Spermatozoa viability can be assessed by microscopy, flow cytometry, and other methods using fluorescent stain. Flow cytometry can be used to examine the morphological and functional characteristics of spermatozoa in a short time. The purpose of this study was to compare the viability of cryopreserved spermatozoa in Jeju black cattle by two dual fluorescent stain methods. Semen of Jeju black cattle raised in Subtropical Livestock Research Institute, National Institute of Animal Science, RDA were collected with artificial vaginal technique. Sperm was diluted with Triladyl®-egg yolk diluent and then was performed cryopreservation.There was no significant difference in viability of spermatozoa according to the two dual fluorescent stain methods. However, when the distribution of spermatozoa according to the staining method was compared, the spermatozoa group stained with 6-CFDA/PI was more clearly distinguished than the spermatozoa group stained with calcein AM/PI.

본 연구는 섬진강 하구역에서 2017년 2월 대조기 만조와 간조, 3월 소조기 간조 동안 염분 구배에 따른 식물플랑크톤 생리활성도를 평 가하였다. 또한 2월 대조기 조사에서 나타난 특이적인 pH 변화특성을 파악하기 위해 추가적인 재현실험을 병행하였다. 현장조사 결과에 의하면, 만조와 간조에 수층 혼합에 의한 수직적인 염분의 차이는 미미하였지만, 담수역으로 향할수록 염분이 낮아져 수평적인 염분구배가 뚜렷하였다. 염 분과 영양염(질산염+아질산염: R=0.997, p<0.001; 규산염: R=0.979, p<0.001)과의 강한 음의 상관관계를 보였고, 이는 담수 기원의 질소와 규소의 높 은 농도의 영양염류가 해수와 혼합되는 과정에서 희석되고 있다는 것을 의미한다. 반면, 인산염농도는 염분이 15 psu를 기점으로 고염분의 유역에 서는 상대적으로 높은 농도가 관찰되었고, 이는 수심이 얕은 광양만의 퇴적물이 조석에 의한 수층혼합으로 재부유되어 인산염 재용출로 인해 수 층에 일정량 기여하였다는 것을 의미한다. 섬진강 하구역에서 동계 2월과 3월 우점한 식물플랑크톤은 Eucampia zodiacus로 나타났고, 그들은 전체 군집의 70% 이상을 차지하였다. 특히, 3월 생물 활성도를 평가한 결과, 염분과 chlorophyll a(R=0.82)뿐만 아니라 active chlorophyll a(R=0.80)과 강한 양의 상관관계를 보였다(p<0.001). 특히 담수의 영향을 강하게 받는 상류 정점에서는 해수종 규조류가 염분충격으로 생리활성을 잃어 최대 75% 사멸율을 기록하였다. 2월 대조기 조사에서는 상대적으로 형광값이 높은 정점에서 pH가 높게 나타났고, 이는 높은 밀도로 우점한 규조류의 광합성 작용으로 CO2가 다량 흡수되어 pH 상승효과를 가져온 것으로 판단되었다. 또한 2월 현장식물플랑크톤 군집을 농축하여 저염분 환경에서 생물농 도구배에 따른 pH 변화를 파악한 결과, 염분충격에 의해 사멸한 식물플랑크톤의 영향으로 광합성 활성도의 감소를 가져왔고, 우점 규조류의 사멸 로 인해 박테리아가 현저하게 증식하였다. 그 결과 박테리아가 규조류를 생분해하는 과정에서 CO2가 발생하여 pH 의 감소를 유발하였다. 결과적 으로 섬진강하구역에서는 염분 구배에 따른 생물의 광합성 활성도의 차이와 생물사멸이 뚜렷하게 관찰되었고, 이는 pH의 증감을 유발하는 중요한 인자로 파악되었다.

