• Title/Summary/Keyword: fluorescence microscope

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Adverse effect of IL-6 on the in vitro maturation of porcine oocytes

  • Yi, Young-Joo;Adikari, Adikari Arachchige Dilki Indrachapa;Moon, Seung-Tae;Heo, Jung-Min;Lee, Sang-Myeong
    • Korean Journal of Agricultural Science
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    • v.48 no.3
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    • pp.607-615
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    • 2021
  • Cytokines are protein mediators that possess the ability to assist cell-to-cell communication in immune system-related activities. In general, pathogen endotoxins activate the release of inflammatory mediators, and with time, there is an increase in the cytokine levels in the body. Interleukin (IL)-6 mediates the acute-phase inflammatory response, and elevated IL-6 levels have been reported in peritoneal fluids of women with pelvic inflammation and endometriosis, thereby associating it with oocyte quality and infertility. To overcome subfertility or infertility in humans and animals, the present study was done to examine the effect of recombinant IL-6 on porcine oocytes matured in vitro and subsequently to determine the fertilization rate and embryo development. Porcine oocytes were incubated with varying concentrations of IL-6 (0 - 2 ㎍·mL-1) for 44 h followed by in vitro fertilization and culturing of the oocytes. The oocytes or embryos were fixed with 3.7% paraformaldehyde (PFA) and stained with fluorescence dyes, and the meiotic spindle, chromosome organization, fertilization status and embryo development were subsequently assessed under a fluorescence microscope. We observed induction of an abnormal meiotic spindle alignment in the oocytes incubated with IL-6 compared to the control oocytes incubated without IL-6. Moreover, significantly decreased fertilization rates and embryo development were observed for oocytes incubated with IL-6 (p < 0.05). Thus, an increased IL-6 level during oocyte maturation could be associated with fertilization failure due to an aberrant chromosomal alignment and a disruption of the cortical granules. Taken together, our results indicate that successful assisted reproduction can be achieved by controlling the levels of inflammatory cytokines.

AN EXPERIMENTAL STUDY OF THE IRRADIATION EFFECTS ON RAT PAROTID GLAND (방사선조사가 백서 이하선에 미치는 영향에 관한 실험적 연구)

  • Lee Kyu Chan;Lee Sang Rae
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.16 no.1
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    • pp.49-58
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    • 1986
  • This study was designed to investigate the irradiation effects on the rat parotid gland, applied to the head and neck region. For this experiment, twenty-four rats, feeded under the even condition, were used as experimental animals. Twenty rats were used for experimental group and the rest were assigned to the control group. The experimental group was singly irradiated with 10Gray through Cobalt-60 radiotherapy device, Picker model 4M 60 (Field size; l2×5㎝, SSD; 50㎝, Depth; 1㎝). The experimental animals of both group were sacrificed each four animals in 2 days, 1week, 2weeks, 3weeks and 4weeks after irradiation. The specimens were examined through the light microscope using the H-E stain and H stain by routin procedure. The other specimens were observed under the fluorescence microscope using the B-O dichroic mirror and Y455 barrier filter after PA-ACH stain. 1. The results of this study were obtained as follows, The parotid acini were severely degenerated and the intraacinar spaces were widened. Within the acini, retained secretory granules and increased fibrosis were observed. Also the shape and the size of the acini showed very irregular atrophic degenerations. 2. The nuclei showed severe pyknosis, displacement and irregular aggregated appearance. 3. The tissue changes of the parotid acini were initiated after 2 days of irradiation and most severely appeared at the second week of irradiation, but almost returned to normal. 4. The salivary ducts of the parotid gland were severely atrophied, discontinued but initiated to regenerated after 3 weeks of irradiation.

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Study of On-chip Liquid Cooling in Relation to Micro-channel Design (마이크로 채널 디자인에 따른 온 칩 액체 냉각 연구)

  • Won, Yonghyun;Kim, Sungdong;Kim, Sarah Eunkyung
    • Journal of the Microelectronics and Packaging Society
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    • v.22 no.4
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    • pp.31-36
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    • 2015
  • The demand for multi-functionality, high density, high performance, and miniaturization of IC devices has caused the technology paradigm shift for electronic packaging. So, thermal management of new packaged chips becomes a bottleneck for the performance of next generation devices. Among various thermal solutions such as heat sink, heat spreader, TIM, thermoelectric cooler, etc. on-chip liquid cooling module was investigated in this study. Micro-channel was fabricated on Si wafer using a deep reactive ion etching, and 3 different micro-channel designs (straight MC, serpentine MC, zigzag MC) were formed to evalute the effectiveness of liquid cooling. At the heating temperature of $200^{\circ}C$ and coolant flow rate of 150ml/min, straight MC showed the high temperature differential of ${\sim}44^{\circ}C$ after liquid cooling. The shape of liquid flowing through micro-channel was observed by fluorescence microscope, and the temperarue differential of liquid cooling module was measuremd by IR microscope.

