• Title/Summary/Keyword: TdT

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A study of the [$Ca^{2+}$] and the Apoptosis of the KB Cell Lines after 10Gy Irradiation (방사선조사 후 유표피암종세포내 칼슘농도의 변화와 apoptosis 발현에 관한 연구)

  • Moon Je-Woon;Lee Sam-Sun;Heo Min-Suk;Choi Soon-Chul;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.29 no.1
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    • pp.105-117
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    • 1999
  • Purpose: Ionizing radiations have been reported as an apoptosis initiating stimulus in various cells and it has established that sustained elevations in [Ca/sup 2+/] can lead to DNA fragmentation by Ca/sup 2+/-dependent endonucleases, ultimately resulting in apoptotic cell death. The previous experiments have been reported by using primarily thymocytes and lymphocytes and the change of [Ca/sup 2+/] was measured only by minutes or hours respectively. We need to evaluate [Ca/sup 2+/] in both several minutes and hours after irradiation of radiation of radiation therapy and verify the apoptotic cells. Materials and Methods: We have measured [Ca/sup 2+/] in human gingival epitheloid cancer cell with 10Gy irradiation, at minutely intervals and hourly intervals using digitized video-intensified fluorescence microscopy and the fluorescent Ca/sup 2+/ indicator dye, fura-2. In order to find out that the transient rise in [Ca/sup 2+/] could induced apoptosis, cells were incubated for 1 hour at 37℃ with TdT enzyme, rinsed and resuspended containing fluorescence and observed under a confocal fluorescence microscope. MTT assay was done to determine cell activity and LDH assay was done to determine the amount of necrotic cells. Results: After irradiation, the transient and temporal increasing of [Ca/sup 2+/] in the KB cells was founded. Though, there was no change in the intracellular [Ca/sup 2+/] at 30 minutes and 2 hours after irradiation. We could detect of DNA fragmented cells at 4 hours after 10Gy irradiated cells. There were no significant differences between 4 hour, 1 day, 3 day cells. There were no significant differences in MTT and LDH assay between the irradiated group and the control group after 4 hours and 1 day. Though after 3 days there were differences in MTT and LDH assay between the irradiated group was significantly decreased than the control group, in LDH assay the number of necrotic cell death of the irradiated was higher than the control group. Conclusion: In KB cells there were incipient and temporal increasing of the [Ca/sup 2+/] with 10Gy irradiation and the apoptosis was founded from 4 hours later which was earlier than seeing of the change of the amount of the cellular ability and necrosis.

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The Cytotoxic Effect of Oral Wet Wipes on Gingival Cells (시판 중인 구강청결티슈의 세포 독성 관찰)

  • Jung, Im-hee;Park, Ji Hyeon;Lee, Min Kyeng;Hwang, Young Sun
    • Journal of dental hygiene science
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    • v.18 no.2
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    • pp.76-84
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    • 2018
  • Wet wipes are being increasingly used because of their convenience. Particularly, oral wet wipes are useful for regular cleaning of a baby's mouth after birth. Therefore, the consumption of oral wet wipes has increased over the past few years and a variety of products are commercially available. However, product information on safety is not sufficiently provided and still raises doubts regarding adverse effects. To confirm the safety of wet wipes as an oral hygiene item and provide information for their use, we investigated the cytotoxicity of oral wet wipes and verified the underlying mechanism. The anti-bacterial effect of oral wet wipes was analyzed using the disk diffusion method. The cytotoxic effects of oral wet wipes were observed based on morphological changes using microscopy and determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in gingival epithelial cells and gingival fibroblasts. Evaluation of apoptosis by oral wet wipes was explored using propidium iodide flow cytometric analysis and a terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) assay. Apoptosis-related molecules were also analyzed using western blotting. Five types of oral wet wipes were tested, and two products from Fisher-Price and Dr. Kennedy revealed strong cytotoxic effects on gingiva epithelial cells and gingiva fibroblasts, although they also showed intense anti-bacterial effects on oral bacteria. Cell cycle arrest in the G2/M phase and apoptosis were observed based on treatment of extracts from Fisher-Price and Dr. KENNEDY. Relatively high TUNEL levels, reduction of proliferating cell nuclear antigen and cyclin-dependent kinase 4 expression, and fragmentation of poly (ADP-ribose) polymerase were also elucidated. These results suggest that commercial oral wet wipes could exert cytotoxic influences on oral tissue, although there are anti-bacterial effects, and careful attention is required, especially for infants and toddlers.

