• Title/Summary/Keyword: Cell counting

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Study on the Protective Mechanism of Vitamin C in the SH-SY5Y Cell Death Induced by the Streptozotocin (스트렙토조토신으로 유도된 신경세포사멸에서 비타민 C의 보호 기전 연구)

  • Lee, Seung-Hee;Han, Kyung-Hoon;Kim, Hyun-Jun;Park, Kwang-Sung;Han, Sung-Hee;Kim, Jung-Hee;Heo, Jae-Hyeok
    • The Korean Journal of Food And Nutrition
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    • v.31 no.4
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    • pp.457-463
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    • 2018
  • In this study, we analyzed the protective effects of the vitamin C in the streptozotocin (STZ)-induced apoptosis using the SH-SY5Y, a neuroblastoma cell line. The cells were pretreated with the vitamin C ($100{\mu}g$) for 30 min, followed by the 24-hr treatment with the 2.5-mM STZ. The cell-viability assay using the Cell Counting Kit (CCK)-8 revealed the cell-survival rate increased by 15% following the vitamin-C pretreatment compared to the STZ-only treatment. Moreover, we conducted the western-blot analysis to determine the protective effect of the vitamin C regarding the apoptosis. Compared to those in the STZ-only-treatment group, the p-ERK and Bcl-2 expressions increased in the vitamin-C-pretreatment group, whereas the p-JNK and Bax expressions decreased. The vitamin-C pretreatment increased the expression of the SOD-1, an antioxidant enzyme, by more than 30%, indicating its protective role in the STZ-induced oxidative stress. Also, we found both the intrinsic- and extrinsic-pathway mechanisms of the STZ-induced apoptosis. The results of this study $s{\mu}ggest$ vitamin C may help prevent the neurodegenerative diseases.

Characteristics and osteogenic effect of zirconia porous scaffold coated with ${\beta}$-TCP/HA

  • Song, Young-Gyun;Cho, In-Ho
    • The Journal of Advanced Prosthodontics
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    • v.6 no.4
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    • pp.285-294
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    • 2014
  • PURPOSE. The purpose of this study was to evaluate the properties of a porous zirconia scaffold coated with bioactive materials and compare the in vitro cellular behavior of MC3T3-E1 preosteoblastic cells to titanium and zirconia disks and porous zirconia scaffolds. MATERIALS AND METHODS. Titanium and zirconia disks were prepared. A porous zirconia scaffold was fabricated with an open cell polyurethane disk foam template. The porous zirconia scaffolds were coated with ${\beta}$-TCP, HA and a compound of ${\beta}$-TCP and HA (BCP). The characteristics of the specimens were evaluated using scanning electron microscopy (SEM), energy dispersive x-ray spectrometer (EDX), and x-ray diffractometry (XRD). The dissolution tests were analyzed by an inductively coupled plasma spectrometer (ICP). The osteogenic effect of MC3T3-E1 cells was assessed via cell counting and reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS. The EDX profiles showed the substrate of zirconia, which was surrounded by the Ca-P layer. In the dissolution test, dissolved $Ca^{2+}$ ions were observed in the following decreasing order; ${\beta}$-TCP > BCP > HA (P<.05). In the cellular experiments, the cell proliferation on titanium disks appeared significantly lower in comparison to the other groups after 5 days (P<.05). The zirconia scaffolds had greater values than the zirconia disks (P<.05). The mRNA level of osteocalcin was highest on the non-coated zirconia scaffolds after 7 days. CONCLUSION. Zirconia had greater osteoblast cell activity than titanium. The interconnecting pores of the zirconia scaffolds showed enhanced proliferation and cell differentiation. The activity of osteoblast was more affected by microstructure than by coating materials.

Expression and Effects of JMJD2A Histone Demethylase in Endometrial Carcinoma

  • Wang, Hong-Li;Liu, Mei-Mei;Ma, Xin;Fang, Lei;Zhang, Zong-Feng;Song, Tie-Fang;Gao, Jia-Yin;Kuang, Ye;Jiang, Jing;Li, Lin;Wang, Yang-Yang;Li, Pei-Ling
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3051-3056
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    • 2014
  • Previous studies have demonstrated that JMJD2A is a potential oncogene and is overexpressed in human tumors. However, its role in the endometrial carcinoma remains largely unknown. In this study, we discovered that JMJD2A was overexpressed in endometrial carcinoma, using immunohistochemistry, quantitative realtime polymerase chain reaction, and western blotting. Downregulation of JMJD2A led to reduced endometrial carcinoma RL95-2 and ISK cell proliferation, invasion and metastasis as asessed with cell counting kit-8, cell migration and invasive assays. Collectively, our results support that JMJD2A is a promoter of endometrial carcinoma cell proliferation and survival, and is a potential novel drug target.