The present study was conducted to investigate the effect of BHT supplementation on sperm motility, viability, acrosomal integrity and plasma membrane integrity after frozen-thawing. One ejaculate was collected from one fertile Hanwoo bull by using artificial vagina at Hanwoo Research Institute. The ejaculate was transferred to laboratory immediately and diluted with pre-warmed semen extender (Optixcell, France) (1:1). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and divided into 5 groups according to BHT concentration (0, 0.5, 1.0, 2.0 and 4.0 mM) and cryopreserved in LN2 tank until evaluation. Frozen-thawed semen was transferred to 1.5 ml tube and incubated for 0, 2 and 4h. Sperm motility and motility parameters (total motility, VSL with 25μm≥, VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were evaluated by sperm class analysis (SCA, IVOS, Spain). There were not significant effects of BHT supplementation (0, 0.5, 1.0 and 2.0 mM) on total motile and VSL with 25μm≥ at 0, 2 and 4h. However, 4.0 mM of BHT supplementation showed negative effect on total motile (26.3%), VSL with 25μm≥ (1.3%) at 0 h (p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method and divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). There were no significant differences of LIA, LDA, DIA and DDA on various BHT concentrations at 0 and 2 h. However, 4.0 mM BHT supplementation showed decreased LIA compared with 0, 0.5, 1.0 and 2.0 mM BHT at 4 h (34.6, 37.1, 43.6, 45.4 and 14.7% vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.01). Addition of 4.0 mM of BHT showed negative effect on plasma membrane integrity compared with that of 0, 0.5, 1.0 and 2.0 BHT at 2 h (71.9, 64.2, 64.6, 67.5 and 31.7 % vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.05). In conclusion, various BHT concentrations on optixcell extender showed no improvement on sperm motility, viability and plasma membrane integrity.

Because of many benefits from raw seed sprouts, consumers have consumed them largely. However, despite of many benefits, raw sprouts has been implicated in food-borne diseases. The source of food-borne disease related to raw seed sprouts is thought to originate from seeds contaminated by pathogenic bacteria. So, Intense Pulsed Light (IPL), a non-thermal processing method, is an effective device for seeds to maintain microbial safety without loss of seed viability. The objective of this research was to determine the effects on microbial inactivation and quality in radish and wheat seed by IPL treatment and to figure out the correlation between inactivation of seeds and surface roughness value (Ra). At 5th day of germination, the average germination rate and shoot length of radish sprouts by IPL at total fluences of 121 J/cm2 were 95% and 5.8 cm. It was not significant compared to the control group. And log reductions of radish and wheat seeds by IPL showed 1.0 and 1.2, respectively. The results showed radish seeds have higher tolerance to IPL treatment than wheat seed. Also, radish seed had the rougher surface (Ra=2.85 μm) than wheat seed (Ra=0.55 μm). Therefore, IPL can decontaminate microbial population on seeds, but the effectiveness of IPL is dependent on the surface morphology of seeds.

The present study was aimed to determine the effect of green tea extract (GTE) and beta-mercaptoethanol (β-ME) supplementation in boar sperm freezing extender on sperm motility, viability and reactive oxygen species (ROS) level. Experimental groups were allocated into Lactose-egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/L GTE in LEY) and β-ME (50 μM β-ME in LEY). Spermatozoa extended with LEY were cooled to 5°C for 3 h and then kept at 5°C for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM (final sperm concentration: 1 × 108/mL). Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. Following thawing at 37°C for 25 sec, sperm viability and ROS level were measured using fluorescent double stain Fertility® and cytometry, respectively. Motility and viability of GTE supplemented-group were higher than those of control and β-ME without significance. ROS level in GTE group showed significantly lower than control (P < 0.05). In conclusion, GTE supplementation in boar sperm freezing extender can reduce ROS generation during freezing.

Cryopreservation of germ cells from genetically proven animals could be a source of restoration tools from the risk of extinction or disappearance of wanted characteristics. Using frozen semen, the genetic gains of Korean native cattle have been increased greatly for 70 years. The preservation of genetic resources as a form of frozen semen straw has limited availability due to the numbers. To circumvent this weakness of frozen semen, we tested two re-freezing methods with different initial thawing temperatures using frozen Korean proven semen and rare breed semen from albino, black and chikso breeders. It has been known that human sperm could resist to cryo-damages by repeated freeze-thaw cycles, but not for Korean proven bulls number (KPN) or for rare breeds. Total 7 frozen semem from brindled(2), black(1), Korean Albino(2) and KPN(1) bulls were used for our research. After thawing straws under 5°C/2min or 37°C/40sec with low temperature water bath and thermo jug, spermatozoa were re-diluted with triladyl diluents after first thawing and re-frozen. Sperm motilities were compared between animals and treated groups after re-thawing. Mean values of motility and viability of refrozen/thawed sperm for expansion of the number of straws were significantly higher in 5°C than in 37°C (P< < 0.05). However, the activity of viable sperm thawed at 5°C was significantly decreased after first and second thawing. It is estimated that re-freezing of frozen semen from rare Korean native cattle is possible with resistant properties of survived spermatozoa.

