• Title/Summary/Keyword: in vitro study

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The effect of safflower seed fraction extract on periodontal ligament fibroblast and MC3T3-E1 cell in vitro (홍화씨 분획 추출물이 치주인대 섬유아세포와 MC3T3-E1 세포에 미치는 영향)

  • Huh, Ji-Sun;Kang, Jung-Hwa;Yoo, Yun-Jung;Kim, Chang-Sung;Cho, Kyoo-Sung;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.833-846
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    • 2001
  • Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.

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Effect of Trehalose and Sugar-addition on the Survival Rates of Bovine IVM/IVF Embryos after Vitrification (Trehalose 와 당첨가가 우 체외 수정란의 초자화 동결에 미치는 영향)

  • 양부근;김준국;정희태;박춘근;김종복;김정익
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.307-317
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    • 1998
  • This study was to investigate the vitrification method for cryopreservation technique of bovine in vitro fertilization(IVF) embryos. The morphological appearance and viability following vitrification of IVF bovine blastocysts and expanded blastocysts were examined. Embryos obtained 6, 7, 8 or 9 days after IVF were vitrified in both 40% ethylene glycoI(EG) plus 0.3M trehalose and 20% polyvinylpyrrolidone(PVP) in DPBS(ETP, Exp. 1) and ETP solution added 0.375M dextrose (ETPD, Exp. 2). The viability of Days 6, 7, 8 and 9 vitrified /thawed embryos at 24∼48 h culture after thawing was 11.9%, 19.8%, 23.4% and 15.3% in ETP(Exp. 1), and 34.6%, 54.5%, 37.9% and 13.0% in ETPD(Exp. 2), respectively. The viability of vitrified embryos produced from the culture days after IVF did not differ in Exp. 1, but significantly differ in Exp. 2(P<0.05). The viability of blastocysts and expanded blastocysts significantly differed(P<0.05) in 15.2% and 23.3%(Exp. 1), and 25.0% and 45.8%(Exp. 2). The result of Exp. 1 was similar to that of Exp. 2 on the viability of embryo according to developmental stages, but ETP solution plus sugar(dextrose) was increased the viability of vitrified embryos. In Experiment 3, The viability of vitrified embryos was not different between 12% and 20% PVP concentrations in ETP solution according to culture days or developmental stages. To investigate the effect of addition of sugar, two type of carbohydrates and a mixture of cryoprotectants for vitrification on the survival of bovine IVF embryos, bovine Days 7 to 9 blastocysts and expanded blastocysts were cryopreserved in either 20% glycerol plus 20% EG, 0.375M sucrose and 0.375M dextrose (GESD, Exp. 4) or 20% glycerol plus 20% EG, 0.3M trehalose and 20% PVP(GETP, Exp. 5) in DPBS. Survival rates of Day 7, 8 and 9 embryos at 24∼48h culture after thawing were 71.4%, 94.6% and 40.5% in GESD, and 59.5%, 81.5% and 62.5% in GETP, respectively. Hatching rates of Day 7, 8 and 9 embryos after thawing were 28.6%, 35.1% and 16.2% in GESD, and 27.0%, 33.3% and 18.8% in GETP, respectively. These results indicates that a mixture of cryoprotectants(glycerol and EG) and addition of sugar can improve the survival rates of the bovine IVF embryos(Day 7 or 8) vitrified, and the expanded blastocyst embryos are more suitable for vitrification than early blastocysts stage.

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An experimental study of dynamic frictional resistance between orthodontic bracket and arch wire (교정용 브라켓과 강선 사이의 운동마찰저항력에 관한 실험적 연구)