Evaluation of the effects of a low dose of Asiasari radix on stem cell morphology and proliferation

  • Park, Jun-Beom;Lee, Ji-Eun;Jin, Seong-Ho;Ko, Youngkyung;Jeong, Su-Hyeon
    • The Journal of Korean Medicine
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    • v.37 no.2
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    • pp.85-92
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    • 2016
  • Objectives: Asiasari radix (A. radix) is a traditional herb medicine that has been used as an analgesic, antitussive, or anti-allergic remedy. This study was performed to evaluate the effects of low concentration of the Asiasarum heterotropoides extract on the morphology and proliferation of the human mesenchymal stem cells derived from periodontal tissue. Methods: Stem cells derived from gingiva were grown in the presence of A. radix at final concentrations that ranged from 0.001 to $0.01{\mu}g/mL$. The morphology of the cells was viewed under an inverted microscope and the analysis of cell proliferation was performed by using Cell Counting Kit-8 (CCK-8) on days 1 and 3. Results: The control group showed fibroblast morphology. The shapes of the cells in 0.001 and $0.01{\mu}g/mL$ of A. radix were similar to that of the untreated control group. The cultures growing in the presence of A. radix at day 1 showed an increase in the CCK-8 value. The relative values of CCK-8 assays of 0.001 and $0.01{\mu}g/mL$ of A. radix are $130.6%{\pm}1.8%$ and $129.3%{\pm}1.5%$, respectively, when the CCK-8 result of the untreated control group at day 1 is considered 100% (P = 0.051). Conclusions: Within the limits of this study, low concentrations of A. radix seemed to increase the proliferation of the stem cells that were derived from the gingiva and did not have adverse effects on the morphology of the cells.

Changes of Chloroplast Number per Guard Cell pairs of Leaves by Ploidy Level in Nicotiana tabacum L. cv. BY-4 (담배 식물체[Nicotiana tabacum L. cv. BY-4]의 배수성에 따른 공변세포의 엽록체 수 변화)

  • 배창휴;이연희;양덕춘;민경수;김호일;이호연
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.179-184
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    • 2001
  • We evaluated a possibility of the use of chloroplast number per guard cell pairs as a measure for ploidy level in the different ploidy levels of tobacco plant (Nicotiana tabacum L. cv. BY-4) . The guard-cell chloroplast numbers of leaves of haploid plant were a half of wild-type plant. Furthermore, the number of chloroplast per guard cell pairs of the leaves of doubled-haploid plant increased in two times compared with that of haploid plant. In addition, the chloroplast number was not changed in the F$_1$ progenies. The change of the chloroplast number by leaf stage was not observed. The results indicate that there is a strong relationship between ploidy level (2x and 4x) and chloroplast number per guard cell pairs. This relationship was also, observed in both in vitro and pot cultured plants. It was determined that the measurement of chloroplast number in guard cells of leaf epidermis is simple to use and less labour intensive, and hence can be considered a practical alternative to the chromosome counting methods or flow cytometry in the tobacco plant.

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Gossypol Induces Apoptosis of Human Pancreatic Cancer Cells via CHOP/Endoplasmic Reticulum Stress Signaling Pathway

  • Lee, Soon;Hong, Eunmi;Jo, Eunbi;Kim, Z-Hun;Yim, Kyung June;Woo, Sung Hwan;Choi, Yong-Soo;Jang, Hyun-Jin
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.645-656
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    • 2022
  • Gossypol, a natural phenolic aldehyde present in cotton plants, was originally used as a means of contraception, but is currently being studied for its anti-proliferative and anti-metastatic effects on various cancers. However, the intracellular mechanism of action regarding the effects of gossypol on pancreatic cancer cells remains unclear. Here, we investigated the anti-cancer effects of gossypol on human pancreatic cancer cells (BxPC-3 and MIA PaCa-2). Cell counting kit-8 assays, annexin V/propidium iodide staining assays, and transmission electron microscopy showed that gossypol induced apoptotic cell death and apoptotic body formation in both cell lines. RNA sequencing analysis also showed that gossypol increased the mRNA levels of CCAAT/enhancer-binding protein homologous protein (CHOP) and activating transcription factor 3 (ATF3) in pancreatic cancer cell lines. In addition, gossypol facilitated the cleavage of caspase-3 via protein kinase RNA-like ER kinase (PERK), CHOP, and Bax/Bcl-2 upregulation in both cells, whereas the upregulation of ATF was limited to BxPC-3 cells. Finally, a three-dimensional culture experiment confirmed the successful suppression of cancer cell spheroids via gossypol treatment. Taken together, our data suggest that gossypol may trigger apoptosis in pancreatic cancer cells via the PERK-CHOP signaling pathway. These findings propose a promising therapeutic approach to pancreatic cancer treatment using gossypol.