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation.To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and 1 μg/ml) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except 1 μg/ml (p<0.001).Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or 0.01 μg/ml AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and 0.1 μg/ml AFP (p<0.05), but increased with 1 μg/ml AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

Although cryopreservation of sperm is routinely used for clinical requirement, it has some problems, such as high generation of reactive oxygen species (ROS) and cold-shock. To reduce the detrimental damage in sperm, anti-oxidants were added to cryoprotectant for sperm. Curcumin is one of anti-oxidants, which are added in cryoprotectants. However, recent studies have demonstrated that curcumin decreases sperm viability and motility. This study was performed to identify the effect of curcumin on hydrogen peroxide (H2O2)-exposed bovine sperm, which were cryopreserved-thawed. In H2O2-exposed bovine sperm, reactive oxygen species (ROS) were significantly reduced by treatment with curcumin in a dose-dependent manner (p < 0.05). Among tested concentrations of curcumin (1 to 50 μM), 30 and 50 μM curcumin showed anti-oxidant effect on H2O2-induced ROS generation. On the other hand, combination of 30 or 50 μM curcumin with anti-oxidant H2O2 increased the percentage of apoptotic sperm compared to only H2O2 treatment. Sperm viability was also decreased in the combination of 30 or 50 μM curcumin with H2O2 as judged by FDA/PI staining. H2O2–induced decrease in sperm progressive motility was recovered by treatment with 1 μM curcumin. These results show that high concentration of curcumin has anti-oxidant effect, but it has also cytotoxic effect on bovine sperm. Sperm viability and motility might be more affected by cytotoxic signals of curcumin compared to antioxidant signals.

The purpose of this study was to examine the effects of taurine and vitamin E on ovarian granulosa cells damaged by bromopropane (BP) in pigs. We evaluated cell viability, plasma membrane integrity (PMI) and apoptotic morphological change in porcine ovarian granulosa cells. The cells were treated with 1-BP (0, 5.0, 10, and 50 μM), 2-BP (0, 5.0, 10, and 50 mM), taurine (0, 5.0, 10, and 25 mM), and vitamin E (0, 100, 200, and 400 μM) for 24 h. 10 μM 1-BP and 50 μM 2-BP inhibited viability and PMI, and induced apoptosis in porcine ovarian granulosa cells (p < 0.05). Cell viability and PMI were increased by taurine (10 and 25 mM) and vitamin E (100 and 200 μM), and apoptosis decreased (p < 0.05). Finally, the porcine ovarian granulosa cells were co-treated with BPs (10 μM), taurine (10 mM) and/or vitamin E (200 μM). Cell viability and PMI in the co-treated cells were increased, and apoptosis was decreased. In conclusion, taurine and vitamin E can improve cell viability and inhibition of apoptosis in porcine ovarian granulosa cells damaged by bromopropane.

