• Title/Summary/Keyword: C3H/10T1/ 2 cell

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Antioxidant Activity and Protective Effects of Cirsium japonicum against Damaged Mouse Liver Cell (BNL CL.2) (엉겅퀴의 항산화 활성 및 손상된 흰쥐 간세포(BNL CL.2)에 대한 간 보호 효과)

  • Kim, Seonjeong;Kang, Seungmi;Ko, Keonhee;Nam, Sanghae
    • Journal of Life Science
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    • v.27 no.4
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    • pp.442-449
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    • 2017
  • We analyzed the content of total phenolic and silymarin compounds of Cirsium japonicum (CJ), and its antioxidant activities and Liver protective effects were compared with those of Silybum marianum (SM). The total phenolic content in the aerial part ($97.22{\pm}5.51mg/g$) of CJ is higher than that in the underground part ($85.32{\pm}3.06mg/g$). The total silymarin content of CJ was 55.56% of SM, with the underground part ($0.47{\pm}0.03mg/g$) having higher content than the aerial part ($0.18{\pm}0.02mg/g$). The antioxidant activity of CJ was generally slightly lower than that of milk thistle, and the underground part of CJ generally had higher activity compared to the aerial part. When CJ extracts were processed at 1 mg/ml, DPPH activities were $83.76{\pm}0.60$ and $88.28{\pm}0.17%$, and FRAP activities were $77.63{\pm}0.70$ and $82.83{\pm}0.39%$ for extracts from aerial part and underground part, respectively. ABTS activities were $68.60{\pm}1.24$ and $63.41{\pm}0.57%$ for underground and aerial part respectively when extracts were processed at 0.1 mg/ml. The Liver protective effects of CJ were higher in the extracts from underground part compared to the aerial part, Liver cells were damaged by treating them with t-BHP, $H_2O_2$ and Ethanol, and then they were treated with 0.2 mg/ml CJ extracts. The survival rates of the damaged liver cells were $49.58{\pm}0.34$, $76.87{\pm}1.10$ and $71.73{\pm}0.58%$ respectively, which were higher than the cells not treated with extracts.

Sex Determination in Somatic and Embryonic Cells of the Pig by FISH and PCR (FISH와 PCR에 의한 돼지 체세포 및 배아세포의 성 판정)

  • Chung, Y.;Jeon, J.T.;Kim, K.D.;Lee, S.H.;Hong, K.C.
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.323-331
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    • 1996
  • Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.

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The Effects of Diltiazem and Pentoxifylline on Apoptosis of Irradiated Rat Salivary Gland (흰쥐 침샘의 방사선조사시 Apoptosis에 대한 Diltiazem과 Pentoxifylline의 효과)

  • Yang, Kwang-Mo;Suh, Hyun-Suk
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.367-375
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    • 1998
  • Purpose : Xerostomia is a complication met by almost all patients who have radiotherapy for cancers of head and neck. Many studies for prevention of xerostomia will be necessary. Radiation-induced acute response of salivary glands has been defined as interphase death or apoptosis. Increased intracellular calcium level have an important role in radiation-induced apoptosis. Calcium channel blocker may prevent radiation-induced apoptosis of salivary glands. This study was designed to evaluate the effectiveness of diltiazem known as calcium channel blocker and pentoxifylline with inhibition of inflammatory response on the apoptosis as an acute response of radiation in rat salivary glands. Materials and Methods : Sprague-Dawley rats with about body weight 200-250 g were divided into 5 study groups : control, radiation alone, diltiazem with radiation, pentoxifylline with radiation, and diltiazem and pentoxifylline with radiation. The diltiazen and pentoxifylline were injected intraperitoneally 20 mg/kg and 50 mg/kg, 30 and 20 mimute before irradiation. respectively. Irradiation was given with a 4 MV linear accelerator. The 1600 cGy of radiation was delivered in a single fraction through a single anterior portal encompassing the entire neck. After 24 h of irradiation, rats were sacrificed and parotid and submandibular glands were removed and stained with hematoxylin and eosin. The quantification of apoptosis was performed by microscopic examination of stained tissue sections at a magnification of 200X and the percentage of apoptotic cell was calculated. Results : On parotid glands, the percentage of apoptosis by radiation alone, diltiazem with radiation, pentoxifylline with radiation, and diltiazem and pentoxifylline with radiation were 1.72$\%$ (8.35/486), 0.64$\%$ (2.9/453), 0.23$\%$ (1.2/516), and 0.28$\%$ (1.1/399), respectively. The apoptosis was markedly reduced in the groups receiving drugs compared with groups receivinge, radiation alone (p<0.05). In serous cell of submandibular glands, the percentages of apoptosis of radiation alone, diltiazem with radiation, pentoxifylline with radiation, and diltiazem and pentoxifylline with radiation were 1.94$\%$ (l1/567), 0.34$\%$ (1.9/554), 0.28$\%$ (1.8/637), and 0.22$\%$ (1.3/601), respectively. In the mucus cell of submandibular glands, the percentages of apoptosis were 0.92$\%$ (5.1/552), 0.41$\%$ (2.5/612), 0.29$\%$ (1.3/455), and 0.18$\%$ (1.0/562), respectively. The apoptosis was markedly reduced in the serous glands (p<0.05), but there was no difference in development of apoptosis in each group of mucus gland. Conclusion : These results suggest that radiation-induced apoptosis of serous cells of salivary glands may be decreased by diltiazem and pentoxifylline administration.

