• Title/Summary/Keyword: A23187

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Effect of $Ca^{2+}$ Concentration in Fusion Medium on the Fusion, Nuclear Morphology and Development of Bovine Somatic Cell Nuclear Transfer Embryos (세포 융합액 중의$Ca^{2+}$ 농도가 소 체세포 핵이식란의 융합, 핵형 및 체외발육에 미치는 영향)

  • 조재원;김정익;박춘근;양부근;정희태
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.33-39
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    • 2002
  • This study was conducted to investigate the effect of $Ca^{2+}$ concentration in fusion medium on the fusion, nuclear morphology and the development of bovine somatic cell nuclear transfer embryos. Bovine skin cells were transferred into an enucleated oocyte and fused with cytoplasm in the fusion medium containing with 0.05 to 1.0 mM Cacl$_2$. Nuclear transfer embryos were activated with a combination of A23187 and cycloheximide. Nuclear transfer embryos were fixed at 3 h after fusion or cultured for 7 ~8 days. Fusion rate was significantly (P<0.01) increased by increasing the $Ca^{2+}$ concentrations in the fusion medium from 0.05 mM (56.6%) to 0.5 mM (50.1%) and 1.0 mM (84.3%). More than 80% of reconstituted embryos underwent premature chromosome condensation (PCC) with 0.05, 0.1 mM CaCl$_2$, whereas 54.5% and 59.3% of embryos formed pronucleus (PN) directly without PCC in the 0.5 and 1.0 mM CaCl$_2$, groups. Blastocyst formation rates were significantly (P<0.05) different between 0.1 mM and 1.0 mM CaCl$_2$groups. From the present result, it is suggested that the elevated $Ca^{2+}$ concentrations in fusion medium can enhance the fusion and blastocyst formation rates of bovine nuclear transfer embryos.bryos.

Nuclear Remodeling and In Vitro Development of Bovine Oocytes Following Nuclear Transfer of Bovine Fetal Fibroblasts (태아 섬유아세포로 핵치환된 소 난자의 핵의 재구성과정과 체외 배발달)

  • Um, J. H.;S. J. Uhm;Kim, N-H;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.59-67
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    • 2000
  • This study was investigated the developmental potential of bovine embryos following nuclear transfer with bovine fetal fibroblasts (BFF). BFF were isolated from a male 45-day-old-fetus. Non-starved BFF labeled with MitoTracker were transferred into perivitelline space of enucleated oocytes. BFF-oocyte units were fused by electric pulse, and then fused oocytes were activated with calcium ionophore A23187 and subsequently 6-dimethylaminopurine (6-DMAP). The resulting zygotes were placed into CRlaa bovine embryo culture medium. Transfer of the nucleus into enucleated oocyte led to premature chromosome condensation, swelling and pronucleus formation. Remodeled oocytes were developed to the mitotic and 2-cell stage at 18 to 26 h after nuclear transfer. The incidence of in vitro development to the blastocyst stages was 21% of fused oocytes. Mitochondria of BFF eliminated rapidly and were not detected at 8 h after fusion. These results suggest that BFF can be successfully reprogrammed in enucleated bovine oocytes, and that reconstructed embryos can develop to the blastocyst stage.

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Samsoeum inhibits systemic anaphylaxis and release of histamine, cytokine in vivo and in vitro

  • Kim, Su-Jin;Kim, Na-Hyung;Moon, Phil-Dong;Myung, Noh-Yil;Kim, Min-Chol;Lee, Ki-Taek;Jo, Hyung-Mook;Kim, Na-Hyun;Rim, Hong-Kun;Seo, Min-Jun;Kim, Jin-Man;Lee, Seung-Eun;An, Nyeon-Hyung;Lee, Kang-Min;Lee, Si-Hyung;Park, Yun-Jum;Jeong, Hyun-Ja;Um, Jae-Young;Kim, Hyung-Min;Hong, Seung-Heon
    • Advances in Traditional Medicine
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    • v.9 no.2
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    • pp.115-127
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    • 2009
  • Samsoeum (SSE) is used in traditional oriental medicine for various medicinal purposes. However, the exact mechanism that accounts for the anti-allergy and anti-inflammatory effects of the SSE is still not fully understood. The aim of the present study is to elucidate whether and how SSE modulates the allergic reactions in vivo, and inflammatory reaction in vitro. In this study, we showed that SSE significantly decreased compound 48/80-induced systemic anaphylaxis, ear-swelling response, histamine release from preparation of rat peritoneal mast cells and anti-dinitropheny IgE-induced passive cutaneous reaction. Also, SSE inhibited the expression of inflammatory cytokine and cyclooxygenase-2 in PMA plus A23187-stimulated human mast cells (HMC-1). In addition, we showed that anti-inflammatory mechanism of SSE is through suppression of nuclear factor-${\kappa}B$ activation and $I{\kappa}B-{\alpha}$ phosphorylation/degradation in HMC-1. These results provided new insight into the pharmacological actions of SSE as a potential molecule for therapy of inflammatory allergic diseases.

