The present study investigated the effects of follicle stimulating hormone (FSH) and human chorionic gonadotrophin (hCG) on the nuclear maturation of canine oocytes. Oocytes were recovered from mongrel female ovaries in various reproductive states; follicular, luteal or anestrous stage. Oocytes were cultured in serum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.5, 1.0 or 10 IU) or hCG (Exp. 2: 0, 0.5, 1.0 or 10 IU) or both (Exp. 3: 1 IU FSH + 1 IU hCG) for 72 hr to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with 1.9 μM. Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. More (p<0.05) oocytes matured to MII stage when follicular stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the control, 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (p<0.05) maturation rate to MII stage was observed in follicular stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG supplemented groups (0 to 1.5%). However, the combination of FSH and hCG did not improve the nuclear maturation rate of canine oocyte (2.4 %) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to MII stage.

Vitrification has been suggested to be an effective method for the cryopreservation of human ES cells. However, the efficiency of vitrification with different vehicles remains a matter of ongoing controversy. The objective of this study was to assess the efficiency of cryopreservation in human ES cells by vitrification using different vehicles. A human ES cell line and a variety of vehicles, including microdroplet (MD), openpulled straw (OPS) and electron microscopic grid (EMgrid), were employed in an attempt to assess vitrification efficiency. In order to evaluate the survivability and the undifferentiated state of the postvitrified human ES cells, we conducted alkaline phosphatase staining and characterization via both RTPCR and immunofluorescence assays. The survival rates of the postvitrified human ES cells using MD, OPS and EMgrid were determined to be 61.5%, 66.6% and 53.8%, respectively. There also exist significant differences between slowfreezing and vitrification (p<0.01). However, no significant differences were detected between the vehicle types. Finally, the pluripotency of human ES cells after thawing was verified by teratoma formation. Cryopreservation using vitrification is more effective than slowfreezing, and the efficiency of vehicles proved effective with regard to the preservation of human ES cells.

To study the signaling effect of insulin-like growth factor-Ⅰ(IGF-1), transgenic mice containing IGF-1 Receptor (IGF-1R) cDNA fused to metallothionein promoter were produced by DNA microinjection into the pronucleus of mouse zygote. Three founders were produced with transgenic mice containing IGF-1R gene. Transgenic mice lines contained approximately 4～20 copies of transgenes per cell and transmission of this gene into the progeny with Mendelian manner were determined. The founder mice were mated with normal mice to produce F1 mice and then F2 mice. Transmission rates of IGF-1R transgene in the progeny mice were 25～60% in F1 generation and 40～50% in F2 generation. The mRNA expression of IGF-1R transgene in liver was analyzed using RT-PCR for IGF-1R gene in liver. When body weights of transgenic pups were measured during 4, 10 and 14 weeks after birth, IGF-1R transgenic mice grew faster than non transgenic littermates. This study indicated that growth regulation by IGF-1 signaling through IGF-1R can be elucidated using IGF-1R transgenic mice.

The inhibition of cell recovery might be proceeded via either the damage of the mechanism of the recovery itself or via the formation of irreversible damage which could not be repaired at all. Both these processes may take place at the same time. Any of t

This study was to taken to demonstrate the effects of exogenous nitric oxide(NO) on hu rnan pu lp cell s ‘ In volvement of cyclic 3’, 5' -monophosphate(cGMP) in p버 paJ protection induced by herne oxygenase-l (J-lO-l) against NO-induced cytotoxicity , By use of Western blotting and cell viabi lity assay, we have examined the cytotoxicity and J-lO-l induction in pulp cells that were treated with NO donor ‘ S-nitroso-N-acetyl-D, L-penici 1 lamine(SNAP) , We have assessed wheathel' HQ--l contributes the cytoprotective effect against the cytotoxicity caused by NO, and inves tigated the l'elationship between HO-l and cGMP in the s ignaling pathway, SNAP decreased cell via bility but in creased HO-l expl'ession in a concentl'ation- and time一dependent manner in hurnan pu lp cells NO-induced cyto toxicity was inhibited in the presence of the hemin(inducer of HO-l) , whel'eas was en hanced in the pl'esence zinc protoporphyrin IX(ZnPP IX, HO-l inhibitor), thus Lhe NO-induced cytoLoxicity was cOl'related with HO- l expression. R‘ etreatment with a rnemhrane-permeable cGMP analog, 8-bromo-cGMP, restored cell death and enhanced the HO-l protein expression induced by SNAP, ln contrast‘ inhibition of guanylate cyclase by lI-l -[1,2,4] ox adiazole[ 4,3 口]quinoxalin-l-one(ODQ) pretreated pulp cells to 1 mM SNAP resulting in marked cytotoxicity , These findings , demonstrating a link between J-lO-l, regulated thl'ough the cGMP system and NO-induced cytotox.icity in huma띠 p버 p ceJls , suggesti ng a protective 1'ole of HO-l in pulp infl ammatory disease

Previously, we have reported a plant extract isolated from Lysimachia foenum gracum Herba as a new environment friendly biopesticide that has the mycelial growth inhibition effect on Magnaporthe oryzae, the pathogenic fungus of the rice blast disease. For the finding of additional biopesticide candidate, we tested the mycelial growth inhibitory effects about 700 species of plant extracts on PDA media. Among them, the extract of Anemarrhena asphodeloides showed prominent inhibitory effect of which IC50 was 139.7 ㎍/㎖. Mycelial radii of M. oryzae were measured on PDA medium containing the four organic solvent fractions isolated from total extract from A. asphodeloides. Ethyl acetate fraction showed the impressive inhibitory effect of IC50, 54.12 ㎍/㎖. In the subsequent rice field test for the total extract of A. asphodeloides, we obtained encouraging 62.0% control rate of rice blast disease without any phytotoxicity. It is almost equivalent to that of chemical pesticides implying the applicability of the extract as a new biopesticide. In further study, the analysis of active ingredients of the extract would be necessary for the development of a new biopesticide and for the verification of cellular mechanism by which the mycelial growth of M. oryzae inhibited.

