• Animal Bioscience
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• v.35 no.9
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• pp.1351-1359
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• 2022

Objective: The objective of this study was to analyse the differentially abundant proteins caused by freeze-thawing of ram sperm and explore candidate proteins of interest for their ability to improve ram sperm cryopreservation outcomes in vitro. Methods: Sperm were from three mature Dorper. Fresh and frozen sperm proteins were extracted, and the differentially abundant proteins were analysed by mass spectrometry. Among these proteins, lactoferrin (LTF) was selected to be added before cryopreservation. Next, sperm samples were diluted in Tris extender, with the addition of 0, 10, 100, 500, and 1,000 ㎍/mL of LTF. After thawing, sperm quality was evaluated by motility, plasma membrane integrity, mitochondrial activity and reactive oxygen species (ROS). Results: Cryopreservation significantly altered the abundance of 40 proteins; the abundance of 16 proteins was increased, while that of 24 proteins was decreased. Next, LTF was added to Tris extender applied to ram sperm. The results showed that sperm motility and plasma membrane integrity were significantly improved (p<0.05) by supplementation with 10 ㎍/mL LTF compared to those in the control group. There was no significant difference in mitochondrial activity between the 0 ㎍/mL group and other groups (p>0.05). Supplementation of the cryoprotective extender with 10 ㎍/mL LTF led to decreased ROS levels compared with those in the control and other groups (p<0.05). Conclusion: The LTF is an important protein during cryopreservation, and the addition of 10 ㎍/mL LTF to a cryoprotective extender can significantly improve the function of frozen ram sperm.

• The Korean Journal of Blood Transfusion
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• v.23 no.2
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• pp.145-151
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• 2012

배경: 채혈 후 제제 경과시간에 따른 동결혈장의 혈액응고인자의 활성도와 이에 영향을 미치는 요인을 분석하여, 혈액응고인자제제의 원료혈장의 사용범위를 확장 가능한지 확인하고자 동결혈장의 채혈시간과 응고인자 특성을 측정하였다. 방법: ALT 부적격 혈장을 채혈 후 동결시간의 특성에 따라 4단계로 구분하였고, 6종류의 혈액 응고인자 활성도와 혈액형을 검사하였고, SAS 9.2 프로그램을 사용하여 통계처리 하였다. 결과: 혈액제제간 FVIII 활성도를 분석한 결과 PL-A>FFP>FP(8-24)${\approx}$FP(24-72) 순으로 유의하게 낮아졌고 혈액형에 따라서는 AB형이 제일 높고, O형이 제일 낮았다. 대한적십자사의 원료혈장에 대한 FVIII 활성 품질기준을 적용할 경우 PL-A, FFP와 FP24는 각각 85.0%와 82.5%로 적합하였다. 캐나다 퀘백 주처럼 FP24의 FVIII 활성이 0.52 IU/mL 이상을 적용할 경우 PL-A, FFP와 FP24는 각각 95.0%, 96.3%, 82.6%로 적합하였다. 또한 FP(8-24)의 A형과 AB형, FP(24-72)의 경우 AB형이 각각 82.1%, 83.3%, 100%로 적합하였다. 결론: 혈액응고인자제제용 원료혈장의 범위는 외국의 기준에 비추어 채혈 후 24시간 내에 동결된 혈장(FP24)으로 확대 사용이 가능하다. 이를 위해서는 채혈 후 동결시간과 혈액응고인자에 대한 품질기준을 유럽약전 또는 WHO 가이드라인과 비교하여 완화하는 것이 필요하다.