Current and Future Trends of Accelerators and Antidegradants for the Tire Industry

  • Hong, Sung-W.
    • Elastomers and Composites
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    • v.34 no.2
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    • pp.156-176
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    • 1999
  • Rubber chemicals such as accelerators, antidegradants, vulcanizing agents, processing agents and retarders are very important to the production and protection of tires and rubber goods. The use of accelerators and antidegradants are evaluated in various tire components. This paper will focus on how to vulcanize tires economically and maintain the physical properties of each tire component without severe degradation due to oxygen, heat and ozone. Also, new non-nitrosoamine accelerators and non-staining antiozonants will be discussed. Lastly, the future requirements of antidegradants and accelerators in the tire industry will be reviewed. Tires have been vulcanized with Sulfenamides as primary accelerators and either Guamdine's or Thiurams as secondary accelerators to achieve proper properties at service conditions. However, interior components such as the carcass can be vulcanized with Thiazoles as a primary accelerator to cure faster than the external components. Using the combination of Sulfenamide with secondary accelerators in a tire tread compound and the combination of a Thiazole and Guanidine in a carcass compound will be presented with performance data. Uniroyal Chemical and another Rubber Chemical Manufacturer have developed, "Tetrabenzyl Thiuram Disulfide," (TBzTD) as a non-Nitrosoamine accelerator, which could replace Nitrosoamine generating Thiurams. This new accelerator has been evaluated in a tread compound as a secondary accelerator. Also, Flexsys has developed N-t-butyl-2-benzothiazole Sulfenamide (TBSI) as a non-Nitrosoamine accelerator which could replace 2-(Morpholinothio) -benzothiazole (MBS), a scorch delayed Sulfendamide accelerator. TBSI has been evaluated in a Natural Rubber (NR) belt skim compound vs. MBS. An optimum low rolling resistant cure system has been developed in a NR tread with Dithiomorpholine (DTDM). Also, future requirements for developing accelerators will be discussed such as the replacement of DTDM and other stable crosslink systems. Antidegradants are divided into two different types for use in tire compounds. Internal tire compounds such as apex, carcass, liner, wire breaker, cushion, base tread and bead compounds are protected by antioxidants against degradation from oxygen and heat due to mechanical shear. The external components such as sidewall, chafer and cap tread com-pounds are protected from ozone by antiozonants and waxes. Various kinds of staining and non-staining antioxidants have been evaluated in a tire carcass compound. Also, various para-phenylene diamine antiozonants have been evaluated in a tire sidewall compound to achieve the improved lifetime of the tire. New non-staining antiozonants such as 2, 4, 6-tris-(N-1, 4-dimethylpentyl-p-phenylene diamine) 1, 3, 5 Trizine (D-37) and un-saturated Acetal (AFS) will be discussed in the tire sidewall to achieve better appearance. The future requirements of antidegradants will be presented to improve tire performance such as durability, better appearance and longer lasting tires.

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Enhanced antidiabetic efficacy and safety of compound K/β-cyclodextrin inclusion complex in zebrafish

  • Nam, Youn Hee;Le, Hoa Thi;Rodriguez, Isabel;Kim, Eun Young;Kim, Keonwoo;Jeong, Seo Yule;Woo, Sang Ho;Lee, Yeong Ro;Castaneda, Rodrigo;Hong, Jineui;Ji, Min Gun;Kim, Ung-Jin;Hong, Bin Na;Kim, Tae Woo;Kang, Tong Ho
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.103-112
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    • 2017
  • Background: 20(S)-Protopanaxadiol 20-O-D-glucopyranoside, also called compound K (CK), exerts antidiabetic effects that are mediated by insulin secretion through adenosine triphosphate (ATP)-sensitive potassium ($K_{ATP}$) channels in pancreatic ${\beta}$-cells. However, the antidiabetic effects of CK may be limited because of its low bioavailability. Methods: In this study, we aimed to enhance the antidiabetic activity and lower the toxicity of CK by including it with ${\beta}$-cyclodextrin (CD) (CD-CK), and to determine whether the CD-CK compound enhanced pancreatic islet recovery, compared to CK alone, in an alloxan-induced diabetic zebrafish model. Furthermore, we confirmed the toxicity of CD-CK relative to CK alone by morphological changes, mitochondrial damage, and TdT-UTP nick end labeling (TUNEL) assays, and determined the ratio between the toxic and therapeutic dose for both compounds to verify the relative safety of CK and CD-CK. Results: The CD-CK conjugate ($EC_{50}=2.158{\mu}M$) enhanced the recovery of pancreatic islets, compared to CK alone ($EC_{50}=7.221{\mu}M$), as assessed in alloxan-induced diabetic zebrafish larvae. In addition, CD-CK ($LC_{50} =20.68{\mu}M$) was less toxic than CK alone ($LC_{50}=14.24{\mu}M$). The therapeutic index of CK and CD-CK was 1.98 and 9.58, respectively. Conclusion: The CD-CK inclusion complex enhanced the recovery of damaged pancreatic islets in diabetic zebrafish. The CD-CK inclusion complex has potential as an effective antidiabetic efficacy with lower toxicity.

