본 연구에서는 염색⋅염료 산업의 폐수 처리 및 재활용 공정에 분리막을 적용하고자 실란 커플링제를 이용하여 상용화된 나노복합막을 표면 개질하였다. 실란 커플링제는 말단 관능기가 다른 octyltrimethoxysilane (OcTMS)와 (3-aminopropyl) trimethoxysilane (APTMS)을 사용하였으며, 표면 개질을 통해 염료 분리 및 내오염성을 향상시키고자 하였다. XPS, FE-SEM, EDX 분석을 통하여 막 표면의 화학 구조 변화 및 실란 적층을 확인하였고, AFM 분석을 통해 개질막의 표면 모폴로지를 확인하였다. Zeta potential을 통해 실란 개질 막이 상용막 대비 표면 전하가 중성으로 변하는 것을 확인하였다. 그 결과, OcTMS와 APTMS로 개질한 막의 내오염성은 NE70에 비해 약 2배 이상 향상되었다. 또한, 실란으로 표면 개질한 나노복합막은 음이온 염료(Orange II) 용액에서 약 90% 이상, 양이온 염료(Safranin-O) 용액에서 약 98% 이상의 염료 제거율을 나타내어 양이온 염료 용액 처리에 적합한 것을 확인하였다.

1. 본 연구는 한국 재래종 강낭콩 209자원의 phytochemical 및 항산화활성을 평가하였다. 2. 항산화활성은 DPPH, ABTS, FRAP, SOD를 분석하였으며 phytochemical은 kaempferol, myricetin, quercetin, naringenin 함량을 각각 분석하였다. 3. 항산화활성은 강낭콩 자원 간 다양한 분포를 보였으며 DPPH의 경우 62.3~643.9 (IC50), ABTS의 경우 0.28~1.49 mgAAE/g, FRAP의 경우 0.41~5.44 mgAAE/g, SOD의 경우 50.4 ~ 299.8 (IC50)로 나타났다. 4. Relative antioxidant capacity index (RACI)로 강낭콩 자원의 항산화활성을 비교한 결과 IT104587이 가장 높은 항산화활성을 보였으며 IT189598이 가장 낮은 항산화활성을 보였다. 5. 분석된 Phytochemical 중에서 한국 재래종 강낭콩에서는 Kaempferol이 가장 높은 함량을 나타냈다. 6. PCA 분석 결과 209자원은 3개의 그룹으로 나뉘었으며 이중 그룹 III에 속한 46자원의 강낭콩이 낮은 항산화활성 및 phytochemical 함량을 보였다. 7. 본 연구 결과는 한국 재래종 강낭콩의 항산화활성 및 phytochemical 정보를 제공하며 이 정보는 강낭콩 품종 개발을 위한 기초 정보로 사용될 수 있을 것이다.

Ganglioside GM3 is known as an inhibition factor of cell differentiation and proliferation via inhibition of epidermal growth factor receptor (EGFR) phosphorylation. Our previous study showed that the exogenous ganglioside GM3 reduced the meiotic maturation of porcine oocytes and induced apoptosis at 44 h of in vitro maturation (IVM). However, the role of ganglioside GM3 in the relationship between EGFR signaling and apoptosis during porcine oocyte maturation has not yet been studied. First, porcine cumulus-oocyte complexes (COCs) were cultured in the NCSU-23 medium with exogenous ganglioside GM3 according to maturation periods (non-treated, only IVM I: 0 - 22 h, only IVM II: 22 - 44 h and IVM I & II: 0 - 44 h). We confirmed that the proportion of germinal vesicle breakdown (GVBD) increased significantly in the IVM I treated group than in the control group. We also confirmed that the meiotic maturation until M II stage and polar body formation decreased significantly in the only IVM I treated group. Cumulus cell expansion and mRNA levels of the expansion-related factors (HAS2, TNFAIP6 and PTX3) decreased significantly in the IVM I treated group than in the control group. Protein levels of EGFR, p-EGFR, ERK1/2, and p-ERK1/2 decreased significantly in the GM3-treated groups, during the IVM I period. In addition, cellular apoptosis, determined using TUNEL assay, and protein levels of Cleaved caspase 3, were increased significantly in the GM3-treated COCs during the IVM I period. Based on these results, ganglioside GM3 exposure of porcine COCs during the IVM I period reduced meiotic maturation and cumulus cell expansion via inhibition of EGFR activity in pigs.

