Amniotic membrane stem cells are considered as a good alternative to embryonic stem cells, but their use in clinical studies is still not common. Here, exosomes from canine amniotic membrane mesenchymal stem cells (cAmMSCexo) were used for dog sperm cryopreservation. Upon cryopreserved straws using cryoprotectant containing 0, 0.5, 1, or 2 μg/mL of cAmMSC-exo were thawed, motility and membrane integrity were analyzed. However, results showed no significant differences between the groups. We concluded that cAmMSC-exo with lower than 2 µg/mL have no effects on sperm cryopreservation, and further studies to get higher concentrations of cAmMSC-exo should be conducted for clinical application.

본 연구에서는 CFRP 복합재료 플레이트로 외부 보강된 철근콘크리트 보 구조물의 장기 거동 특성과 CFRP 복합재료 플레이트의 부착거동을 실험을 통하여 검토하였다. 장기 사용성을 고려하기 위하여 동결융해 시험법을 사용하였으며 4절점 휨 실험을 통하여 평가하였다. CFRP 복합재료 플레이트로 보강 후 동결융해를 진행한 시험체(FFCB)와 동결융해 후 CFRP 복합재료 플레이트로 보강한 시험체(LFCB)의 4절점 휨 실험 결과를 비교 분석하였다. LFCB 시험체가 FFCB 시험체 보다 조기에 CFRP 복합재료 플레이트의 박리가 발생하였으며, 파괴형태가 다르게 발생하였다. FFCB의 경우 콘크리트 커버가 탈락되었으며, LFCB의 경우 콘크리트의 표면의 탈락이 발생하였다. 동결융해의 영향을 받은 콘크리트 재료의 성질이 약화된 결과로 판단된다. 동결융해 실험을 통하여 콘크리트 공시체의 압축강도가 약 19% 감소하였으며. 이에 따라 콘크리트 커버부의 인장강도도 감소한 것으로 판단된다. 콘크리트 구조물에 대한 보강의 경우사용기간 및 환경변수 등을 고려하여, 콘크리트 보강표면에 대한 정확한 상태평가가 필요할 것으로 판단된다.

This study investigated the quality characteristics of 5 different types of food (beef tenderloin, Japanese Spanish mackerel, radish, blueberry and tofu) by applying liquid immersion freezing, which is one of the quick-freezing methods. Each sample was refrigerated by -20℃ conventional freezing or -70℃ liquid immersion freezing respectively. After thawing at 4℃, we examined drip loss, color value, hardness, and volatile basic nitrogen. The drip loss of -70℃ freezing condition was significantly reduced compared to -20℃ freezing condition in every sample except Japanese Spanish mackerel. There were less changes in color values of beef tenderloin, Japanese Spanish mackerel, radish in -70℃ condition than conventional freezing. The hardness of beef tenderloin, tofu was decreased and that of radish, blueberry was increased at -70℃ freezing in comparison to -20℃ freezing. By applying -70℃ freezing on beef tenderloin, we also found more decreasing tendency of volatile basic nitrogen compared to -20℃ freezing. In conclusion, the quality of general frozen foods, such as beef tenderloin, Japanese Spanish mackerel and blueberry, could be improved. The possibility of developing frozen tofu is also discovered by the application of liquid immersion freezing. These results could be meaningful as baseline data for researching and development of high-quality frozen foods.

PURPOSES: The purpose of this study is to identify the key factors that influence a driver’s trust in road-freezing risk information METHODS : A survey was designed and conducted in the metropolitan area to analyze drivers’perception towards road-freezing risk information. A binary logistic regression model was developed based on the survey responses. The statistically significant model coefficients were determined and their odds ratios were calculated. Finally, they were considered as the key factors influencing drivers’trust in roadfreezing risk information. RESULTS : The developed binary logistic regression model was validated through statistical tests. The final results indicated that previous experiences of accidents on slippery roads, as well as route choice patterns are statistically significant, having 95% confidence levels. CONCLUSIONS : Previous experiences of accidents on slippery roads and road choice patterns are the factors that most significantly influence drivers’trust in road-freezing risk information. The findings of this study will be used as fundamental data in establishing tailored road-freezing risk mitigation procedures and services.