The Effect of Irradiation and Epidermal Growth Factor on Cell Cycle and Apoptosis Induction in Human Epithelial Tumor Cell Lines (수 종의 상피기원 종양 세포주에서 방사선 조사와 표피성장인자 투여에 따른 세포 주기의 변화와 apoptosis 유발에 관한 연구)

  • Han Won-Jeong;Heo Min-Suk;Lee Sam-Sun;Choi Soon-Chul;Park Tae-Won
    • Imaging Science in Dentistry
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    • v.30 no.1
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    • pp.71-79
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    • 2000
  • Purpose : This study was aimed to evaluate the cell cycle arrest and apoptosis induction after irradiation and epidermal growth factor (EGF) treatment in three human epithelial tumor cell lines (A431, Siha, KB). Materials and Methods: Single irradiation of 2, 5 and 10 Gy was done on three cell lines with 5.38 Gy/min dose rate using Cs-137 irradiator at room temperature. Also, EGF of 10 ng/ml was added immediately after 10 Gy irradiation. Cell growth was evaluated by counting the living cell number using a hemocytometer at 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Cell cycle arrest and apoptosis induction were assayed with the flow cytometry at 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Results : Growth of irradiated three cell lines were inhibited in proportion to radiation dose. EGF treatment after irradiation showed various results according to cell lines. On all cell lines, G2 arrest was detected after 8 hours and maximized after 12 hours or 1 day. Amount of G2 arrest was positively dose dependent. However, EGF showed no significant change on G2 arrest. G2 arrest was recovered with time at 2 Gy and 5 Gy irradiation. However, at 10 Gy irradiation, G2 arrest was continued. Apoptosis was detected at 10 Gy irradiation. On EGF treated group after irradiation, A431 and Siha cell lines showed slightly increased apoptosis but there was no statistically significant difference. KB cell line showed no marked change of apoptosis induction. Conclusion : Irradiation effects on cell cycle arrest and apoptosis induction in three human epithelial tumor cell lines, however epidermal growth factor doesn't effect on.

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Effects of Taurine on Glutamate-induced Neurotoxicity and Interleukin-6 mRNA Expression in Astrocytes

  • Yang, Seong-Chil;Baek, Su-Yeon;Choe, In-Pyo;Lee, Chang-Jung
    • Animal cells and systems
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    • v.1 no.3
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    • pp.467-473
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    • 1997
  • Taurine (2-aminoethanesulfonic acid), one of bioactive amino acid in the mammalian brain, is known to exert inhibitory effects on neurons via GABA receptor. In the present study, we examined effects of taurine on glutamateinduced neurotoxicity on hippocampal neuron cell culture using cell counting method and lactate dehydrogenase (LDH) assay. After 10 d of culture, cells were stimulated with appropriate drugs. Only 43% of cultured neuronal cells survived at one day after stimulation with 500 uM L-glutamate for 10 min. Survival rate was enhanced by 82% in the presence of 10 mM taurine. LDH activity from the culture supernatant incubated with a combination of L-glutamate and taurine was less than half of that with L-glutamate alone. In the next series of experiments, interleukin-6 (IL-6) mRNA expression in cultured astrocytes was investigated using reverse tanscription-PCR (RT-PCR). IL-6 mRNA was detected in the astrocytes stimulated with L-glutamate in a dose-dependent manner, while not detected in the unstimulated control astrocytes. The expression of IL-6 mRNA caused by 10 mM glutamate was inhibited by taurine, but not by GABA. These findings demonstrated a neuroprotective action of taurine against glutamate-induced toxicity.

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Parallel I/O DRAM BIST for Easy Redundancy Cell Programming (Redundancy Cell Programming이 용이한 병렬 I/O DRAM BIST)

  • 유재희;하창우
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.39 no.12
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    • pp.1022-1032
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    • 2002
  • A multibit DRAM BIST methodology reducing redundancy programming overhead has been proposed. It is capable of counting and locating faulty bits simultaneously with the test. If DRAM cells are composed of n blocks generally, the proposed BIST can detect the state of no error, the location of faulty bit block if there is one error and the existence of errors in more than two blocks, which are n + 2 states totally, with only n comparators and an 3 state encoder. Based on the proposed BIST methodology, the testing scheme which can detect the number and locations of faulty bits with the errors in two or more blocks, can be easily implemented. Based on performance evaluation, the test and redundancy programming time of 64MEG DRAM with 8 blocks is reduced by 1/750 times with 0.115% circuit overhead.

Polyploidy and Acclimatization Rate of Broccoli Plane Derived from Anther Culture (葯培養 由來 녹색꽃양배추(Broccoli)의 幼植物 馴化 및 培數性 調査)

  • 이광식;박재복;안춘희;윤여중
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.315-318
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    • 1998
  • This study was conducted to clarify the acclimatization rate and ploidy level of anther culture-derived plants in broccoli. The acclimatization rate from 71.4 to 100% was obtained from 7 varieties in 2 years. It was possible to identify the ploidy of the Plants obtained through anther culture by measuring the number of chloroplast in the guard cell. The average numbers of chloroplasts per guard cell in haploid diploid, and tetraploid were 8.5, 13.5 and 18.5, respectively. The regenerated plants could be classified based on these results into 47.1-51.3% of haploids, 47.9∼51.7%, diploid, and 0.8∼l.2% of tetraploids.

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