Here, we evaluated the mode of programmed cell death during porcine oocyte maturation by comparing the two major pathways associated with programmed cell death, apoptosis (type I), and autophagy (type II). We investigated the expression and localization of major genes involved in autophagy and apoptosis at mRNA and protein levels. Furthermore, the effect of hormonal stimulation on autophagy and apoptosis was analyzed. We found that the activity of autophagy-associated genes was increased in the cumulus-oocyte complexes (COCs) following follicle-stimulating hormone (FSH) treatment, while the addition of luteinizing hormone (LH) reversed this effect. The expression of proteins associated with autophagy was the highest in FSH-treated COCs. On the other hand, caspase-3 protein level was maximum in COCs cultured with LH. The treatment with rapamycin resulted in the effect similar to that observed with FSH treatment and increased autophagy activity. Thus, hormonal stimulation of pig oocytes resulted in distinct patterns of maturation. The high-quality oocytes majorly relied on the type II pathway (autophagy), while the type I pathway (apoptosis) was more prominent among poor-quality oocytes. Further investigation of this distinction may allow the development of techniques to produce high-quality oocytes in porcine in vitro fertilization.

게임의 상업적 성공에 힘입어 게임 원작의 애니메이션도 제작이 되었는데, 특히 1994년 <스트리트 파이터 II>로 시 작해 현재까지 꾸준히 게임을 원작으로 한 실사판, 2D Animation, 3D Animaion의 영상이 다양하게 제작되고 있다. 그러나 대중적, 상업적으로 성공한 게임을 원작으로 애니메이션이 제작된다고 항상 흥행에 성공하진 않는다. 그 이유는 원작 게임이 애니메이션으로 다시 제작 되었을 때, 원작 게임 스토리의 서사 구조, 그리고 게임과 애니메이 션의 설정에 대한 차이점이 두드러지기 때문이다. 그럼에도 불구하고 2016년 5월 19일에 개봉된, <앵그리버드> 게임을 원작으로 한 <앵그리버드 더 무비> 장편 애니메이션은 상업적으로도 큰 성공을 거두었는데, 본 논문에서는 크리스토퍼 보글러의 영웅의 여행 12단계가 <앵그리버드 더 무비>에 적용된 사례를 분석하고 다른 애니메이션과 비교하여 게임을 원작으로 한 애니메이션의 효과적인 서사 구조 발전방향에 관해 논의하고자 한다. 크리스토퍼 보글러는 헐리우드의 극작가로 신화 연구자인 조셉 캠밸의 <천의 얼굴을 가진 영웅>을 주 모티브로 신화의 공통된 서사 방식을 바탕으로 대중의 사랑을 받는 영화의 구조를 분석했다. 그가 정리한 서사방식은 영웅의 여행으로써 총 12 단계의 서사구조를 통해 이야기 속 주인공이 자아를 찾고 원하는 바를 이루어내는 과정을 시나리오 작법에 응용했다. 해외의 영웅, 모험 영화들 뿐만 아니라 우리가 잘 알고 있는 디즈니, 픽사, 지브리 등 대형 제작사에서 나온 애니메이션들도 이 크리스토퍼 보글러의 이야기 전개 방식을 많이 응용하고 있다.