최근에 국내에는 다양한 원인에 의한 구조물의 안정성을 우려하는 사회적인 관심이 생겼으며 이에 따른 구조물의 보수·보강에 다양한 연구가 진행되고 있다. 우리나라는 4계절이 뚜렷하며 특히 겨울과 봄에는 일교차가 심하게 발생하는 특징이 있다. 이러한 날씨는 철근콘크리트 구조물에게 동결융해작용을 발생시켜 성능저하의 원인이 되며 구조물의 안정성을 위협할 수 있다. 현재 구조물 보수·보강 방법으로는 탄소섬유나 유리섬유로 FRP(Fiber Reinforced Polymer)를 활용하여 Plate나 Sheet 형태의 부착 보강하는 방법이 일반화 되어있다. 하지만 다소 고가이며 유리섬유는 인체에 유해하다는 연구결과가 있다. 때문에 본 연구에서는 친환경적이고 내열성이 우수한 현무암섬유(Basalt Fiber)를 활용하여 동결융해 작용에 의한 콘크리트의 성능저하를 조건으로 BFRP-콘크리트의 부착성능 및 파괴패턴을 비교 분석하였다. 동결융해시험에 따른 부착강도평가는 동일한 섬유와 수지가 함침된 BFRP를 활용하여 동결융해Cycle(0, 100, 200, 300)과 콘크리트 압축강도(24MPa, 30MPa)를 변수로 부착성능을 평가하였으 며 Case1(선 부착 후 동결융해)과 Case2(선 동결융해 후 부착)로 나누어 진행하였다. Case1과 2 모두 콘크리트 파괴의 형태를 보 였으며 Case2의 경우 Case1에 비하여 부착강도가 감소됨을 나타내었고 동결융해 Cycle이 진행될수록 콘크리트 계면의 성능은 저하되고 부착강도는 평균 약 25%정도 감소하는 것을 확인하였다.

The aim of this study was to investigate the effect of pretreatments on quality of frozen peach. Pretreatments including steam blanching, water blanching, high pressure, and osmotic dehydration were applied to two varieties (Daeokgye and Hwangdo). Pretreated peaches were frozen and thawed at -20oC and 5oC, respectively. Steam blanching and osmotic dehydration with ascorbic acid reduced ΔE values without change of pH and acidity. Osmotic dehydration with sucrose decreased drip loss and increased brix. Freezing/thawing resulted in an increase of maximum force, while maximum force decreased with increasing time of steam and water blanching. Furthermore, osmotic dehydration with calcium chloride increased maximum force. High pressure decreased maximum force of Daeokgye and increased that of Hwangdo compared with non-treatment. Total polyphenol content, DPPH radical scavenging activity, and ABTS radical scavenging activity were improved by pretreatment including steam blanching, blanching, and osmotic dehydration with ascorbic acid.

The main purposes of this study were to identify the factors affecting the supercooling property and to improve the possibility of supercooling storage of fruits and vegetables. Freezing point and nucleation temperature, moisture content, hardness, sugar content, and pH of nineteen fruits and vegetables were measured and Pearson correlation analysis was performed. Freezing point showed a statistically significant correlation with moisture content and sugar content (p<0.01), while ice nucleation temperature showed a correlation (p<0.05) only for sugar content. In particular, the water content and sugar content did not show any correlation with the freezing supercooling difference (FSD). From the correlation analysis between FSD, aerobic bacteria, lactic acid bacteria, yeast, and mold, FSD showed a correlation (p<0.01) with aerobic bacteria. The experiments of the saline solutions inoculated with aerobic bacteria at different concentrations showed FSDs of about 2 for saline inoculated with 9.4 log CFU/mL and about 6 for saline inoculated lower than 5 log CFU/mL. Therefore, the aerobic bacteria concentration was determined to be a key factor affecting the supercooling storage of fruits and vegetables.

본 연구는 냉동전처리 기술이 분리막 기반의 슬러지 탈수공정에 미치는 영향에 대해 연구하였다. 냉동법은 응집법이나 원심분리법과는 달리, 슬러지 Cell lysis를 통해 Cell의 체액을 유출시킬 수 있음을 확인하였다. Cell lysis 이후 슬러지를 정밀여과법으로 분리하여 슬러지 탈수효율을 증가시키고자 하였으나, 냉동전처리로 인해 슬러지의 모세관 흡입시간과 여과소요시간이 약 13배 증가함으로써 슬러지 탈수효율이 감소하였다. 이는 슬러지로부터 방출된 유기물에 의한 슬러지 케이크 형성과 작은 슬러지 입자에 의한 세공차단현상에 의한 것이다. 적절한 공극 크기 및 슬러지 흡착에 내성을 갖는 분리막의 개발이 이루어 진다면, 냉동전처리와 분리막 공정의 혼성화기술은 슬러지 탈수에 적용 가능할 수 있을 것으로 보인다.