Puerarin pretreatment attenuates cardiomyocyte apoptosis induced by coronary microembolization in rats by activating the PI3K/Akt/GSK-3β signaling pathway

  • Chen, Zhi-Qing;Zhou, You;Huang, Jun-Wen;Chen, Feng;Zheng, Jing;Li, Hao-Liang;Li, Tao;Li, Lang
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.2
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    • pp.147-157
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    • 2021
  • Coronary microembolization (CME) is associated with cardiomyocyte apoptosis and cardiac dysfunction. Puerarin confers protection against multiple cardiovascular diseases, but its effects and specific mechanisms on CME are not fully known. Hence, our study investigated whether puerarin pretreatment could alleviate cardiomyocyte apoptosis and improve cardiac function following CME. The molecular mechanism associated was also explored. A total of 48 Sprague-Dawley rats were randomly divided into CME, CME + Puerarin (CME + Pue), sham, and sham + Puerarin (sham + Pue) groups (with 12 rats per group). A CME model was established in CME and CME + Pue groups by injecting 42 ㎛ microspheres into the left ventricle of rats. Rats in the CME + Pue and sham + Pue groups were intraperitoneally injected with puerarin at 120 mg/kg daily for 7 days before operation. Cardiac function, myocardial histopathology, and cardiomyocyte apoptosis index were determined via cardiac ultrasound, hematoxylin-eosin (H&E) and hematoxylin-basic fuchsin-picric acid (HBFP) stainings, and TdT-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Western blotting was used to measure protein expression related to the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. We found that, puerarin significantly ameliorated cardiac dysfunction after CME, attenuated myocardial infarct size, and reduced myocardial apoptotic index. Besides, puerarin inhibited cardiomyocyte apoptosis, as revealed by decreased Bax and cleaved caspase-3, and up-regulated Bcl-2 and PI3K/Akt/GSK-3β pathway related proteins. Collectively, puerarin can inhibit cardiomyocyte apoptosis and thus attenuate myocardial injury caused by CME. Mechanistically, these effects may be achieved through activation of the PI3K/Akt/GSK-3β pathway.

Optical properties of $ZnIn_2Se$ and $ZnIn_2Se_4$:Co single crystals ($ZnIn_2Se_4$$ZnIn_2Se_4$:Co 단결정의 광학적 특성)

  • 최성휴;방태환;박복남
    • Journal of the Korean Vacuum Society
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    • v.6 no.2
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    • pp.129-135
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    • 1997
  • Undoped and Co-doped $ZnIn_2Se_4$ single crystals crystallized in the tetragonal space group 142m, with lattice constants a=5.748 $\AA$ and c=11.475 $\AA$, and a=5.567 $\AA$ and c=11.401 $\AA$. The optical absorption measured near the fundamental band edge showed that the optical energy band structure of these compounds had an indirect band gap, the direct and the indirect energy gaps of these compounds decreased as temperature changed from 10 to 300 K. The temperature coefficients of the direct energy gaps were found to be $\alpha=3.71\times10^{-4}$eV/K and $\beta$=519 K for $\alpha=3.71\times10^{-4}$eV/K and $\beta$=421K for $ZnIn_2Se_4$: Co. The temperature coefficients of the indirect energy gaps were also found to be $\alpha=2.31\times10^{-4}$ eV/K and $\beta$=285 K for $ZnIn_2Se_4$, and $\alpha=3.71\times10^{-4}$eV/K and $\beta$=609 K for $ZnIn_2Se_4$:Co, respectively. Six impurity optical absorption peaks due to cobalt are observed in $ZnIn_2Se_4$:Co single crystal. These impurity optical absorption peaks can be attibuted to the electronic transitions between the split energy levels of$CO^{2+}$ ions located at Td symmetry site of $ZnIn_2Se_4$ host lattice. The 1st order spin-orbit coupling constant ($\lambda$), Racah parameter (B), and crystal field parameter (Dq) ARE GIVEN AS -$243\textrm{cm}^{-1}, 587\textrm{cm}^{-1}, \;and\;327\textrm{cm}^{-1}$, respectively.

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Correlations between the Capacity of In Vitro Fertilization and the Assays of Sperm Function and Characteristics in Frozen-thawed Bovine Spermatozoa (소 동결-융해 정자에 있어서 체외수정능력과 정자 기능 및 성상 분석법간의 상관관계)

  • Ryu, B.Y.;Chung, Y.C.;Kim, C.K.;Shin, H.A.;Han, J.H.;Kim, S.H.;Moon, S.Y.;Kim, H.R.;Choi, H.
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.275-289
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    • 2002
  • The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.