본 연구의 목적은 스마트 헬스케어를 위해 접촉식 직물전극의 구조가 심장활동 신호 획득에 미치는 영향을 연구하는 것이다. 본 연구에서는 심장활동 신호 측정을 위하여 전극의 크기와 구성방식을 조작한 6종의 접촉식 직물전극을 컴퓨터 자수 방식으로 구현하였고, 이를 가슴밴드에 부착하여 응용형 리드 II(modified Lead II) 방식으로 심장활동 신호를 검출하였다. 건강한 신체의 남성 4명을 대상으로 서서 정지한 자세에서 각 직물전극을 사용하여 심장활동 신호를 검출하였으며, 모든 유형의 전극에 걸쳐 4회씩 반복측정 하였다. 심장활동 신호의 수집을 위해 BIOPAC ECG100 장비를 사용하여 1 ㎑로 샘플링하였으며, 검출된 원 신호를 대역통과 필터를 사용하여 필터링하였다. 직물전극의 구조에 따른 심장활동 신호 획득의 성능을 비교하기 위하여 신호의 파형과 크기를 파라미터로 하여 정성적 분석을 실시하였고, 각 전극을 통하여 획득된 심장활동 신호의 SPR(signal power ratio)을 산출함으로써 정량적 분석을 실시하였다. 산출된 SPR 값을 대상으로 하여 비모수 통계분석 방식의 차이검정과 사후검정을 실시함으로써 6개 전극의 구조에 따른 심장활동 신호 획득의 성능 차이를 구체적으로 분석하였다. 연구 결과 접촉식 직물전극의 구조에 따라 심장활동 신호의 품질에는 정성적, 정량적 측면에 걸쳐 모두 주요한 차이가 있는 것이 고찰되었다. 접촉식 직물전극의 구성 측면에 있어서는 입체전극이 평면전극에 비해 더 우수한 품질의 신호가 검출되는 것으로 나타났다. 한편 3가지 전극 크기에 따른 심장활동 신호 획득의 유의한 성능 차이는 발견되지 않았다. 이러한 결과는 심장활동 신호 획득을 위한 접촉식 직물전극 구조의 두 가지 요건 중 구성방식(평면/입체)이 웨어러블 헬스케어를 위한 심장활동 신호 획득의 성능에 주요한 영향을 미치는 것을 시사한다. 본 연구 결과를 기반으로 후속 연구에서는 직물전극이 일체형으로 통합된 의복형 플랫폼을 구현하고 성능 고도화 방안을 연구함으로써, 시공간의 제약 없이 고품질의 심장활동 모니터링이 가능한 스마트 의류 기술을 개발하고자 한다.

We report the properties of infrared photodetectors based on two kinds of quantum dots(QDs): i) 2.0 ML InAs QDs by the Stranski-Krastanov growth mode(SK QDs) and ii) sub-monolayer QDs by 4 × [0.3 ML/1 nm In0.15Ga0.85As] deposition(SML QDs). The QD infrared photodetector(QDIP) structure of n+-n−(QDs)-n+ is epitaxially grown on GaAs (100) wafers using molecular-beam epitaxy. Both the bottom and top contact GaAs layers are Si doped at 2 × 1018/cm3. The QD layers are grown with Si doping of 2 × 1017/cm3 and capped by an In0.15Ga0.85As layer at 495 oC. The photoluminescence peak(1.24 eV) of the SML QDIP is blue-shifted with respect to that (1.04 eV) of SK QDIPs, suggesting that the electron ground state of SML QDIP is higher than that of the SK QDIP. As a result, the photoresponse regime(~9-14 μm) of the SML QDIP is longer than that (~6-12 μm) of the SK QDIP. The dark current of the SML QDIP is two orders of magnitude smaller value than that of the SK QDIP because of the inserted Al0.08Ga0.92As layer.