Antioxidant and anti-inflammatory activities of hot water extracts of Ligularia fischeri (곰취 열수 추출물의 항산화 및 항염증 활성)

  • Nam, Ho-Seob;Jung, Ji-Wook;Kim, Do-Wan;Ha, Hyo-Cheol
    • Food Science and Preservation
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    • v.24 no.6
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    • pp.834-841
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    • 2017
  • This study was carried out to investigate the anti-oxidative and anti-inflammatory effects of hot water extracts of Ligularia fischeri cultivated in Youngyanggun. We obtained hot water extract (HWE) and cold water extract (CWE) from L. fischeri. The anti-oxidative activities of L. fischeri extracts were measured by 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. The anti-inflammatory effects of L. fischeri were evaluated in human mast cell line-1 (HMC-1) cells stimulated with phorbol-12-myristate-13-acetate plus A23187 (PMACI). The solid yields of HWE was 150% higher than CWE solid yield. Total polyphenol contents of HWE were $198.07{\pm}0.24mg/g$. The value of anti-oxidative activities of HWE were shown $IC_{50}$ $28.2{\pm}0.04ug/mL$. We showed that HWE significantly reduced the PMACI-induced the production of IL-6 (0.01-1 mg/mL), IL-8 (0.1-1 mg/mL), and $TNF-{\alpha}$ (0.01-1 mg/mL). These results indicate that the HWE of L. fischeri can be used as a functional material due to its antioxidant and anti-inflammatory activities.

Effects of the Ethyl Acetate Soluble Fractions of Sophorae Radix In Langendorff Hearts (고삼 Ethyl Acetate 소분획이 적출 심장에 미치는 영향)

  • Kwon Kang Beom;Park Gwan Ha;Kim Woo Kyung;Kim Eun Kyung;Kim Goo Hwan;Ko Kwang Hak;Choi Yeon Seong;Zhang Geun Gook;Rho Seong Il;Han Dong Won;Cha Suk;Park Dae Young;Ryu Do Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1309-1316
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    • 2004
  • We previously reported that the ethtyl acetate(EA) soluble fraction of Sophorae Radix(SR) water extract had the preventive effects against cardiovascular anaphylaxis elicited in experimental animals. In this study, we tested the anti-anaphylatic effects of the nine kinds of EA soluble subfractions in animal models such as Langendorff heart and anesthetized rats. These results were obtained as followed ; Among nine kinds of SR EA soluble subfractions, N10-16-2 and N10-16-9 fractions have an effects improving cardiovascular anaphylaxis in guinea pig Langendorff hearts. In passively anesthetized rats, N10-16-2 and N1 0-16-9 fractions of SR EA soluble subfractions have an effects improving cardiovascular anaphylaxis. N10-16-2 and N1 0-16-9 fractions of SR EA soluble subfractions inhibited the decrease of histamine release induced by compound 48180 and A-23187 in rat peritoneal mast cells. These results suggest that N10-16-2 and N10-16-9 fractions of SR EA soluble subfractions involve anti-anaphylactic molecules in cardiovascular system.