  • Lee, Jae-Hwan;Lee, Ki-Soo
    • The korean journal of orthodontics
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    • v.31 no.4 s.87
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    • pp.467-477
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    • 2001
  • This investigation was designed to determine the effects of wire size, bracket width and the number of bracket on bracket-wire dynamic frictional resistance during simulating arch wire-guided tooth movement in vitro. For simulation of an arch wire-guided tooth movement, we simulated tooth, periodontal ligament and cancellous bone. Maxillary premolar and 1st molar were simulated as real sized resin teeth, the simulated resin teeth which its root was coated by polyether impression material which its elastic modulus is similar to periodontal ligament were embedded in steel housing with inlay wax which its elastic modulus is similar to cancellous bone. Stainless steel wires in four wire size (0.016, 0.018, $0.016\;{\times}\;0.022,\;0.019\;{\times}\;0.025$ inch) were examined with respect to three (stainless steel) bracket widths (2.4, 3.0, 4.3mm) and the number of medium bracket(one, two, three) included in the experimental assembly under dry condition. The wires were ligated into the brackets with elastomeric module. The results were as follows : 1. In all the brackets, frictional resistance increased with increase in wire size. But, statistically similar levels of frictional resistance were observed between 0.018 inch and $0.016\;{\times}\;0.022$ inch wires in narrow bracket and also between 0.016 inch and 0.018 inch wire in wide backet. 2. The frictional forces produced by 0.016 inch wire were statistically similar levels in all the brackets. In 0.018 inch round wire, wide bracket was associated with lower amounts of friction than both narrow and medium brackets. In $0.016\;{\times}\;0.022,\;0.019\;{\times}\;0.025$ inch rectangular wire, wide bracket produced target friction than both narrow and medium brackets. In all the wirer, narrow and medium bracket demonstrated no statistical difference in levels of frictional resistance. 3. Frictional resistance increased with increase In number of medium bracket. 0.016 inch round wire demonstrated the greatest increment in frictional resistance, followed by $0.019\;{\times}\;0.025,\;0.016\;{\times}\;0.022$ inch rectangular wire which were similar level in increment of frictional resistance, 0.018 inch wire demonstrated the least increment. The increments of frictional resistance were not constantly direct proportion to number of bracket.

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Studies on Utilization Survey and Forage Quality of Phragmites communis and Miscanthus sinensis as Native Grasses in Paju and Ansan District, 2010 (2010 파주, 안산지역 갈대, 억새 등 야초류의 이용실태와 사료가치 평가)

  • Seo, Sung;Kim, Won-Ho;Jung, Min-Woong;Park, Hyung-Soo;Shim, Jae-Jin;Park, Jin-Gil;Sung, Ha-Guyn;Kim, Jong-Duk;Lee, Joung-Kyong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.2
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    • pp.151-158
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    • 2011
  • This study was carried out in 2010 to investigate the utilization and nutritive value of native grasses, such as Phragmites communis and Miscanthus sinensis which might reduce the cost of feeding domestic cattle. The regions within the Civilian Control Line in Paju and Sihwa region in Ansan were surveyed. In Paju, the yearly silage production was 900 MT harvested from 90 ha (10 MT/ha). About 30~50% of these were sold out at the marketing price of 52,000~55,000 Won per roll (130~137.5 Won/kg). Regrown Miscanthus sinensis of 70 cm in length contained 9.6% of crude protein (CP), 82.4 of relative feed value (RFV), and 67.7% of in vitro dry matter digestibility (IVDMD). Regrown Phragmites communis of 70 cm in length contained 13.8% of CP, 84.3 of RFV, and 67.9% of IVDMD. As harvesting was delayed, the quality was decreased dramatically. In Ansan region, Phragmites communis plantation covers 100 ha. The silage production was 550 MT from the area of 50 ha (11 MT/ha) in 2009. And the marketing price was 50,000 Won per roll. But in 2010 only a small amount of grasses could have been baled due to frequent and much rainfall from spring to summer season. However, the good forage quality was observed from regrown Phragmites communis of 80 cm in length, 12.9% CP, 99.8 RFV and 66.6% IVDMD, while that of late matured grasses was very low, 2.2% CP, 52.9 RFV and 36.4% IVDMD, greatly lower than forage quality of rice straw. The quality of grasses at bloom stage of 150 cm in length was similar to that of rice straw, showing 4.5% CP, 59.9 RFV and 42.2% IVDMD. In conclusion, the overall quality of most native grasses in this survey were very poor. Therefore, we recommend that Phragmites communis and Miscanthus sinensis should be harvested during June or July to obtain better forage quality which is richer in forage values than rice straw. Production of high quality forage crops was also desirable for self sufficiency of forage.