The Ultrastructural Characteristics of Preimplantation Embryonic Envelope in the Rat (흰쥐 초기발생에서 배아막의 미세구조적 특징)

  • 홍순갑;이준영
    • Development and Reproduction
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    • v.2 no.2
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    • pp.157-163
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    • 1998
  • Preimplantation embryos of the rat was examined by the morphological changes in the cortical granule envelope (CGE), blastomere surface, and zona pellucida (ZP) of embryo after cortical reaction. The ultrastructural characteristics and CGE of embryos were observed with the scanning electron microscope and fluorescence microscope. In the ultrastructural characteristic of embryo surface, surface microvilli were shortened and the CGE-like structure existed above microvilli in eight-cell embryo. Rough spongy surface and decreased network numbers were key characters of embryonic ZP compared to unfertilized oocyte. The CGE formed by cortical reaction existed in perivitelline space during embryo development but it was thin and locally distributed ill contrast to fertilized oocyte. The present results indicate that cortical reaction forms cortical granule envelope in perivitelline space and causes not only zona hardening, but also ultrastructural changes in ZP and cell membrane of preimplantation embryos.

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Study of Chip-level Liquid Cooling for High-heat-flux Devices (고열유속 소자를 위한 칩 레벨 액체 냉각 연구)

  • Park, Manseok;Kim, Sungdong;Kim, Sarah Eunkyung
    • Journal of the Microelectronics and Packaging Society
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    • v.22 no.2
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    • pp.27-31
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    • 2015
  • Thermal management becomes a key technology as the power density of high performance and high density devices increases. Conventional heat sink or TIM methods will be limited to resolve thermal problems of next-generation IC devices. Recently, to increase heat flux through high powered IC devices liquid cooling system has been actively studied. In this study a chip-level liquid cooling system with TSV and microchannel was fabricated on Si wafer using DRIE process and analyzed the cooling characteristics. Three different TSV shapes were fabricated and the effect of TSV shapes was analyzed. The shape of liquid flowing through microchannel was observed by fluorescence microscope. The temperature differential of liquid cooling system was measured by IR microscope from RT to $300^{\circ}C$.

Radix Tetrastigma Hemsleyani Flavone Induces Apoptosis in Human Lung Carcinoma A549 Cells by Modulating the MAPK Pathway

  • Zhong, Liang-Rui;Chen, Xian;Wei, Ke-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5983-5987
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    • 2013
  • Radix Tetrastigma Hemsleyani Flavone (RTHF) is widely used as a traditional herb for its detoxification and anti-inflammation activity. Recently, several studies have shown that RTHF can inhibit growth and induce apoptosis in human cancer cell lines. However, the mechanisms are not completely understood yet. In this study we investigated the potential effects of RTHF on growth and apoptosis in human lung adenocarcinoma A549 cells as well as its mechanisms. A549 cells were treated with RTHF at various concentrations for different times. In vitro the MTT assay showed that RTHF had obvious anti-proliferation effects on A549 cells in a dose- and time-dependent manner. Cell morphological changes observed by inverted microscope and Hoechst33258 methods were compared with apoptotic changes observed by fluorescence microscope. Cell apoptosis inspected by flow cytometry showed significant increase in the treatment group over the control group (P<0.01). Expression of apoptosis related Bax/Bcl-2, caspases and MAPK pathway proteins were detected by Western blotting. The results showed that RTHF up-regulated the Bax/Bcl-2 ratio and cle-caspase3/9, cle-PARP expression in a dose-dependent manner. Expression of p-p38 increased, p-ERK decreased significantly and that of p-JNK was little changed in the RTHF group when compared with the control group. These results suggest that RTHF might exert anti-growth and apoptosis activity against lung cancer A549 cells through activation of caspases and Bcl-2 family proteins and the MAPK pathway, therefore presenting as a promising therapeutic agent for the treatment of lung cancer.

Cytotoxic activity and probable apoptotic effect of Sph2, a sphigomyelinase hemolysin from Leptospira interrogans strain Lai

  • Zhang, Yi-xuan;Geng, Yan;Yang, Jun-wei;Guo, Xiao-kui;Zhao, Guo-ping
    • BMB Reports
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    • v.41 no.2
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    • pp.119-125
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    • 2008
  • Our previous work confirmed that Sph2/LA1029 was a sphigomyelinase-like hemolyisn of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai. Characteristics of both hemolytic and cytotoxic activities of Sph2 were reported in this paper. Sph2 was a heat-labile neutral hemolysin and had similar hemolytic behavior as the typical sphingomyelinase C of Staphylococcus aureus upon sheep erythrocytes. The cytotoxic activity of Sph2 was shown in mammalian cells such as BALB/C mouse lymphocytes and macrophages, as well as human L-02 liver cells. Transmission electron microscopic observation showed that the Sph2 treated BALB/C mouse lymphocytes were swollen and ruptured with membrane breakage. They also demonstrated condensed chromatin as a high-density area. Cytoskeleton changes were observed via fluorescence confocal microscope in Sph2 treated BALB/C mouse lymphocytes and macrophages, where both cytokine IL-$1{\beta}$ and IL-6 were induced. In addition, typical apoptotic morphological features were observed in Sph2 treated L-02 cells via transmission electron microscope and the percentage of apoptotic cells did increase after the Sph2 treatment detected by flow cytometry. Therefore, Sph2 was likely an apoptosis-inducing factor of human L-02 liver cells.