본 연구는 USCG phase-II의 형식승인 기준인 자연상태 생물군집의 75 % 이상 유지하여야 하는 평가체계에 대비하여 자연생물군집 농축 및 선박평형수관리시스템(Ballast Water Management System, BWMS) 처리 전 후 생물사멸시험을 실시하였다. 자연 식물플랑크톤군집의 농축 조사는 중영양수계인 장목만과 부영양화수계의 마산만에서 동계에 수행하였다. 장목만과 마산만에서 1톤 기준으로 생물을 농축하였을 경우, 10-50 μm 크기 생물 현존량은 4.7 × 10⁴ cells mL-1과 0.8 × 10⁴ cells mL-1 이었고, 농축생물의 생존율은 90.4 %와 88.0 %로 각각 나타났다. 특히 장목만에서는 Skeletonema costatum-like species 같은 체인을 형성하는 소형 규조류가 극우점한 반면, 마산만에서는 <10 μm보다 작은 편모조류 및 체인을 형성하지 않는 대형 와편모조류(Akashiwo sanguinea, Heterocapsa triquetra)가 우점하였다. 이와 같은 우점종 세포크기의 차이로 장목만 농축효율이 마산만보다 높게 나타났다. BWMS 장비를 통과한 처리 당일 생물 사멸률은 장목만이 90.4 %로, 마산만의 93 %보다 약간 낮았고, 장목만에서 BWMS 처리 5일 경과 후, 대조군의 대상생물의 사멸률은 6.7 %로 나타났다. 처리군에서는 >99 %로 대부분 사멸되어, 시험생물로서의 적용 가능성을 확인할 수 있었다. 결과적으로 동계와 같이 해역내 생물량이 낮을 경우, 주간 8시간 수행한 네트의 생물농축만으로는 USCG Phase II의 형식승인 기준인 500톤 탱크에 1.0 × 10³ cells mL-1 이상으로 자연생물 개체수 밀도를 충족하기는 쉽지 않다는 것을 파악하였고, 이를 보완하기 위해서는 일정기간 자연생물을 대량 배양 및 채집할 수 있는 시스템 도입이 필요할 것으로 판단된다.

Egg SCP ( –47.2 ±1.0℃) of Red-spotted Apollo butterfly, Parnassius bremeri, was lower than larva(–35.0 ±0.9℃). The difference (12℃) between egg and larva was revealed by chorion through scanning electron microscope (COXEM EM-30, Korea). But considering of about 6 months’ pharate 1st instar in egg from June to November, function of egg seems not to be overwintering mechanism but rather estivation. In order to identify estivation function of chorion heat stress was carried out. Experiment was conducted on the 4 conditions such as high temperatures of 25, 35, 45 and 55 °C for 2 hour, then allowed them to recover at 25 °C for 1 hour. Total 65 samples 0f 31 eggs and 34 pharate 1st instar out of an egg were used and replicated 3 times. All pharate 1st instar in egg and pharate 1st instar out of egg survived at 25℃ and 35℃. In case of 45℃ survival rate of pharate 1st instar in an egg (91.6±3.5%) was significantly higher than those (37.8±9.4%) of pharate 1st instar out of an egg. All was dead at 55℃.

The purpose of this study was to examine the effects of taurine and vitamin E on sperm characteristics damaged by bromopropane (BP) in pig. We evaluated toxicity of BP on viability, membrane integrity and mitochondrial activity of spermatozoa. 1-BP (0, 2.5, 5.0, 10, and 50 μM), 2-BP (0, 2.5, 5.0, 10, and 50 μM), taurine (0, 5.0, 10, and 25 μM) and vitamin E (0, 50, 100, and 200 μM) were treated in fresh boar semen for 6 h. 10 and 50 μM of 1-BP and 2-BP inhibited sperm viability, membrane integrity and mitochondrial activity in fresh boar semen (P<0.05). 25 μM of taurine increased sperm viability and membrane integrity (P<0.05), 100 μM of vitamin E enhanced viability and mitochondrial activity of sperm (P<0.05). Finally, 10 μM of 1-BP and 2-BP was co-treated with taurine (25 μM) and vitamin E (100 μM) in the fresh boar semen. The co-treated samples did affected viability, membrane integrity and mitochondrial activity of sperm. In conclusion, taurine and vitamin E can improve and maintain sperm quality in fresh boar semen.

SCPs on larvae and eggs of red-spotted apollo butterfly, P. brimeri were measured according to the method of Kim and Kim (1977) with a thermocouple, BTM-4208SD (LT Lutron, Taipei, Taiwan), to detect the release of the latent heat of fusion as body water freezed. SCP of larvae during March goes below –27.4±1.7℃ and egg scp during November goes –47.2±1.0℃. In order to identify the reason of the difference(-17℃) between egg and larva we took photographed egg through scanning electron microscope (COXEM EM-30, Korea). Chorion of P. bremeri were 100.1㎛, Papilio machaon and Sericinus montela in same family was 10.8㎛, 5.5㎛ respectively. P. bremeri was 10 times, 20times thicker than another species within the same family .