Biological Activity of Bamboo Salt

  • Shin, Hye-Young;Na, Ho-Jeong;Moon, Phil-Dong;Seo, Sang-Wan;Shin, Tae-Yong;Hong, Seoung-Heon;Lee, Ki-Nam;Park, Rae-Kil;Kim, Hyung-Min
    • Food Industry And Nutrition
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    • v.9 no.1
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    • pp.36-45
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    • 2004
  • Bamboo salt has been used for the purpose of prevention and treatment of various diseases in Korea. Present study was carried out to ascertain the effects of purple bamboo salt upon anti-allergic effect, anti-inflammatory activity and immune-enhance effect as well. Purple bamboo salt significantly inhibited the ear swelling response and histamine release induced by compound 48/80 in mice and rat peritoneal mast cells. Purple bamboo salt (0.01 ∼ lg/kg) also dose-dependently inhibited the passive cutaneous anaphylaxis by oral administration. Purple bamboo salt (1 mg/mL) in hibited phorbol 12-myristate 13-acetate plus calcium ionophore A23187-stimulated tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-1${\beta}$ and IL-6 secretion, by 67.04${\pm}$0.08%, 68.01${\pm}$1.85%, 69.48${\pm}$0.54%, respectively. In addition, purple bamboo salt inhibited the expression of TNF-${\alpha}$ mRNA in HMC-1 cells. Finally, we investigated the effect of purple bamboo salt in the forced swimming test (FST) and the change of purple bamboo salt-mediated cytokine production from MOLT-4 cells. At the 7th, immobility time was significantly decreased in the purple bamboo salt-administration group (35.4 ${\pm}$5.9 s for 1 g/kg) in comparison with the control group (93.2 ${\pm}$ 15.45). After FST, the content of glucose in the blood serum was increased and the levels of blood urea nitrogen, lactic dehydrogenase was decreased in purple bamboo salt-administration group. However, it had no effect on the elevation of CK and TP level. Purple bamboo salt (1 mg/mL) significantly increased the interferon (IFN)-${\gamma}$ and IL-2 level compared with media control (about 3.7-fold for IFN-${\gamma}$, about 3.5-fold for IL-2, p〈0.05) but did not affect the IL-4.

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Na-Ca Exchange in Sarcolemmal Vesicles Isolated from Cat Ileal Longitudinal Muscle (고양이 회장 종주근에서 Na-Ca 교환 기전의 특성에 관한 연구)

  • Woo, Jae-Suk;Suh, Duk-Joon;Kim, Yong-Keun;Lee, Sang-Ho
    • The Korean Journal of Physiology
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    • v.23 no.2
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    • pp.237-252
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    • 1989
  • Effect of a $Na^+$ gradient on $Ca^{2+}$ uptake was studied in isolated sarcolemmal vesicles of cat ileal longitudinal muscle. $Ca^{2+}$ uptake was markedly stimulated in the presence of an outwardly directed $Na^+$ gradient. External $Na^+$, monensin and A23187 abolished the $Na^+-dependent$ $Ca^{2+}$ uptake. Monovalent cations such as $K^+$, $Li^+$, $Rb^+$, $Cs^+$ and choline could not substitute for $Na^+$ in enhancement of $Ca^{2+}$ uptake. Divalent cations such as $Ba^{2+}$, $Sr^{2+}$, $Mn^{2+}$ and $Cd^{2+}$ but not $Mg^{2+}$ inhibited the $Na^+-dependent$ $Ca^{2+}$ uptake. Increase in external pH in the range of 6.0 to 8.0 stimulated the $Na^+-dependent$ $Ca^{2+}$ uptake. Amiloride inhibited the $Na^+-dependent$ $Ca^{2+}$ uptake at concentrations above 0.5 mM, whereas diltiazem or vanadate did not. The apparent Km of the $Na^+-dependent$ $Ca^{2+}$ uptake for $Ca^{2+}$ was 18.2 ${\mu}M$ and apparent Vmax was 689.7 pmole/mg protein/5 sec. Kinetic analysis of the $Na^+-dependent$ $Ca^{2+}$ uptake showed a noncompetitive interaction between internal $Na^+$ and external $Ca^{2+}$. The dependence of $Ca^{2+}$ uptake on internal $Na^+$ showed sigmoidal kinetics and Hill coefficient for internal $Na^+$ was 2.52. Inside positive membrane potential generated by imposing an inwardly directed $K^+$ gradient and valinomycin significantly stimulated the $Na^+-dependent$ $Ca^{2+}$ uptake. These results indicate that a $Na^+-Ca^{2+}$ exchange system exists in the sarcolemmal membranes isolated from cat ileal longitudinal muscle and it might operate as an electrogenic process.