EFFECTS OF GINSENG COMPONENTS ON RODENTICIDE VACOR-INDUCED DIABETES MELLITUS IN RATS (인삼성분이 살서제(Vacor)로 유발시킨 쥐의 당뇨에 미치는 영향)

  • Lee Min-wha;Lee Tai-hee;Ahn Bong-whan;Park Byung-ju;Yang Sung-yeul
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.83-88
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    • 1984
  • It is now well established that the rodenticide Vacor (N-3-pyridyl-mehtyl-N'-p-nitropheny-lurea) causes a hyperglycemia in human and rats. It is also reported that there are some components (DPG-3) in ginseng radix which cause hypoglycemic effect on alloxan diabetic mice. In the present study, attempts were made to demonstrate in Vacor-poisoned rats the hypo-glycemic activity of red ginseng component(RGC), which was extracted by Kimura's DPG-3 extraction procedure and found to be effective for lowering a hyperglycemia in alloxan-diabetic rats. Vacor in a dose of $LD_{50}$ (10mg/kg) produced a glucose intolerance with a paradoxical moderate increase in blood immunoreactive insulin and derangement in glucose metabolism of epididymal adipocytes in rats. Although RGC (20mg/kg, i.p.) did not exert any significant influence on a hyperglycemia induced by large lethal doses (25mg/kg) of Vacor ingestion, it improved the LDso Vacor-induced glucose intolerance and caused a further increase in blood insulin levels in Vacor-poisoned rats. The administration of RGC (20mg/kg, i.p.) normalized Vacor-induced depression of glucose metabolism and lipogenesis in the epididymal adipocytes with an improvement of reduced responses to insulin of adipocytes from Vacor-poisoned rats. These results suggest that some red ginsneng components contained in RGC fraction normalize the depressed peripheral glucose unitlization and insulin response and eventually lead to an improvement of abnormal glucose tolerance developed in rats poisoned with small doses of Vacor.

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CONFOCAL LASER SCANNING MICROSCOPIC MORPHOLOGY OF DENTIN-RESIN INTERFACE AND ITS RELATIONSHIP WITH SHEAR BOND STRENGTH (상아질-레진 계면의 공초점 현미경적 형태 및 전단결합강도와의 관계)

  • Choi, Nak-Won;Cho, Byeong-Hoon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.24 no.2
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    • pp.310-321
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    • 1999
  • In this in vitro study, confocal laser scanning microscopic morphology of dentin-resin interface and its relationship to shear bond strength were investigated after the exposed dentin surfaces were treated with 3 different kinds of dentin adhesive systems[three-step; Scotchbond Multi-Purpose Plus(SMPP), self-priming bonding resin; Single Bond(SB), self-etching primer; Clearfil Liner Bond 2(LB2)]. 52 extracted human molar teeth without caries and/or restorations. The experimental teeth were randomly divided into three groups of seventeen teeth each. In five teeth of each group, class V cavities(depth: 1.5mm) with 900 cavosurface angles were prepared at the cementoenamel junction on buccal and lingual surfaces. Bonding resins of each dentin adhesive system were mixed with rhodamine B. Primer of SMPP was mixed with fluorescein. In group 1. the exposed dentin was conditioned with etchant, applied with above primer and bonding resin of SMPP. In group 2, with etchant and self-priming bonding agent of SB. In group 3, with self-etching primer and bonding agent of LB2. After treatment with dentin adhesive systems, composite resin were applied and photocured. The experimental teeth were cut longitudinally through the center line of restoration and grounded so that about $90{\mu}m$-thick wafers of buccolingually orientated dentin were obtained. And, $70{\sim}80{\mu}m$-thick wafers sectioned horizontally, thus presenting a dentinal tubules at 900 to the cut surface of a remaining tooth, were obtained. Primer of SMPP mixed with rhodamine B was applied to these wafers. Confocal laser scanning microscopic investigations of these wafers were done within of 24 hours after treatment. To measure shear bond strength, the remaining twelve teeth of each group were grounded horizontally below the dentinoenamel junction, so that no enamel remained. After applying dentin adhesive systems on the dentin surface, composite was applied in the shape of cylinder. The cylinder was 5mm in diameter, and 2mm in thickness. Shear bond strength was measured using Instron with a cross-head speed of 0.5mm/min. It was concluded as follows ; 1. Hybrid layer of SMPP(mean: $4.56{\mu}m$) was thicker than that of any other groups. This value was not statistically significant thicker than that of SB(mean: $3.41{\mu}m$, p>0.05), and significant thicker than that of LB2(mean: $1.56{\mu}m$, p<0.05). There was a statistical difference between SB and LB2(p<0.05). 2. Although there were variations in the length of resin tag even in a sample, and in a group, most samples in SMPP and SB showed resin tags extending above $20{\mu}m$. But samples in LB2 showed resin tags of $10{\mu}m$ at best. 3. Besides primer's infiltration into demineralized peritubular dentin and dentinal tubules, fluorophore of primer was detected in the lateral branches of dentinal tubules. 4. All groups demonstrated statistically significant differences from one another(p<0.05), with shear bond strengths given in descending order as follows: SMPP(18.3MPa), SB(16.0MPa) and LB2(12.4MPa). 5. LB2 having thinnest hybrid layer($1.56{\mu}m$) showed the lowest shear bond strength(12.4MPa).