The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG (81.1±15.1), 1,2-PROH (88.0±21.1) or the control (99.9±21.3) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO (14.7±1.3), EG (12.1±1.1), Glycerol (15.2±1.8), and 1,2-PROH (11.5±1.3) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.5±0.7, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

고로쇠나무의 낙하종자량과 종자활력의 연년변이와 그 원인을 밝혀보고자, 평창군 소재의 중왕산 천연활엽수림에서 종자트랩을 설치하여 낙하종자량과, 낙하종자의 활력을 6년(2009~2014)간 조사하였다. 2009년부터 2014년까지의 낙하종자량은 각각 55,700, 4,060, 60,900, 45,670, 403,450 ea/ha였으며, 2014년에는 310ea/ha로 평년에 비하여 매우 적은 양이었다. 2013년은 고로쇠나무 종자풍년(masting year)이었다고 판단된다. 종자활력 간 낙하종자량은 통계적 유의차가 인정되었다. 가장 많은 낙하종자는 피해 또는 부후된 종자이며 평균 47.88%였고, 다음으로 쭉정이 종자, 미발육 종자, 건전종자 순으로 낮아졌다. 건전종자는 6.7~31.0% 범위, 평균은 16.74%로 가장 낮았다. 종자풍년인 2013년에 건전종자는 평년(320~17,840ea/ha)보다 월등히 많은 125,070ea/ha이 숲바닥에 떨어졌다. 한편 피해 또는 부후종자의 비율이 38%로 가장 낮았으나, 종자립수는 153,310립/ha로 가장 많았다. 이러한 결과는 종자풍년에 천연갱신을 위한 충분한 건전종자가 공급됨을 암시하고, 포식자물림 가설(seed predator satiation hypothesis)을 지지한다고 사료된다. 연구가 미흡한 활엽수종의 종자생산에 대한 장기연구가 필요하다고 사료된다.

고로쇠나무의 결실량 연년 변이 및 결실에 영향하는 기상 요인을 파악하고자, 강원도 평창군 중왕산 지역의 천연활엽 수림에서 종자트랩을 설치하여 낙하종자량을 2009년부터 2014 까지 조사하였다. 연도별 낙하종자량과 몇 기상인자 와의 상관관계를 분석하였다. 2009년부터 2014년까지의 종자 낙하량은 각각 55,700, 4,060, 60,900, 45,670, 403,450 ea/ha였으며, 2014년에는 310ea/ha로 평년에 비하 여 매우 적은 양이었다. 2013년은 고로쇠나무 종자풍년 (masting year)이었다고 판단된다. 종자활력 간 종자립수는 고도의 통계적 유의차가 인정되었다. 가장 많은 낙하종자는 피해 또는 부후된 종자이며 평균 47.88%였고, 다음으로 쭉 정이 종자, 미발육 종자, 건전종자 순으로 낮아졌다. 건전종 자는 6.7~31.0% 범위, 평균은 16.74%로 가장 낮았다. 종자 풍년인 2013년에 건전종자는 평년(320~17,840립/ha)보다 월등히 많은 125,070립/ha이 숲바닥에 떨어졌다. 한편 피해 또는 부후종자의 비율이 38%로 가장 낮았으나, 종자립수는 153,310립/ha로 가장 많았다. 이러한 결과는 종자풍년에 천 연갱신을 위한 충분한 종자가 공급됨을 암시하고, seed predator satiation hypothesis를 지지한다고 사료된다. 활엽 수종의 종자생산에 대한 장기연구가 필요하다고 사료된다.