The legacies of Tokyo Trial have been overlooked and questioned partly because prosecuting aggression was allegedly a violation of the principle of legality. This essay argues that the trial should not be overlooked for this reason because the legality debate at the trial provides insights into the interplay between the principle of legality and sources of international criminal law. Besides the majority judgment, some minority opinions could shed light on the nature of the Tokyo Charter by distinguishing between jurisdiction and applicable law and link the issue to the legality challenge. Although the Tokyo Charter was formally different from the Nuremberg Charter, both of them are substantive in nature so that the tribunals were allowed not to address the legality challenge. In addition, prosecuting aggression was arguably not a violation of the principle of legality because this principle, at that time, did not bind ex post facto legislation against international crimes committed during World War II.

본고는 고등학교 한문Ⅰ 교육과정 중 ‘이해와 감상’의 성취기준 중의 하나인 ‘한문 산문의 다양한 서술 방식을 통해 글의 내용을 이해하고 감상한다.’가 교과서에 어떻게 수용되었는지 그 양상을 살펴보고 문제점에 대하여 검토한 것이다. ‘한문 산문’의 ‘이해와 감상’을 포괄하는 성취기준은 ‘한문 산문의 다양한 서술 방식을 통해 글의 내용을 이해하고 감상한다.’이다. 교과서를 살펴본 결과 글의 감상에 한문 산문의 다양한 서술 방식을 이용한 경우는 그리 많지 않다. 많은 교과서가 한문산문에 사용된 서술방식만 제시하였고, 한문산문의 다양한 서술방식을 이용하여 산문을 감상한 경우에도 제대로 이루어지지 않고 있다. 이러한 사실은 고등학교 한문Ⅰ 수준에서 한문 산문의 감상은 제대로 이루어질 수 없음을 증명하는 것이다. 현실적으로 학생들이 한문을 이해함에 있어 글을 읽고 내용과 주제를 파악하는 것만도 너무 어렵게 느낀다. 이러한 학생들에게 산문의 이해와 감상은 할 수 있으 면 좋겠지만 무리한 요구라고 할 수 있다. 이는 2009 개정 한문과 교육과정이 반영된 교과서를 학습할 때에도 그 어려움이 그대로 드러난 점에서 확인할 수 있다. 따라서 한문의 본질인 읽고 내용 파악하는 데 중점을 두고 산문의 이해와 감상과 관련한 ‘한문 산문의 다양한 서술방식’은 한문Ⅰ 교육과정에서 삭제하거나 한문Ⅱ로 이동시키는 것이 바람직하다고 생각한다.

The ability of conventional semen analysis to predict male fertility is questionable. Since the prediction of male fertility is extremely of importance for the artificial insemination and profitable farm managements in animals, the development of highly sensitive biomarker of male fertility is a prime concern. Porcine Seminal Protein I (PSP-I) and Porcine Seminal Protein II (PSP-II) have been known that they are related with motility, and viability of spermatozoa. Thus, we investigated PSP-I and PSP-II level in boar spermatozoa to predict boar’s fertility. The expressions of PSP-I and PSP-II in spermatozoa from 21 individual boars with different fertility and litter size (litter size ranges from 10.3 – 14.2) were examined using qRT-PCR. Litter size was determined in 530 saws after artificial insemination (AI). In addition, sperm motility, motion kinematics, and capacitation status were measured using computer-assisted sperm analysis and Hoechst 33258/chlortetracycline fluorescence staining, respectively. PSP-I and PSP-II showed significantly negative correlation with litter size (r=0.578; P=0.006 and r=0.456; P=0.038, respectively). Furthermore, receiver-operating curves (ROC) was used to determine the accuracy for the prediction of boar fertility. Therefore we divided into 2 groups based on the median value of litter size. When selecting higher litter size group, PSP-I can predict litter size with overall accuracy 90.48% (sensitivity 88.89, specificity 91.67, negative predictive value 91.67, and positive predictive value 88.89) and PSP-II can predict with overall accuracy 81.82% (sensitivity 55.56, specificity 100.00, negative predictive value 76.47, and positive predictive value 100.00). Interestingly, PSP-I and PSP-II were found to increase 0.76 pups than average litter size (average 12.48) in tested boars. To best of our knowledge, this study is the first trial to investigate the correlation between PSP-I, PSP-II, and litter size. Therefore, we suggest that PSP-I and PSP-II could be considered as promising biomarkers for predicting male fertility and litter size outcome in field condition.