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Effect of Bulhwangeumjeonggi-san on Cytokine Levels of Mouse Th1/Th2 Cells and Anti-allergic Activity in Ovalbumin-sensitized Allergic Inflammation Model (불환금정기산(不換金正氣散)이 마우스 Th1/Th2 분화(分化) 및 알레르기 염증 반응 조절에 미치는 효과)

  • Lim, Kang-Min;Kang, Hee;Park, Sung-Min;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1467-1476
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    • 2006
  • This study was to evaluate the effect of Bulhwangeumjeonggi-san (BS) on mouse Th1 and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of BS extract were measured as well as the amount ${\beta}$-hexosaminidase in RBL-wH3 cells and the levels of TNF-${\alpha}$ and IL-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with BS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total lgE, OVA-specific lgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4- T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of BS extract, it increased proliferation of CD4 cells by 14% in 50 ${\mu}g/m{\ell}$ concentration but it showed an inhibition in higher concentrations. CD4 T cells under Th1/Th2 polarizing conditions for 3 days with BS resulted in mild decrease of IFN- g in Th1 cells and mild increase of IL-4 in Th2 cell at 50 ${\mu}g/m{\ell}$ but the level of IL-4 decreased by 18% at 100 ${\mu}g/m{\ell}$. BS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}$-hexosaminidase in RBL-2H3 cells. Treatment of BS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-${\alpha}$ production. Oral administration of BS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum lgE and OVA-specific lgE by 50% and 55%, respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 25% and significant decrease of IL-4 and IL-5 by 53%, and 38%, respectively. The results show that BS does not strongly induce mouse T cells to transform into Th1 or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

Microenvironments and Cellular Proliferation Affected by Oxygen Concentration in Non-Small Cell Lung Cancer Cell Line (비소세포폐암주에서 산소 농도에 따른 미세 배양 환경과 세포 증식능)

  • Shin, Jong Wook;Jeon, Eun Ju;Kwak, Hee Won;Song, Ju Han;Lee, Young Woo;Jeong, Jae Woo;Choi, Jae Cheol;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.242-250
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    • 2007
  • Background: Abnormal angiogenesis can induce hypoxia within a highly proliferating tumor mass, and these hypoxic conditions can in turn create clinical problems, such as resistance to chemotherapy. However, the mechanism by which hypoxia induces these changes has not yet been determined. Therefore, this study was conducted to determine how hypoxia induces changes in cell viability and extracellular microenvironments in an in vitro culture system using non-small cell lung cancer cells. Methods: The non-small cell lung cancer cell line, A549 was cultured in DMEM or RPMI-1640 media that contained fetal bovine serum. A decrease in the oxygen tension of the media that contained the culture was then induced in a hypoxia microchamber using a $CO_2-N_2$ gas mixture. A gas analysis and an MTT assay were then conducted. Results: (1) The decrease in oxygen tension was checked the anaerobic gas mixture for 30 min and then reoxygenation was induced by adding a 5% $CO_2-room$ air gas mixture to the chamber. (2) Purging with the anaerobic gas mixture was found to decrease the further oxygen tension of cell culture media. (3) The low oxygen tension resulted in a low pH, lactic acidosis and a decreased glucose concentration in the media. (4) The decrease in glucose concentration that was observed as a result of hypoxia was markedly different when different types of media were evaluated. (5) The decrease in oxygen tension inhibited proliferation of A549 cells. Conclusion: These data suggests that tumor hypoxia is associated with acidosis and hypoglycemia, which have been implicated in the development of resistance to chemotherapy and radiotherapy.