The present study was undertaken to evaluate the effect of trisaccharides supplementation in glycerol-free tris (GFT) for the cryopreservation of dog spermatozoa. In the first experiment (E1), dog spermatozoa were resuspended with 50, 75, 100 or 125 mM of raffinose, melezitose or maltotriose and cooled at 4 ℃ for 10 min. To determine the effect of different cooling time, the spermatozoa resuspended with 100 mM of raffinose, melezitose or maltotriose were cooled during 10, 20, 30 or 40 min at 4 ℃ (second experiment; E2). The straws were then aligned horizontally for 10 min on the rack and then plunged into LN2. In the third experiment (E3), to determine the effect of different vapor freezing time, the spermatozoa resuspended with 100 mM raffinose were cooled at 4 ℃ for 20 min and frozen in LN2 for 5, 10, 15 or 20 min and then plunged into LN2. In the fourth experiment (E4), to compare different freezing methods [cooling plus vapor freezing (CV), cooling plus step-down freezing (CS) and direct step-down freezing (SD)], the spermatozoa resuspended with 100 mM raffinose were cooled for 20 min and frozen in LN2 vapor for 5 min in case of CV method. In case of CS method, spermatozoa were cooled for 20 min at 4℃ and then frozen by the step-down freezing method. The straws were then aligned horizontally at 18, 15, 5, and 2 cm respectively from the surface of LN2 for 1, 1, 1.4, and 5 min, respectively in an L shaped straw holder and then plunged into LN2. For SD method, the straws were directly aligned horizontally at the same levels as CS from the surface of LN2 for 1, 1, 1.9, and 5 min, respectively and then plunged into LN2. After thawing at 37℃ for 25 sec, the spermatozoa were then incubated for 30 min in the freezing extender (E1) or in the 50 mM sucrose supplemented GFT (E2, E3, and E4) at 24℃. Following post-thaw incubation, sperm progressive motility and viability were assessed in E1, E2, E3, and E4. In addition, acrosome integrity, and gene expression related to apoptosis (BAX, BCL2, and Caspase10) and sperm motility (SMCP) were evaluated in E4. The results demonstrated that, in E1, using 75 mM trisaccharides resulted in significantly (p<0.05) higher sperm motility in all sugar groups. Using 100 mM melezitose significantly (p<0.05) improved the post-thaw viability than the 100 mM raffinose. The viability in 100 mM maltotriose was similar with 100 mM raffinose and melezitose group. In E2, the different cooling time has no significant effect on post-thaw sperm progressive motility in all the sugar types. In addition, the viability was variable among the different groups. In E3, liquid nitrogen vapor freezing for 5 min resulted in improved motility and viability. The sperm progressive motility was significantly (p<0.05) higher in CV and SD group compared to CS group and the sperm viability was significantly (p<0.05) higher in CV group compared to the other groups in E4. However, the acrosomal integrity of spermatozoa in the group CV was significantly (p<0.05) higher than the group CS and SD. In addition, the expression of SMCP gene was significantly (p<0.05) higher in the CV group than the CS group. In contrast, the expression of Caspase10 significantly (p<0.05) lower in the group CV and SD than the group CS. Furthermore, the ratio of gene expression of BAX and BCL2 was significantly (p<0.05) lower in the group CV than the group CS. Therefore, cryopreservation of dog spermatozoa in 100 mM of raffinose supplemented GFT cooled for 20 min and vapor freezing for 5 min provides better progressive sperm motility, viability, and acrosome integrity with higher expression of SMCP gene and lower expression of caspase10 and BAX/BCL2 ratio following post-thaw incubation in 50 mM sucrose supplemented GFT for 30 min at 24℃.

This study investigated the effect of various freezing and thawing conditions on the quality characteristics of radish. White radish was blanched and cooled down to corresponding freezing temperatures of -20℃, -40℃ and -60℃ and, thereafter, the radish was finally frozen in a -20℃ refrigerator for 24 h. The frozen radish was thawed at 4℃, 25℃ and with running water, radio-frequency thawing machine. As quality parameters, pH, drip loss, hardness, color, and sensory evaluation were evaluated. No effect of freezing and thawing conditions on color, pH, and total bacterial count was observed for the samples. Samples that underwent immersion freezing (-60℃) had lower drip loss and higher hardness than samples in other freezing conditions. Sensory evaluation showed that color, hardness, and overall acceptability increased with a decrease in freezing temperature. These results indicate that the temperature of freezing affects the quality of radish.