• The Korean Journal of Nuclear Medicine Technology
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• v.25 no.1
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• pp.29-33
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• 2021

Purpose Plasma renin activity (PRA) test is important for the diagnosis of primary aldosteronism. PRA is an easily deformed substance in vitro and affected by temperature changes. Laboratory of ASAN medical center has consistently found that there was a difference between the initial and re-experimental results. We compared and analyzed the differences in PRA test results according to the sample storage status. Materials and Methods The measurement of PRA was performed by using the radioimmunoassay. From August to September 2020, 43 PRA re-test samples were tested with different sample storage condition. The first group was re-examined by freezing the plasma-separated samples at -18℃, and the second group was re-examined with refrigerated EDTA sample. Also, additional tests were conducted on 13 PRA samples to verify the effect on thawing temperature differences in plasma-separated samples. The same samples were divided into two parts and stored frozen at -18℃, respectively, and thawing samples in room temperature and those in refrigerator were were conducted. Each result was compared and analyzed based on the initial experimental results. Results The results of re-examination after frozen storing plasma separation samples showed a lower correlation than the results of re-examination with EDTA plasma samples in refrigerator. When calculating the percentage based on the initial test results, the average percentage of each was 404.9% and 133.8%. The correlation coefficient was also R=0.8501 and R=0.9966, respectively, showing a higher correlation between plasma in the refrigerated sample EDTA tube. In comparison experiments with differences in thawing temperature, average percentage of the results of initial test and room temperature thawing was 94.3% and the average percentage of the results of refrigerated thawing was 88.0%. After again freezing the sample, the average percentage of the second room temperature thawing result is 107.5%, and the second refrigerated thawing group is 112.7%. Both groups showed an increase from first thawing. Conclusion A comparative analysis of retesting according to differences in sample storage methods in PRA tests showed a higher correlation between the results of retesting of the refrigerated EDTA plasma. And repeated freezing and melting of plasma separation samples, regardless of temperature during defrosting, has been shown to affect results. Therefore, retest of PRA should re-collect plasma from original EDTA plasma to increase reproducibility.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.5
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• pp.646-651
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• 2016

Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL) has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05) than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05). In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.2
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• pp.164-167
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• 2004

This study was carried out to investigate in vitro fertilization and development of in vitro matured pig oocytes inseminated with the Duroc boar sperm by different sperm washing media after thawing of the 5 ml frozen straws. Immature follicular oocytes (30-40) were transferred into each well of a Nunc 4-well multidish containing $500{\mu}l$ mTCM199 maturation medium. The sperm rich portion of ejaculates was collected into a 250 ml insulated vacuum bottle and gradually cooled 22 to $24^{\circ}C$ over a 2 h period. Semen was centrifuged at 800 g for 10 min and the seminal plasma discarded. Sperm were esuspended in a lactose-egg yolk and N-acetyl-Dglucosamine (LEN) diluent to contain $1{\times}10^{9}$ sperm/ml and cooled to $5^{\circ}C$ over a 2 h period. Immediately before freezing, semen was rediluted with an equal volume of LEN+4% glycerol and packed into 5 ml straws. After thawing of the 5 ml straw, the 5 ml semen was diluted with 20 ml Beltsville thawing solution (BTS) at room temperature. Oocytes were inseminated with untreated (unwashed and nonpreincubated) or treated sperm (washed two times in BTS, mTLP-PVA and mTBM media, respectively and nonpreincubated) with $2{\times}10^{7}$ sperm concentration. Oocytes were coincubated for 6 h in $500{\mu}l$ mTBM fertilization. At 6 h after IVF, oocytes were transferred into $500{\mu}l$ NCSU-23 culture medium for further culture of 6 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF and developmental ability of oocytes at 48 h after IVF were evaluated. Sperm penetration rate, male pronuclear formation and rate of cleaved embryos were higher in the BTS, mTLP-PVA and mTBM treatments than the unwashed treatment (p<0.05). The rate of blastocysts from the cleaved oocytes (2-4 cell stage) were higher in the mTLP-PVA treatment than in the unwashed, BTS and mTBM treatments. In conclusion, we recommend the washing of frozen-thawed sperm with mTLP-PVA medium before in vitro fertilization of oocytes in mTBM medium.