고무화합물 형태로 구성된 조영제의 병에 Syringe Connector의 Spike를 연결 시 고무의 찢김 정도를 알아보고 찢김 및 분쇄로 인한 합성고무의 혼입 유무와 분쇄된 합성고무가 검출 시 분쇄물의 크기를 실험을 통해 알아보고자 하였다. 그 결과 찢김 정도의 경우 Syringe Connector의 끝과 최초 접촉하는 앞면이 약 3.14±0.04 ㎜로 뒷면 보다 많이 찢겼으며, 실험 대상인 10 병의 조영제에서 평균 7 개에서 15 개로 모두 분쇄물이 검출되었다. 검출된 분쇄물을 이용하여 크기를 측정한 결과 평균크기는 약 7.89±0.31 ㎛이었다. 향 후 다양한 실험 및 분석방법을 통한 추가실험과 더불어 흡인된 분쇄물 차단을 위한 미세 필터타입 자동주입장치의 개발이 필요하며, 분쇄물 유입 시 치명적 사고를 대비하여 관련기관의 관심 또한 필요할 것으로 사료된다.

본 연구는 LPS로 유도된 RAW 264.7 세포에서 다양한 약용식품으로 사용되고 있는 올리브 잎과 가지 추출물의 항염증 효과를 확인하였다. 올리브 잎과 가지 추출물은 각각 RAW 264.7 세포에 대하여 세포독성을 나타내지 않았고, LPS 자극에 의한 NO 및 PGE2 생성을 농도 의존적으로 억제했다. 또한, 올리브 추출물은 LPS 자극으로 분비된 TNF-α, IL-1β 및 IL-6의 전염증성 cytokine의 분비량을 억제하였으며, 특히 200 μg/mL 농도에서 올리브 가지 추출물이 잎 추출물 보다 IL-6를 억제하는 것으로 나타났다. 대표적인 염증 관련 신호 전달 경로 인자인 iNOS 및 COX-2의 발현을 검토한 결과 올리브 추출물은 iNOS의 발현을 농도의존적으로 현저히 감소시키는 것으로 관찰되었으나, 각각의 올리브 추출물이 COX-2 발현에는 영향을 미치지 않은 것으로 관찰되었다. 이와 같은 결과는 올리브 각 부위별 추출물은 모두 iNOS 및 NO 조절 경로를 조절하는 것으로 사료되나 iNOS 및 COX-2 단백질 발현은 병립적이지 않을 수 있음을 제시하고 있다. 본 연구 결과로 올리브 추출물이 독성과 부작용이 적은 항염증 효능을 가진 기능성 화장품 소재로써 개발 가능성이 있다고 사료된다.

본 연구는 완주에서 재배된 세계 주요 11개 적포도 품종으로 제조된 적포도주의 향기 성분을 headspace-solid phase microextraction 분석법으로 확인하였다. 향기성분은 총 75종이 확인되었다. 아로마화합물은 그들의 OAV 값에 의해 5 그룹으로 나뉘었다. 알콜, 알데하이드, 에스테르, C6 화합물이 11개 적포도주의 주요한 향기성분이었다. Isoamy alcohol 알콜과 phenylethyl 알콜은 11개 포도주에서 공통적으로 꽃향기, 달콤한 향을 나타내는데 중요한 물질이었다. Octanoic acid, ethyl ester, hexanoic acid ethylester은 모든 레드와인에서 과실향과 꽃향, 달콤한 향을 내는 중요한 성분이었다. 1-Hexanol은 모든 포도주에서 분석되었으나 풀향을 나타내는 향으로 나타났다. Chanceller, Malbec, marchel, Nsrsha, Pinot Meunier, Sangiovetto 포도주의 주요 향기성분은 과실 향인 것으로 나타났으며 Cabernet Franc, Cabernet Sauvignon, Sauvignon Vert 포도주의 주요 향기성분은 풀향인 것으로 조사되었다. 또한, MBA와 Narsha 포도주의 경우 꽃향이 주요 향인 것으로 조사되었다. 본 연구 결과를 바탕으로 적포도주용 품종을 육성할 때 선발기준으로 향기성분 분석을 활용할 수 있을 것으로 판단되었다.

The study compared the juice and wine odorants of the Cheongsoo grape cultivar with those of Chardonnay and Reisling wines. The volatile compounds of the three grape varieties were analyzed using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The most common volatile compounds in the juices from the three cultivars were terpenes, C13-norisoprenoids, etones, alcohols, and aldehydes. Terpenes were established as the most abundant group of volatiles in Cheongsoo grape juice, where as aldehydes predominated in Chardonnay and Riesling juices. Forty-two volatile compounds (acids, alcohols, esters, and others) were detected in the three white wines. The concentration of esters was about four times higher in Cheongsoo wine than in Chardonnay and Riesling wines. Five esters found in the Cheongsoo wine, namely, isoamyl acetate, ethyl butanoate, ethyl hexanoate, ethyl octanoate, and ethyl decanoate, exhibited high odor activity values (OAV) of >1. Furthermore, only Cheongsoo wine had a high OAV for isoamyl acetate odorant, which is associated with banana and sweet aromas. Therefore, the abundant and varied esters are believed to be key volatile fruity/sweet odorants in Cheongsoo wine.