• Title/Summary/Keyword: membrane integrity

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Leakage of Seed Reserve Nutrient in Artificially Aged Pepper Seeds and Enhancement of Seed Vigor by Priming (노화처리된 고추종자의 저장성분 누출과 priming 처리가 종자활력 증진에 미치는 영향)

  • Kang Jum-Soon;Choi In-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.352-356
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    • 2006
  • Quantity of protein, amino acid, and sugar leaked from seeds was greater as the viability of seeds was dropped by the time elapsed of seed aging treatment. In the seeds with the artificial aging treatment for 20 days, 35.8 mg of protein was leaked on the 4th day after soaking, which was 6.9 times higher than that of control. Leakage of amino acid was also higher from low quality seeds treated with the aging treatment. In the seeds with the aging treatment for 20 days, $36.5{\mu}g$ fig of sugar was leaked on the 4th day after soaking, which was 2.8 times higher than that of control. The leakage of inorganic compound was higher from the low quality seeds, and leakage of total protein, amino acid, and sugar. According to .the quantity of leakage, water soluble compounds, which can be used for the assessment of seed quality without any destruction, were protein and potassium. Germination rate and percentage of seeds were dropped with the seed aging treatment, and the seed viability could be recovered by priming treatment. This phenomenon was very clear when the low quality seeds were germinated at low temperature.

Effect of EDTA on canine parthenote development during in vitro culture (개 parthenote in vitro culture시 EDTA 첨가에 의한 발달율 향상)

  • Jeong, Haeyun;Zhao, Minghui;No, Jin-Gu;Ullah, Imran;Lee, Whi-Cheul;Wi, Hayeon;Ock, Sun A;Hur, Tai-young;Woo, Jae-Seok;Im, Gi-sun;Kim, Jong-Gug;Lee, Seunghoon
    • Journal of Embryo Transfer
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    • v.33 no.3
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    • pp.139-147
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    • 2018
  • Somatic cell nuclear transfer (SCNT) is a useful biotechnological tool for animal cloning. Until now, SCNT has been inefficient, especially in dog. It is believed that an embryo developmental block in SCNT embryos is cause of low production efficiency. However, no studies have been performed on canines for embryo developmental block. In this study, we attempted to evaluate the beneficial role of EDTA in canine parthenogenic (PA) embryos development to overcome embryo developmental block. The PA embryos were divided into 0.01 mM EDTA treated and non-treated groups. Embryo developmental efficiency was measured by activating chemically parthenote. After EDTA induction, PA embryos were evaluated for embryonic development, Reactive Oxygen Species (ROS) activity, mitochondrial integrity, ATP production and genomic activation. The EDTA treated PA embryos showed significantly higher survival rate and improved cavity formation compared to non-treated. Furthermore, cytoplasmic ROS level was mitigated and mitochondrial membrane potential was found significantly higher in EDTA treated group followed by higher ATP production. Moreover, major embryonic genomic activation specific markers/factors were also elevated in EDTA treated group. Conclusively, we elucidated that EDTA showed substantially positive effect to overcome embryo developmental block in canine.

Effects of Antioxidants Treatment on the Cryopreservation of Human Hematopoietic Stem Cells (인간 조혈모 줄기세포의 냉동보존에 미치는 항산화제의 영향)

  • Kim, Eung-Bae;Hong, Soon-Gab;Do, Byung-Rok;Kim, Kyung-Suk;Lee, Joon-Yeong
    • Development and Reproduction
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    • v.12 no.1
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    • pp.67-76
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    • 2008
  • Oxidative damage resulted from reactive oxygen species (ROS) is one of the main causes for the decrease of the viability during in vitro culture and cryopreservation process. This experiment was performed to determine the effects of antioxidants on the human hematopoietic stem cell (HSC) during cryopreservation procedure. HSCs cultured in vitro with or without antioxidants were frozen and then examined for stem cell potential after thawing. The cell viability of thawed HSC was increased in $\alpha$-tocopherol and ascorbic acid treatment group compared to control group ($62.7{\pm}8.0%$) and it was higher in 150 uM $\alpha$-tocopherol treatment group ($70.5{\pm}7.0%$). No significant difference was observed in the membrane integrity in all groups. In auto-differentiation rate, no significant difference was appeared in all groups, but was lower in 150 uM $\alpha$-tocopherol ($7.3{\pm}2.6%$) compared to control group ($10.1{\pm}1.6%$). These results demonstrate that treatment of antioxidants improves the efficiency of cryopreservation for HSC and $\alpha$-tocopherol may be considered effective antioxidant for the protective effect on HSC.