• Journal of Chest Surgery
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• v.29 no.3
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• pp.303-310
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• 1996

From December of 1994 to April of 1995, we, SHUH Department of Pediatric Thoracic and Cardiovascular Surgery, studied effects of aprotinin. 95 patients were randomly divided into two groups : group I (n=47) with aprotinin and group ll (n=48) without aprotinin. Aprotinin was given as one shot injection to cardiopulmonary bypass perfusion solution with dose of 50,000 KIUikg. Laboratory data such as hemoglobin, hematocrit, BUH, creatinine, fibrinogen, electrolyte concentration, aPTT, PT, and AT R was checked preoperatively, 5 minutes after anesthesia, 5 minutes and 35 minutes after CPB circulation, and 5 minutes, 3 hours, and 24 hours after reperfusion. Also, chest-tube drainage, transfused amount of RBC, platelet concentrate, and fresh frozen plasma within first 24 hours postoperatively were checked and analyzed after transition nn body weight demension. Only RBC transfused postoperatively had statistical significance with P value of less than 0.001. Others had no difference statistical wise. Postoperative side effects of aprotinin was not detected weeks after the surgery and there was no reoperated patient due to postoperative bleeding.

• Journal of Chest Surgery
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• v.29 no.2
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• pp.185-190
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• 1996

The effect of low-dose aprotinin on hemostasis in patients undergoing cardiopulmonary bypass (CPB) for repeat valve replacement and coronary artery bypass operations were investigated. Thirty patients undergoing elective CPB from February 1993 through February 1995 at Catholic Medical Center were studied. the patients were randomly divided into two groups(15 patients per group) : group 1, receive 1, 000, 000 KIU/kg aprotinin in the CPB priming volume and 20, 000 KIU/kg aprotinin intravenously each hour during CPB ; group 2, without aprotinin administration served as the controls. The result showed that the early postoperative (during the first 24 hours) and mean postoperative total blood loss of the aprotinin group were significantly reduced than the control group (317.2 $\pm$ 89.6ml in the aprotinin group versus 821.3 $\pm$ 441.2rnl in the control group, p<0.01 ; 767.2 $\pm$ 214.1 ml in the aprotinin group versus 1562.5 $\pm$ 735.2 rnl in the control gorup, p<0.01). Total use of packed red ells and fresh frozen plasma was higher in control group(1.22 $\pm$ 0.3 units versus 4.21 $\pm$ 1.7units of packed red cells, p<0.01 : and 2.37 $\pm$ 0.4units versus 6.72 $\pm$ 0.88uni1s of fresh prozen plasma, p<0.05). We conclude the low-dose aprotinin was positive influence on postoperative blood loss in undergoing highly bleeding potency cardiac operation.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.270-277
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• 2018

Objective: The present experiment has tested the effect of dietary alfalfa flavonoids (AAF) supplementation on the productive performances, carcass characteristics, meat quality and lipid oxidation of growing rabbits. Methods: One hundred and sixty crossbred rabbits (42 days old) were divided into four groups of forty animals each and were fed either a control diet (AAF0) or an AAF0 diet supplemented with 400, 800, or 1,200 mg of AAF/kg per diet (AAF4, AAF8, and AAF12, respectively) from weaning to slaughtering (102 days old). Performance data were recorded over a period of 60 days. At the end of the trial, 12 rabbits were slaughtered per group, and the carcass characteristics were recorded. Moreover, the plasma, liver and dorsal muscles were sampled from 12 rabbits/group, and were analyzed for lipid oxidation. Results: No significant differences were recorded for the performance, carcass characteristics and meat quality traits except for lightness parameter that was lower in the control group. Dietary AAF supplementation significantly (p<0.01) affected the malondialdehyde (MDA) levels of the frozen meat in a dose-related manner, with the lowest value (0.24 mg MDA/kg fresh meat) recorded in the AAF12 group samples. Conclusion: These findings indicated that the dietary inclusion of AAF in rabbit diets improved muscle oxidation stability with no adverse effects on the growth performance of the animals even if a slight impact on meat lightness color parameter was recorded.