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Anti-Atopic Effects of Mixed Extracts from Date Plum, Persimmon, and Mulberry Leaves (고욤잎, 감잎 및 뽕잎 복합추출물의 항아토피 효과)

  • Cho, Byoung Ok;Yin, Hong Hua;Shin, Jae Young;Fang, Chong Zhou;Chang, Che Denis;Jang, Seon Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.501-509
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    • 2016
  • The present study investigated the anti-atopic effects of mixed extracts from date plum, persimmon, and mulberry leaves (DPME) on atopic dermatitis (AD)-like skin lesions in hairless mice. The in vivo results demonstrated that DPME treatment significantly reduced the dermatitis clinical score and epidermal thickness in AD-like skin lesions. Histological analyses showed that DPME treatment strongly inhibited dermal infiltration of inflammatory cells and activity of mast cells in AD-like skin lesions. DPME treatment inhibited production of serum IgE and interluekin (IL)-4 in hairless mice with AD. Moreover, DPME treatment significantly suppressed production of tumor necrosis factor $(TNF)-{\alpha}$ and IL-6 cytokines in phorbol 12-myristate 13-acetate plus calcium ionophore A23187-stimulated HMC-1 human mast cells. In addition, DPME treatment reduced production of pro-inflammatory mediators (nitric oxide, prostaglandin E2, $TNF-{\alpha}$, and IL-6) in lipopolysaccharide-stimulated RAW 264.7 macrophages. Therefore, the results of this study indicate that the anti-atopic and anti-inflammatory effects of DPME may be involved in the regulation of inflammatory responses, suggesting that DPME may be used as an anti-atopic dermatitis material and natural anti-inflammatory ingredient.

Changes of Arachidonic Acid Metabolites in Silica-Exposed Alveolar Macrophage of Rats (유리규산분진에 폭로된 흰쥐의 폐포대식세포에 있어 아라키돈산 대사산물의 변화)

  • Lim, Young;Yun, Im-Goung
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.4
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    • pp.304-309
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    • 1992
  • Background: The alveolar macrophage may metabolize arachidonic acid through cyclooxygenase- and lipoxygenase- catalyzed pathways to produce a variety of metabolites of arachidonic acid. The production of these metabolites of arachidonic acid may enhance the defensive ability of the challenged lung. However, continued stimulation with the consequent production of proinflammtory metabolites of arachidonic acid, may ultimately enhance the disease process by contributing to chronic bronchoconstriction, fibrosis, and the persistent release of toxic oxygen species. Silicosis is an example of a disease process resulting from chronic exposure of the lung to foreign particles. This study was carried out to evaluate the changes of arachidonic acid metabolites from macrophages in experimental silicosis. Methods: We measured $PGE_2$, and $LTB_4$ in cultured macrophages taken from rats by radioimmunoassay at 24 and 48 hours after stimulation by silica dust, natural carbon dust, lipopolysaccharide, calcium ionophore (A23187) and medium (RPMI) as a control. For the experimental silicosis, 50 mg silica in 0.5 ml saline was administered intratracheally into the rat and grown to 20 weeks and measured $PGE_2$, and $LTB_4$ in the cultured macrophages lavaged from that rat. The used stimulants were the same as above. Results: 1) The amount of $PGE_2$ in the cultred macrophages from normal rat was significantly decreased in the group which was stimulated with silica dust for 48 hours compare with control non-stimulated group. 2) In the experimental silicosis group, $PGE_2$, release in cultured macrophages after 48 hours incubation with silica and natural carbon dust tended to be lower than those of non-stimulated group. 3) There were marked changes of $LTB_4$ in the groups of normal rats which were incubated with silica for 24, 48 hours and natural carbon for 48 hours compared with non-stimulated group. 4) In the experimental silicosis group, the release of $LTB_4$ was significantly increased macrophages cultured with silica and natural carbon dust after 24 and 48 hours incubation compared with non-stimulated group. Conclusion: The results of these studies suggest that the in vitro exposure of rat alveolar macrophge to silica and coal dust results in an alteration in alveolar macrophage metabolism of arachidonic acid that may promote an inflammatory reaction in lung tissue.

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