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Effects of Reactive Oxygen Species on Sperm Function, Lipid Peroxidation and DNA Fragmentation in Bovine Spermatozoa (소 정자에 있어서 활성산소계가 정자 기능과 지방산화 및 DNA 절편화에 미치는 영향)

  • Ryu, Buom-Yong;Chung, Yung-Chai;Kim, Chang-Keun;Shin, Hyun-A;Han, Jung-Ho;Pang, Myung-Geol;Oh, Sun-Kyung;Kim, Seok-Hyun;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.105-115
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    • 2002
  • Objective : To evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase (XO) system on sperm function, the change of sperm characteristics, lipid peroxidation, and DNA fragmentation in bovine spermatozoa. Materials and Methods: ROS were produced using a combination of 1000 uM X and 50 mU/ml XO. The ROS scavengers: superoxide dismu tase (SOD) (200 U/ml) and catalase (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without ROS scavengers at $37^{circ}C$ under 5% $CO_2$ incubator. Sperm movement characteristics by CASA (computer-aided sperm analysis), HOST (hypoosmotic swelling test), Caionophore induced acrosome reaction, malondialdehyde formation for the analysis of lipid peroxidation, the percentage of DNA fragmentation using the method of TdT-mediated nick end labelling (TUNEL) by flow cytometry were determined after 2 hours incubation. Results: The action of ROS on bovine spermatozoa resulted in a decreased in capacity for sperm motility, Ca-ionophore induced acrosome reaction and membrane integrity, an increased in malondialdehyde formation and the percentage of sperm with DNA fragmentation. In the effects of antioxidant, catalase completely alleviated the toxic effects induced by the ROS in terms of sperm function and characteristics, however SOD exhibited no capacity to reduce the toxic effects. Conclusion: The ROS can induce significant damages to sperm functions and characteristics. The useful ROS scavengers can minimized the defects of sperm function and various damages of spermatozoa.

Foliage Contact Herbicidal Activity of Dehydrocostus lactone Derived from Saussurea lappa (목향(Saussurea lappa) 유래 Dehydrocostus lactone의 경엽 접촉 살초 활성)

  • Cho, Kwang-Min;An, Xue-Hua;Chon, Jae-Kwan;Kim, Hyo-Sun;Chun, Jae-Chul
    • Korean Journal of Weed Science
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    • v.30 no.4
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    • pp.421-428
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    • 2010
  • A foliage contact herbicidal substance was separated from ethyl ether fraction in n-hexane extract of Saussurea lappa roots and identified as dehydrocostus lactone [(3aS,6aR,9aR,9bS)-3,6,9-trimethylidene-3a,4,5,6a,7,8,9a,9b-octahydroazuleno[5,4-d]furan-2-one](DHCL). When DHCL at 4,000 ppm was foliage-applied to two grasses and two broadleaf plants, greater than 85% necrotic injury was obtained from large crabgrass, maize and soybean, whereas only about 40% necrotic injury appeared in black nightshade, indicating that DHCL has no gross morphological selectivity, but shows difference in contact response among the plant species tested. Conductivity in incubation medium of the leaf disks treated with DHCL increased as the incubation time continued. Relatively low contact injury in black nightshade as compared with the other three plant species tested was attributed to decrease in absorption of DHCL due to relatively high amount of cuticle. DHCL did not require light in the herbicidal action and there were no inhibitory effects on seed germination and cell elongation. Acetyl-CoA carboxylase activity was inhibited by 30% and 58% at $100\;{\mu}M$ and $1000\;{\mu}M$ DHCL, respectively. These results suggested that the herbicidal action of DHCL was related with inhibition of fatty acid synthesis which in turn caused to weaken cell membrane integrity.