• Korean Journal of Animal Reproduction
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• v.25 no.3
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• pp.269-275
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• 2001

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, RSP-S and RSP-T semen and fractional semen of small size dogs, and the effect of temperature and preservation time and cryoproservation on motility of whole and RSP-S and RSP-T semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. The volume per ejaculate semen, sperm of concentration and motility and abnormal sperm rate of 1st fractional semen were 0.65$\pm$0.09 $m\ell$, 4.52$\pm$0.35$\times$10$^{6}$ cells/$m\ell$, 15.64$\pm$3.85% and 5.50$\pm$0.62%. Also, 2nd fractional semen were 1.25$\pm$0.20$m\ell$, 3.35$\pm$0.48$\times$10$^{6}$ cells/$m\ell$, 96.25$\pm$4.65% and 4.24$\pm$0.46%. And 3rd fractional semen were 1.45$\pm$0.21$m\ell$, 3.55$\pm$0.52$\times$10$^{6}$ cells/$m\ell$, 92.82$\pm$4.24% and 4.66 $\pm$0.58%, respectively. 2. The sperm of concentration and motility and abnormal sperm rates of whole, RSP-S and RSP-T semen were 5.45$\pm$0.82$\times$10$^{6}$ cells/$m\ell$, 95.55 $\pm$4.65%, 4.58$\pm$0.45% and 4.82$\pm$0.36$\times$10$^{6}$ cells/$m\ell$, 90.10$\pm$3.42%, 6.48$\pm$0.68% and 4.55$\pm$0.45$\times$10$^{6}$ cells/$m\ell$, 93.25$\pm$3.85%, 4.82$\pm$0.58%, respectively. 3. The motility of whole, RSP-S and RSP-T semen were higher at 4$^{\circ}C$ than at 38$^{\circ}C$. When preservation temperature was at 4$^{\circ}C$, survival rates of RSP-S and RSP-T sperm were 97.54~6.25% at 1~72 hrs, 97.40~5.62% at 1~100 hrs, respectively. 4. The survival rates of slow and rapid frozen 2nd fraction, RSP-S and RSP-T semen were 67.3$\pm$4.45%, 88.8$\pm$4.46% and 46.4$\pm$3.84%, 74.4$\pm$4.20%, respectively. Survival rates was significantly higher in frozen RSP-S and RSP-T semen than that in control group(8.5$\pm$2.12%).

• The Korean Journal of Blood Transfusion
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• v.29 no.3
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• pp.253-261
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• 2018

Background: A massive blood transfusion (MT) requires significant efforts by the Blood Bank. This study examined blood product use in MT and emergency O Rh Positive red cells (O RBCs) available directly for emergency patients from the Trauma Center in Ajou University Hospital. Methods: MT was defined as a transfusion of 10 or more RBCs within 24 hours. The extracted data for the total RBCs, fresh frozen plasma (FFP), platelets (PLTs, single donor platelets (SDP) and random platelet concentrates (PC)) issued from Blood Bank between March 2016 and November 2017 from Hospital Information System were reviewed. SDP was considered equivalent to 6 units of PC. Results: A total of 345 MTs, and 6233/53268 (11.7%) RBCs, 4717/19376 (24.3%) FFP, and 4473/94166 (4.8%) PLTs were used in MT (P<0.001). For the RBC products in MT and non-MT transfusions, 28.0% and 34.1% were group A; 27.1% and 26.0% were group B; 37.3% and 29.7% were group O, and 7.5% and 10.2% were group AB (P<0.001). The ratios of RBC:FFP:PLT use were 1:0.76:0.72 in MT and 1:0.31:1.91 in non-MT (P<0.001). A total of 461 O RBCs were used in 36.2% (125/345) of MT cases and the number of O RBCs transfused per patient ranged from 1 to 18. Conclusion: RBCs with the O blood group are most used for MT. Ongoing education of clinicians to minimize the overuse of emergency O RBCs in MT is required. A procedure to have thawed plasma readily available in MT appears to be of importance because FFP was used frequently in MT.