The present study was aimed to determine the effects of green tea extract (GTE) and beta-mercaptoethanol (β-ME) supplementation in boar sperm freezing extender on in vitro fertilization (IVF) and reactive oxygen species (ROS) and glutathione (GSH) levels of presumptive zygotes (PZs). Experimental groups were allocated into lactose egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/l in LEY) and β-ME (50 μM in LEY). In freezing, 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. The final sperm concentration was 1 × 108/ml. Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. For IVF, oocytes were matured in NCSU-23 medium and co-cultured with spermatozoa following thawing at 37°C for 25 sec. At 12 h following IVF, IVF parameters (sperm penetration and monospermy) were evaluated. In addition, GSH and ROS levels of PZs were determined by Cell Tracker Blue CMF2HC and DCHFDA, respectively. IVF parameters did not show any significant difference among the experimental groups. GSH and ROS levels of PZs were not significantly different between groups. In conclusion, antioxidant supplementation in boar sperm freezing could not influence IVF parameters, ROS and GSH levels of PZs.

In this experiment, we determined the effect of curcumin supplementation in freezing buffer for miniature pig sperm cryopreservation. Each ejaculate was diluted with modified Modena B extender and mixed with lactose-egg yolk (LEY extender, 80% v/v lactose solution [310 mM], 20% v/v egg yolk, and100 μg/mL kanamycin sulfate) and LEY-glycerol Orvus ES Paste (LEYGO, 89.5% v/v LEY, 5% v/v glycerol, 1.5% v/v Orvus ES Paste), 100 mM trehalose supplemented with 0, 10, 50, 100, and 500 μM of curcumin from turmeric, respectively. Following equilibration, the 0.5 mL French straws were frozen and plunged into LN2 tank for 7 days at least. Sperm parameter and oxidative byproducts were determined by the computer assisted sperm motility analysis (CASA) and fluorescence-activated cell sorting (FACS) as compared with each groups.Supplementation of curcumin had no effect on sperm motility, progressive motility and curvilinear velocity. However, average-path velocity and straight-line velocity were significantly higher in 10 μM curcumin group (100.9±8.8 μm/s, 61.7±2.9 μm/s, respectively) than control group (77.8±3.9 μm/s, 46.4±3.0 μm/s, respectively) (p < 0.05). In addition, the level of the O2 radical and H2O2 were comparatively decreased in curcumin groups by evaluation of ethidium and DCF fluorescence. According to the results, curcumin can improve sperm kinetic variables and alleviate ROS induced cryoinjury to pig sperm.

The present study was conducted to investigate the effect of different heights from liquid nitrogen (LN2) vapor on sperm motility and morphology after frozen-thawing. Two ejaculates were collected from 2 fertile Hanwoo bulls (A and B) by using artificial vagina at Hanwoo Research Institute. After collection, ejaculates were transferred to laboratory immediately and diluted with semen extender (Optixcell, France). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and cooled at 4°C for 4 h and loaded to 0.5 ml straws. The straws were divided into 2 groups. Straws were placed in 3 or 9 cm of LN2 vapor for 14 min and then plunged into LN2 tank and cryopreserved until evaluation. 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) after frozen-thawed. In bull A, 3cm group showed higher percentages of total motility, VSL with 25μm and VAP compared those with 9cm group (98.0 vs. 93.4%, 62.4 vs. 54.0% and 98.6 vs. 93.2%, 3 vs. 9 cm, irrespectively; p<0.001). In bull B, frozen-thawed sperm of 3cm group showed higher percentages of VSL with 25μm, VCL, VSL, VAP and BCF compared with those of 9cm group (43.5 vs. 26.0%, 123.8 vs. 111.6 μm, 62.9 vs. 57.3 μm and 81.5 vs. 72.5 μm; 3 vs. 9 cm, irrespectively; p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). In bull A, frozen-thawed sperm of 3 and 9cm groups showed no significant difference in LIA, LDA, DIA and DDA. In bull B, 3 cm group showed higher LIA and lower DIA compared with those of 9 cm group (73.2 vs. 23.7% and 23.7 vs. 32.2%, 3 vs. 9 cm, irrespectively; p<0.001). We suspected that 3 cm vapor on LN2 vapor might be affected positively spermatozoa viability and acrosomal integrity compared with 9 cm group. In conclusion, semen freezing procedure in the present study will improve sperm quality after frozen thawing.