Some Factors Affecting Freezing of Boar Semen in 5 ml Maxi-straws

  • Dai, J.J.;Wu, C.F.;Zhang, Defu;Yin, F.Z.;Zhang, T.Y.;Liu, D.;Wu, H.L.;Li, L.L.;Yang, S.T.;Wang, L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.507-515
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    • 2009
  • A series of experiments were conducted to determine the suitable freezing and thawing temperatures for the freezing of boar semen in 5 ml maxi-straws. The ultrastructure, in vitro fertilization (IVF) and artificial insemination (AI) of frozen-thawed semen were also be evaluated. The 5 cm freezing height gave the best results not only in post-thaw motility rate (54.00%), but also in normal acrosome morphology rate (NAR) (80.23%). There was no significant difference in the post-thaw motility between different thawing temperatures and corresponding thawing times (p>0.05); the group of $52^{\circ}C$ and 25 s gave the highest motility rate (45.00%). As a whole, not only from the motility but also the NAR, thawing at $42^{\circ}C$ was better than the other two treatments. In the freezing packages, 5 ml maxi-straw gave a little lower mobility (40%), viability rate (49.58%), plasma membrane integrity rate (53.91%) and NAR (52.65%) than the 0.25 ml straw, but there was no significant difference between the two straw volumes (p>0.05). The IVF capacity of frozen-thawed semen in this experiment was similar to fresh semen. From ultrastructure observation, the main damage to boar spermatozoa after freezing was seen in the acrosome, such as swelling and formation of vesicles. After AI in recipient Shanghai White sows, frozen-thawed semen from 5 ml maxi-straws and pellets produced 72.2% and 80% conception rate and 7.8 and 8 litter sizes, respectively, and there was no significant difference between the 5 ml maxi-straw and the pellet (p>0.05).

Effect of Thawing Temperature on Sperm Characteristics of Frozen Semen in Miniature Pig (미니 돼지 동결정액의 융해 온도가 정자성상에 미치는 영향)

  • Choi, Won-Cheol;Yang, Mi-Hye;Lee, Yong-Seung;Cheong, Hee-Tae;Yang, Boo-Keun;Lee, Dong-Seok;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.175-179
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    • 2007
  • The objective of this study was to investigate the effect of thawing temperature on the sperm viability and acrosomal morphology for semen storage of miniature pig by the 0.5ml straw method. In this present study, sperm viability (SYBR-14/PI staining), membrane integrity (Hypoosmotic Swelling Test), acrosome intactness, intensity and capacitation status (chlorotetracycline staining) in frozen miniature pig sperm were evaluated after thawing at 37, 50 and $70^{\circ}C$ for 5, 10 and 45 sec, respectively. Interestingly, the results indicated that sperm thawed at $70^{\circ}C$ for 5 sec significantly (p<0.05) increased sperm viability, but lower the percentage of AR (acrosome reacted spermatozoa) pattern compared to sperm thawed at $37^{\circ}C$ for 45 sec and $50^{\circ}C$ for 10 sec. In terms of thawing condition, high temperature for a short time using the 0.5ml straw was improved cryosurvival of miniature pig semen. Therefore, appropriate thawing method for cryopreservation of miniature pig is required for increasing post-thawing viability.

Functional Analysis of Fibroblastic Reticular Cells Derived from Mouse Lymph Node via Bidirectional Crosstalk with T Cells (T세포와 양방향 작용을 통한 마우스 림프절로부터 분리된 fibroblastic reticular cell의 기능적 분석)