Metabolism of Calcium in the Oocyte Maturation of Rat (흰쥐의 난자성숙에 있어서의 칼슘의 대사)

  • Hong, Soon-Gab;Lee, Joon-Yeong
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.357-366
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    • 1996
  • The present experiments aimed to investigate the metabolism of calcium during oocyte maturation in rat. The concentration of free calcium and calmodulin in oocytes was measured respectively by using of fluo-3/AM and FITC with microscope fluorescence spectrometer. The ultrastructural localization of calcium precipitates in oocytes was observed with the transmission electron microscope. Cumulus-free immature oocytes(GV-oocyte) were cultured in vitro through 15 hours. The free calcium concentration in GV oocyte was $55.9{\pm}3.5nM$. In calcium-containing medium, the free calcium concentration was increased in germinal vesicle breakdown(GVBD) oocyte($64.2{\pm}7.3nM$). In normal medium after calcium chelator treatment ($10{\mu}M$ BAPTA/AM), the free calcium contents were slightly lower than those in control group. In calcium-free medium, the free calcium content was drastically increased in GVBD($72.7{\pm}3.4nM$) and metaphase I - anaphase I ($88.0{\pm}3.4nM$) oocyte. In maturation rate of oocytes, GVBD rate was high in control group($82.9{\pm}6.55%$) and calcium chelator treatment group($91.2{\pm}4.4%$), but in calcium-free medium group, it was low and then the oocyte was degenerated without polar body formation. Relative content of calmodulin in oocyte was significantly(P<0.001) increased in metaphase I - anaphase I than in GV and GVBD oocyte. The calcium precipitates were observed in mitochondria and cytoplasm of GV oocyte but that were not observed in mitochondria of GVBD and metaphase I - anaphase I oocyte. And then the calcium precipitates reappeared in mitochondria of metaphase II oocyte. The above results indicate that changes in free calcium and calmodulin concentration of oocyte occur according to the maturational stages and the extracellular calcium is required during oocyte maturation. Also change of calcium localization in oocyte occurs according to the maturational stages.

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Physical and Chemical Effects of Extracellular Matrix on the Growth of Cardiomyocytes (HL-1) (세포외 기질 물질의 물리·화학적 영향에 따른 심근세포(HL-1)의 성장 연구)

  • Hong, Yoon-Mi;Choi, Seong-Kyun;Kim, Sun-Min
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.11
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    • pp.1229-1235
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    • 2011
  • The extracellular matrix (ECM) is a key factor affecting cell growth and adhesion to the culture surface, and it is also important for maintaining the innate characteristics of cells. Here, we describe the effects of the ECM on cardiomyocyte (HL-1 cell line) growth, viability, phenotype, and contractile ability. Five different ECM materials were investigated to analyze their effects on the cell growth. The physical morphology of the ECM-coated surfaces was scanned with an atomic force microscope (AFM), and the attachment, growth, proliferation, viability, and phenotype of the cells were analyzed using fluorescence immunostaining and an inverted phase contrast microscope.

A STUDY OF CHIDED TISSUE REGENERATION FOR IMMEDIATE IMPLANTATION WITH/WITHOUT HA AUGMENTATION : A STUDY IN DOGS (성견에서 발치 직후 Titanium plasma sprayed IMZ 임프란트 이식시 조직유도 재생술에 따른 골 재생력에 관한 연구)

  • Hwang Hie-Seong;Chung Moon-Kyu
    • The Journal of Korean Academy of Prosthodontics
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    • v.30 no.3
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    • pp.361-378
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    • 1992
  • The purpose of this investigation was to evaluate the effect of the porous hydroxyapatite particles (Interpore $200^{(R)}$) and guided tissue regeneration membrane ($Gore-Tex^{TM}$ augmentation material) on amount and shape of generating new bone adjacent to implant. Implants were placed immediately after extraction in the bilateral 3rd, 4th premolars of the mandible of the adult dogs. In all experimental groups, artificial bony defects were formed at the buccal cortex area, 3.3mm in width and 3.0mm in depth. In the control group : sutured without HA particles & membranes after placing implants, the experimental group 1 : membrane was place over the artificial bony defect, the experimental group 2 : bony defect was filled with HA particles and covered with membrane. The examination of bone-implant interfaces using light microscope and fluorescent microscope concluded as follows. 1. In all three experimental groups, osseointegration was observed without epithelial migration. 2. In the healing degree of bony defect area, the experimental group 1, 2 showed more prominent healing than control group, and the experimental group 1 showed the most excellent bone formation. 3. In fluorescent microscopic finding, bone remodeling was observed in regenerated bone tissue at defect area of experimental group 1, but in experimental group 2, irregular, discontinuous linear fluorescence was observed at the lower portion of defect area and sign of bone remodeling was weak.

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