A Comparative Study of Dry Matter Yield and Nutritive Value of Autumn Sown Forage Crops in Daejeon Area (대전지역 추파 사료작물의 건물수량 및 사료가치 비교 연구)

  • Lee, Hyung-Suk;Lee, In-Duk
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.26 no.4
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    • pp.249-256
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    • 2006
  • The object of this experiment was to suggest the optimum autumn sown small forage crops as next forage crops harvested maize in Daejeon area. The field trials was conducted from 2002 to 2004 at Chungnam National University in order to evaluate the dry matter yield and qualities of the autumn sown small forage crops. The experiment was consisted of split plots (main plot : forage crops as rye, barley and wheat, and sub plot 4 harvest stages as boot, heading, milk and dough) design with three replications. The average DM yield for 2 years of rye was higher than that of other forage crops (p<0.01). As stages proceed, the DM yield tended to increase in all forage crops (p<0.01). In chemical composition, they had a difference on forage crops and stages. The crude protein (CP) was higher in barley (p<0.01), but fibrous contents such as NDF, ADF, cellulose and lignin were lower in wheat than in the other forage crops (p<0.01). In Daejeon area, although barley and wheat at milk and dough stages were observed with maximum yield and high quality as autumn sown forage crops, due to late harvesting periods, it is difficult to utilize them as previous forage crops seeded in maize. Based on the result mentioned above, it is concluded that rye harvested at boot to heading stages is the most suitable fur autumn sown small forage crops in Daejeon area. Meanwhile, in order to have the option of nutritional value in wheat and barley, they need to be supplied early maturing and high-yield variety of wheat and barley.

Role of Group II Phospholipase $A_2$ in the Pulmonary Oxidative Stress of the Acute Lung Injury Induced by Gut Ischemia-Reperfusion (장의 허혈-재관류로 유도된 급성 폐손상에서 산화성 스트레스에 관여하는 group II phospholipase $A_2$의 역할)

  • Jheon, Sang-Hoon;Kim, Keun;Lee, Sang-Cheol;Kim, Seong-Eun;Lee, Young-Man;Lee, Jong-Tae
    • Journal of Chest Surgery
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    • v.35 no.7
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    • pp.501-510
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    • 2002
  • Background: The various pathogeneses of acute respiratory distress syndrome have been suggested but not established yet. In the present study, the role of group II phospholipase $A_2$($PLA_2$) in the pathogenesis of gut ischemia-reperfusion(I/R) induced acute lung injury (ALI), especially in the pulmonary oxidative stress with infiltration of neutrophils was investigated. Material and Method: To induce ALI, reperfusion of mesentery was done for 120 min after clamping of superior mesenteric artery for 60 min in Sprague-Dawley rats that weighed about 300g. To exmaine the role of group II $PLA_2$ in ALI, especially endothelial injury associated with the action of neutrophils, lung myeloperoxidase activity, lung leak index, bronchoalveolar lavage fluid protein were measured, and pulmonary $PLA_2$ activity changes in gut I/R were also measured. The role of group II $PLA_2$in the neutrophilic generation of free radicals was assessed by inhibiting group II $PLA_2$ with rutin, manoalide and scalaradial. Furthermore, to verify the oxidative stress in the lung, histologic and free radical detecting cytochemical electron microscopy were done. Result: After reperfusion, ALI was developed with accumulation of neutrophils in the lung, which was confirmed by the increase of myeloperoxidase activity, lung leak index and bronchoalveolar lavage protein (p<0.001). The pulmonary and intestinal group II $PLA_2$ activities significantly increased after gut I/R which were reversed by rutin(p<0.001). In vitro, cytochrome-c reduction assay denoted the inhibitory effects of rutin, scalaradial and manoalide on the production of free radicals from isolated human neutrophils. Histologically, neutrophilic accumulation and pericapillary edema in the lung after gut I/R was detected by light microscopy which was suppressed by rutin. In $CeCl_3$ cytochemical electron microscopy, the increased production of hydrogen peroxide in the lung after gut I/R was confirmed and also the production of hydrogen peroxide was decreased by rutin. Conclusion: On the basis of these experimental results, the inhibition of group II $PLA_2$ seemed to mitigate gut I/R-induced ALI by suppressing the production of free radicals from the infiltrated neutrophils. Collectively, group II $PLA_2$ seems to play a crucial role in gut I/R-induced ALI by neutrophilic oxidative stress.