  • Park, Sung Hee;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1199-1208
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    • 2013
  • Fibroblastic reticular cells (FRCs) form the structural backbone of the T zone provide a guidance path for immigrating T cells in the lymph node (LN). FRCs may contribute directly to developing T-cell biology in the LN and allow analyses of fundamental aspects of FRC biology related to T cells. FRCs inhibited T-cell apoptosis, and FRC culture supernatants strongly induced the expression of Bcl-xL in T cells against doxorubicin. Coculture of FRC and T cells resulted in rearrangements of the actin cytoskeleton, as well as global changes in the morphology of the FRCs. In addition, when cocultured, the T cells adhered to the FRC monolayer, and the membrane intercellular adhesion molecule (ICAM)-1 was slightly increased by day-dependent manner. In contrast, the expression of soluble ICAM-1 was dramatically increased in a day-dependent manner. Several chemokines, such as CCL5, CXCL1, CXCL5, CXCL16, CCL8, CXCL13, and ICAM-1, and MMPs were expressed in FRCs sensed by tumor necrosis factor (TNF) families. Nuclear factor kappa B ($NF{\kappa}B$)-RelA of the $NF{\kappa}B$ canonical pathway was translocated into FRC nuclear by $TNF{\alpha}$. In contrast, p52 proteolyzed from p100, a counterpart of RelB of the noncanonical $NF{\kappa}B$ pathway, accumulated in the peripheral FRC nucleus by agonistic anti-$LT{\beta}R$ antibody. In summary, we propose a model in which FRCs engage in bidirectional crosstalk to increase the efficiency of T-cell biology. This cooperative feedback loop may help to maintain tissue integrity and function during immune responses.

Effect of Water Soluble Extract of Lichens on Oxidative Phosphorylation and Level of Metabolite in Rat Liver (지의류(地衣類) 추출물을 섭취한 흰쥐 간의 산화적 인산화 과정 및 대사물 측정(II))

  • An, Mi-Jung;Suh, Jung-Soon;Lee, In-Ja;Cho, Sung-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.145-150
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    • 1985
  • This study was conducted to examine the pharmacological effect of water soluble extract of Lichens (Parmelia, Physcia and Cladonia species) on liver-damaged rat by $CCl_4$ injection. Rat livers were damaged acutely and chronically by one-time injection of $CCl_4$ just prior to five days of experimental period and continuous injections in every three days for eight weeks of experimental period, respectively. During each period the experimental group was fed Lichens extract(5.5 mg of dry wt/ml) instead of water given to the control group. For both acute and chronic liver damage, the experimental group showed higher oxidative activity of hepatic mitochondria measured by state 3 respiration, P/O ratio, respiratory control and ATP synthesized, compard to the control group. Serum glucose was slightly higher in the experimental group but liver glycogen showed no significant difference between experimental and control groups. In experimental group, liver glucose-6-phosphatase activity was increased during first two days after acute liver damage, but not significantly different from control group during chronic damage. Liver lactate, malate plus fumarte and glutamate tended to be higher in the experimental group, especially for chronic liver damaged rat. It is concluded that Lichens extract stimulate cytoplasmic and mitochondrial oxidative activities and the possible mechanism of the latter is supposed to involve the preservation of membrane integrity by certain component(s) of water-soluble Lichens extract.

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Quality Control Tests and Acceptance Criteria of Diagnostic Radiopharmaceuticals (진단용 방사성의약품의 품질관리시험 및 기준)

  • Park, Jun Young
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.1
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    • pp.1-10
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    • 2021
  • Radiopharmaceuticals are drugs that contain radioisotopes and are used in the diagnosis, treatment, or investigation of diseases. Radiopharmaceuticals must be manufactured in compliance with good manufacturing practice regulations and subjected to quality control before they are administered to patients to ensure the safety of the drug. Radiopharmaceuticals for administration to humans need to be sterile and pyrogen-free. Hence, sterility tests and membrane filter integrity tests are carried out to confirm the asepticity of the finished drug product, and a bacterial endotoxin test conducted to assess contamination, if any, by pyrogens. The physical appearance and the absence of foreign insoluble substances should be confirmed by a visual inspection. The chemical purity, residual solvents, and pH should be evaluated because residual by-products and impurities in the finished product can be harmful to patients. The half-life, radiochemical purity, radionuclidic purity, and strength need to be assessed by analyzing the radiation emitted from radiopharmaceuticals to verify that the radioisotope contents are properly labeled on pharmaceuticals. Radiopharmaceuticals always carry the risk of radiation exposure. Therefore, the time taken for quality control tests should be minimized and care should be taken to prevent radiation exposure during handling. This review discusses the quality control procedures and acceptance criteria for a diagnostic radiopharmaceutical.