Purpose : Generally, tissue contrast enhancement patterns depend on the T1 relaxation. The T1 relaxation effect increases with the contrast agent concentration resulting in high contrast to noise ratio. Therefore, the aim of our study was to to evaluate improvement of contrast using gadobutrol in cardiac MRI viability scans. Materials and Methods : Two-hundred eighty-four patients who were administrated with gadoterate meglumine with the gadolinium concentration of 0.5mmol/mL(standard group) and 120 patients administrated with gadobutrol with the gadolinium concentration of 1.0mmol/mL were included in this study(study group). The total amount of gadolinium was the same as 0.1mmol/kg in both groups(0.2cc was injected in 0.5mmol/mL and 0.1cc was injected in 1mmol/mL contrast media). Signal intensities and noise of normal myocardium (NM) and the left ventricular cavity (LV) were measured. The ROIs were as follows (1) ROI of normal myocardium, (2) ROI of the left ventricle, (3) ROI of background. Then, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were compared between standard and study group. MRI scanning was performed using a 1.5T MRI scanner (Magnetom Avanto, Siemens ). The body coil was used as the transmitter, and a dedicated 32-channel cardiac coil was used as the receiver. The acquisition of MRI scanning was performed with the following protocol: short axis IR-true FISP (TR/ TE:800/3.42, 8mm thickness with 2mm interslicegap, FOV:150×100㎟, matrix size:256×192, flip angle:25 degrees). The Statistical significance of parametric data was determined using Student t-test. A two-sided P value of less than 0.05 was considered to indicate statistical significance. All statistical analysis was performed using the SPSS software package (version 18; SPSS, Chicago, IL). Results : Mean SNR of the study group was higher in the study group than standard group by 25.13% in (NM) and 30.74% in (LV). (mean difference -1.61, -22.26; standard deviation 0.66, 4.86; t value -2.449, -4.580; p<.05, p<.01). Mean CNR was also higher in the study group by 31.29%(mean difference -20.65; standard deviation 4.42; t value -4.675; p<.01). And, the difference of SNR and CNR between study group and standard group were statistically significant. Conclusions : Both SNR and CNR were higher using gadobutrol administration. Considering the involuntary movement of the heart, this study can be said innovative. This study has limitation because the patient population is restricted to patients with suspected heart disease. This study has significance as the usefulness and the diagnostic value of gadobutrol was proved proposing measures to increase the diagnostic value. This study suggests the increased the diagnostic value using gadobutrol for the cardiac delayed-enhancement study. and, Additional research is required for patients with various diseases.

생쥐 2세포기, 4세포기, 8세포기를 각 발생 단계에서 채취하여 동결 보호제를 첨가한 서로 다른 배양액에서 배양하고, 배아의 동결 보존 및 융해 시 급속 처리와 저속 처리 단계로 비교하여 이들 조건이 배아의 생존과 발현에 미치는 영향을 조사하여 다음의 결과를 얻었다. 동결 보호제로 처리하여 배양액을 달리한 경우, 급속 단계에서는 모든 배양액에서 비슷한 발생율을 보였고, 저속단계의 4세포기와 8세포기는 D-PBS에서 높은 발생율을 보였다(P＜0.05, P＜0.01). 배아의 발생 시기에 따른 동결 보존 후, 발생율은 2, 4, 8세포기로 넘어갈수록 발생율의 증가를 보여 8세포기에서 발생율이 가장 높았다(P＜0.01). 동결 보호제의 처리단계에 따른 발생율은 2세포기의 급속 단계에서는 유사하였으나, 4세포기와 8세포기는 저속단계에서 높은 발생율을 보였으며(P＜0.05), 특히 8세포기에서 가장 높았다(P＜0.01).

본 연구는 다양한 조건에서 장기 보존한 느티만가닥버섯 균주의 균사 생존력과 재배성능 검증을 수행하였다. 균사 생존력은 장기 보존된 균주를 9, 21,33, 45개월 단위로 해동하여 PDA 평판배지에 접종한 다음 1, 2, 3, 4, 5차 계대배양에 따른 균사 생장길이를 측정하여 검증하였다. 액체질소에 보존한 균사를 해동 직후 실시한 1차 계대배양에서는 동결보호제로써 Glycerol 10%를 단용으로 처리한 실험구에서균사생장이 저조한 결과를 나타내었으나, 5차 계대배양에서는 버섯 균사의 생장력이 다른 실험구와 동일한 수준으로 회복되어, 저장방법에 따른 균사의 생존력에는 큰 영향이 없는것으로 판단된다. 그러나 느티만가닥버섯을 액체 질소에 장기간 보존하고자 할 때는 동결보호제로 사용되는 Glycerol 10%에 Trehalose5%를 첨가하여 보존해야 균사의 생존력을 유지하는데 도움이 된다.장기 저장에 따른 느티만가닥버섯 균주의 재배 성능 시험에서는 4oC 사면배지에 보호제로써 mineraloil을 사용하는 것이 사면배지 자체로 보존하는 것에비해 수확량이 감소하는 결과를 나타내므로 장기 저장시에 mineral oil을 사용하는 것은 지양해야 할 것으로 판단된다.