higher bone volume within threads compared to control group at 2 weeks ($70.30{\pm}4.96%$ vs. $50.68{\pm}6.33%$; P < .01) and 4 weeks ($82.59{\pm}5.94%$ vs. $72.94{\pm}4.57%$; P < .05 ). PRP group at 1, 2 and 4 weeks revealed similar degree of bone volume formation comparable to control group at 2, 4 and 8 weeks, respectively. On the other hand, while PRP group showed higher bone-implant contact ($47.37{\pm}8.09%$) than control group ($33.16{\pm}13.47%$) at 2 weeks, there were no significant differences between PRP group and control group for any experimental period. Removal torque values also showed no significant differences between PRP group and control group at any experimental period (P > .05). These findings imply that PRP could induce rapid, more bone formation around implant during early healing period and get faster secondary stability for reducing healing period, though it has not induced bone maturation enough to resist functional loading.

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In Vitro Development of Reconstructed Bovine Embryos and Fate of Donor Mitochondria Following Nuclear Injection of Cumulus Cells

  • Do, Jeong-Tae;Lee, Bo-Yon;Kim, Seung-Bo;Lee, Hoon-Taek;Chung, Kil-Saeng
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.11-11
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    • 2001
  • In this study, we examined the developmental potential of reconstructed bovine embryos and the fate of donor mitochondria during their preimplantation development after nuclear transfer. Isolated cumulus cells were used as donor cells in nuclear transfer. Cumulus cells labelled with MitoTracker Green FM fluorochrome were injected into enucleated bovine MII oocytes and cultured in vitro. MitoTracker labelling on donor cells did not have a detrimental effect on blastocyst formation following nuclear transfer. Cleavage rate was about 69%(56/81) and blastocyst formation rate was 6.2% (5/81) at 7 days after nuclear transfer. The labelled mitochondria dispersed to the cytoplasm and became distributed among blastomeres and could be identified up to the 8- to 15-cell stages. Small patches of mitochondria were detected in some 8- to 15-cell stage embryos (5/20). However, donor mitochondria were not detected in embryos at the 16-cell stage and subsequent developmental stages. In the control group, mitochondria could be identified in arrested 1-cell embryos up to 7 days after nuclear transfer These results suggest that donor mitochondria disappear from recipient cytoplasm before 16-cell stage following nuclear transfer in reconstructed bovine embryos.

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A New Rhodamine B-coumarin Fluorochrome for Colorimetric Recognition of Cu2+ and Fluorescent Recognition of Fe3+ in Aqueous Media

  • Tang, Lijun;Li, Fangfang;Liu, Minghui;Nandhakumar, Raju
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3400-3404
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    • 2011
  • A new rhodamine B-coumarin conjugate (1) capable of recognizing both $Cu^{2+}$ and $Fe^{3+}$ using two different detection modes have been designed and synthesized. The metal ion induced optical changes of 1 were investigated in $CH_3CN-H_2O$ (1:1, v/v, HEPES 50 mM, pH = 7.0) solution. Sensor 1 exhibits selective colorimetric recognition of $Cu^{2+}$ and fluorescent recognition of $Fe^{3+}$ with UV-vis and fluorescence spectroscopy, respectively. Moreover, both of the $Cu^{2+}$ and $Fe^{3+}$ recognition processes are observed to be barely interfered by other coexisting metal ions.

Distribution of Mycoplasma in Witches'-broom Infected Jujube Tissue (빗자루병(病)에 감염(感染)된 대추나무 조직내(組織內)의 마이코플라스마 분포(分布))

  • La, Yong Joon;Lee, Deok Jae
    • Journal of Korean Society of Forest Science
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    • v.67 no.1
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    • pp.28-30
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    • 1984
  • Distribution of mycoplasma-like organisms (MLO) in the phloem tissue of witches'-broom infected jujube trees was investigated by fluorescence microscopy applying new fluorochrome DAPI (4'-6-diamidino-2-phenylindole, 2HCl). MLO were detected from the phloem of leaf and stem sections of diseased branches exhibiting typical witches'-broom symptom but not detected from those of symptomless branches of an infected tree. MLO were also present in the healthy looking leaves frequently found in the diseased shoots. Fluorescence microscopy revealed the presence of MLO in the phloem of root sections of infected trees.

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Effects of Daturae Flos and Daturae Semen Extract on the Immunocyte Response in Mice (만타라화(曼陀羅花) 및 만타라자(曼陀羅子) 수추출물(水抽出物)이 마우스의 면역세포기능(免疫細胞機能)에 미치는 영향(影響))

  • Ko, Woon-Che;Song, Ho-Joon;Shin, Min-Kyo
    • Korean Journal of Pharmacognosy
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    • v.21 no.4
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    • pp.307-316
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    • 1990
  • This study was undertaken to test the effects of Daturae Flos(DF) and Daturae Semen(DS) on the cellular and humoral immune responses, and the functions of the cells involved in immunoinflammation. Both extracts decreased the activity of superoxide dismutase, and the decrease was greater in the mouse group which was treated with DS. Both extracts decreased the phagocytic activity as measured by assessing the number of the latex particle within the phagocyte after incubation of peritoneal macrophages with fluorochrome-labelled latex particle and decreased natural killer cell activity as measured by enumerating the viable YAC-1 cells after treatment of target cells with splenic natural killer cells. Both extracts also decreased the cell-mediated immunity in vivo as assessed by measuring the ear thickness after sensitization and challenge with dinitrofluorobenzene, however, had no effects on the humoral immune responses as measured by checking hemolysin and hemagglutinin titers after immunization with sheep red blood cells(SRBC). Extracts of Semen caused decrease in the number of rosette forming cells between the splenic cells and SRBC. The results of this study suggested that both Daturae extracts could depress the immunoinflammation by affecting the various cell types involved in inflammation.

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A Study on the Separation of X- and Y- Spermatozoa in Farm Animals (가축에 있어서 X-정자와 Y-정자의 분류에 관한 연구)

  • 고대환;박흠대;정길생
    • Korean Journal of Animal Reproduction
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    • v.3 no.1
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    • pp.41-47
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    • 1979
  • This experiment was carried out to clarify the methods of the F-body test in human and the B-body test in buil and hog. The effect of pH and albumin concentration on the migration of X- and Y- sperm was also investigated. The results obtained were summarized as follows: 1. In the human semen, the frequency of sperm in which an F-body is visible was different by the fluorochrome. Namely, in case of quinacrine mustard, the F-body frequency was 48.8∼43.4 percent (average 49.6%), and in case of quinacrine dihydrochloride, that was 40.7∼50.8 percent (average 42.0%). 2. The frequency of a, pp.rance of B-body was 43.4${\pm}$1.3 percent in bull semen, and 45.5${\pm}$0.7 percent in hog semen. 3. A, pp.arance of B-body in bovine semen was increased due to duration of time after washing till 12 hours. 4. Separation of X- and Y- spermatozoa using diluents with different hydrogen ion concentration was impossible. 5. A, pp.arance of B-body separated in medium with 6, 10 and 20% ovalbumin was 51.1${\pm}$2.4, 50.6${\pm}$2.5 and 58.2${\pm}$3.0 percent, respectively, and those values were significiantly higher (p<0.01) than corresponding control values.

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