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Effects of Activation Treatments and Culture Condition on In Vitro Development of Caprine In Vivo and In Vitro Oocytes (재래산양의 체내 및 체외유래 난자의 활성화 처리방법 및 배양조건이 단위발생란의 체외발달에 미치는 영향)

  • Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.181-185
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    • 2004
  • This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

Comparison of Fermentation Characteristics of Italian Ryegrass (Lolium multiflorum Lam.) and Guineagrass (Panicum maximum Jacq.) during the Early Stage of Ensiling

  • Shao, Tao;Zhang, Z.X.;Shimojo, M.;Wang, T.;Masuda, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1727-1734
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    • 2005
  • The fermentation characteristics and mono- and di-saccharides compositions during the early stage of ensiling were studied with a temperate grass, Italian ryegrass (Lolium multiflorum Lam.) and a tropical grass, guineagrass (Panicum maximum Jacq.). The laboratory silos were kept in the room set at 25$^{\circ}C$, and then were opened on 0.5, 1, 2, 3, 5 and 7 days (14 days in Italian ryegrass) after ensiling, respectively. The Italian ryegrass silage showed a fast and large pH decrease caused by a fast and large production of lactic acid during the first 5 days of ensiling and succeeded to achieve lactic acid type fermentation; high lactic acid/acetic acid and lactic acid content at the end of ensiling (14 days), low values of pH (3.74), acetic acid, ethanol and ammonia-N/total nitrogen, none or only small amounts of Butyric acid, valeric acid and propionic acid. The guineagrass silage showed a slow decrease in pH and a slow increase in lactic acid content during the full ensiling period, causing a high final pH value, low contents of lactic acid, acetic acid, total volatile fatty acids and total organic acids. In Italian ryegrass silage, mono- and di-saccharides compositions decreased largely within the initial 0.5 day (12 h) of ensiling. Sucrose disappeared rapidly within the initial 0.5 day of ensiling, but fructose and glucose contents showed an initial rise by the activity of enzymes in plant tissues, and then decreased gradually. On the other hand, the contents of monoand di-saccharides in guineagrass showed the largest decreases due mainly to plant respiration within the initial 0.5 day of ensiling, and no initial rises in fructose and glucose contents during the early stage of ensiling because of the absence of fructans which are hydrolyzed into fructose and glucose in temperate grasses. In both silages, the rate of reduction in mono- and di-saccharides compositions within the initial 5 days of ensiling was ranked in the order of glucose>fructose>sucrose, suggesting that glucose and fructose might be more favorably utilized than sucrose by microorganisms and glucose is the first fermentation substrate. It was concluded that the silage made from Italian ryegrass with high moisture content had a good fermentation quality owing to the dominance of lactic acid bacteria and active lactic acid fermentation during the initial stage of ensiling. These results can be explained by rapid plant sap liberation and the high activity of plant enzyme hydrolyzed fructans into fructose and glucose within the initial 2 days of ensiling, which stimulate the homofermentative lactic acid bacteria growth. In ensiling a temperate grass, the physical characteristics may ensure the rapid onset of fermentation phase, which results from the smaller losses of water-soluble carbohydrates during the initial stage of ensiling and providing sufficient water-soluble carbohydrates for lactic acid bacteria. The silage made from guineagrass with intermediate dry matter and high initial mono- and di-saccharides content was stable silage. This could be explained by the higher incorporation of air during the very early stage of ensiling and the restriction of cell breakdown and juice release due to the properties of a tropical grass with coarse porosity and stemmy structures. These physical characteristics delayed the onset of lactic acid bacteria fermentation phase by extending the phases of respiration and aerobic microorganisms activity, causing the higher loss of water-soluble carbohydrates and the shortage of lactic acid bacteria fermentation substrates.

Coagulation and Fibrinolysis in Exudative Pleural Effusions (삼출성 흉막액에서 응고 및 섬유소 용해계에 관한 연구)

  • Ryu, Jeong-Seon;Lee, Hong-Lyeol
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1214-1222
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    • 1998
  • Background : The intrapleural hypofibrinolysis is caused by mainly excessive concentration of pleural plasminogen activator inhibitor-1 antigen(PAI-1 Ag), which binds tissue type plasminogen activator. In pleural inflammation induced by sclerosing agents for pleurodesis, levels of pleural PAI-1 antigen increase in relation to decreasing D-dimer levels. It has been known that the pleural mesothelial cells have the capability of secreting PAI-1 Ag in response to inflammation in vivo. Therefore, we estimated whether pleural inflammation changes the balance between fibrinolytic and coagulative properties in exudative pleural effusions. Method : The thirty cases was included in our study. We determined the pleural levels of glucose, lactic dehydrogenase(LDH), pH and the counts of white blood cell(WBC), polymorpho leukocyte(PMN), lymphocyte as the parameters of pleural inflammation and cellular components of pleural fluid. The plasma level of fibrinogen in fluid and the neutrophil count in blood were determined. The levels of D-dimer, PAI-1 Ag and thrombinantithrombin III complex(TAT) were determined by ELISA(Behring, Marburg, Germany). Result : The causes of pleural effusion were as following : tuberculous in 14 cases, malignant in 10 cases and parapneumonic in 6 cases. The levels of pleural D-dimer, PAI-1 Ag and TAT was significantly higher than that of plasma(p<0..001). The severity of pleural inflammation did not correlated with pleural D-dimer, PAI-1 Ag, TAT and their plasma levels. But the level of pleural TAT correlated with pleural WBC and lymphocyte count. Conclusion : We found that the severity of pleural inflammations did not correlated with pleural D-dimer, PAI-1 Ag, TAT and the possibility of local production of PAI-1 antigen is present.

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