The lutropin/chorionicgonadotropin receptor (LHR) is a member of the rhodopsin-like subfamily of G protein-coupled receptors (GPCRs) that have been shown to mediate the internalization of its five (activation: three; inactivation: two) naturally occurring mutation. Gonadotropin receptors are members of the seven transmembrane (TM) receptor families. Several point mutations in TM II, III, V and VI have been identified in the luteinizing hormone receptor (LHR) gene, leading to constitutive activation and inactivation of the receptor. In eelLHR, we generated 3 types of constitutive activating mutations (M410T, L469R and D590Y) and 2 types of constitutive inactivating mutations (D383N and Y546F) to investigate how they work on hormone-receptor interaction. To assess the functional effects of 5 receptor mutations directly, wild-type (WT) and mutant receptors were transiently expressed in CHO-K1 cells. We evaluated the basal and cAMP stimulation by rec-LH hormone. The activity was shown to be a dose-dependent increase in cAMP production in LHR-WT expressing cells with an EC50 of 24.3 ng/ml and basal cAMP level of 2.6 nM. However, three activation mutants (D590Y, L469R and M410T) was most elevated the basal cAMP response at 12.8, 21.7 and 6.1 nM, respectively. In two inactivation mutants (D383N and Y546F) are very low in the basal cAMP activation. The EC50 was also considerably decreased to 42.3 ng/ml and 1181 ng/ml, respectively.

This study was conducted to evaluate the effects of insulin and epidermal growth factor (EGF) in a in vitro growth (IVG) medium on oocyte growth, in vitro maturation (IVM) and embryonic development of pig oocytes derived from small antral follicles (SAF) less than 3 mm in diameter. SAF oocytes were cultured for 2 days to induce IVG in alpha-minimal essential medium supplemented with 1 mM dbcAMP and 15% (v/v) fetal bovine serum. After IVG culture, oocyte maturation was induced by culturing IVG oocytes in IVM medium for 44 h. IVM oocytes that extruded the first polar body were selected and induced for parthenogenesis (PA) by applying electric stimulus. SAF oocytes cultured under the insulin treatment showed a significantly increased (P < 0.05) nuclear maturation (73.8%) compared to those cultured with insulin and EGF (59.8%). After PA, the proportions of blastocysts based on the number of metaphase II oocytes were significantly higher (P < 0.05) in oocytes that were cultured for IVG with insulin, EGF, and insulin + EGF (32.4%, 35.2%, and 34.8%, respectively) than in control (22.9%). IVG oocytes treated with insulin showed an increased oocyte diameter (116.3 μm) compared to those treated with insulin and EGF (114.0 μm) (P < 0.05). Intra-oocyte GSH content significantly increased (1.07 pixels/oocyte) by insulin treatment during IVG compared to that of oocytes treated with insulin + EGF (0.78 pixels/oocyte). These results demonstrate that IVG culture of SAF oocytes under insulin or/and EGF treatment supports oocyte maturation and improves embryonic development to the blastocyst stage after PA in pigs.

U0126 is a highly selective inhibitor of both MEK1 and MEK2, a type of MAPK/ERK kinase. This study was conducted to evaluate the effect of U0126 treatment during in vitro maturation (IVM) on nuclear maturation, intra-oocyte glutathione content, and embryonic development after parthenogenesis (PA). U0126 (5 μM) was supplemented to IVM medium during the first 0 (control), 2, and 4 h. The basic medium used for IVM was medium-199 supplemented with 10% (v/v) porcine follicular fluid (standard), 0.6 mM cysteine, 0.91 mM pyruvate, 75 μg/ml kanamycin, and 1 μg/ml insulin. Immature pig oocytes were matured for 44 h and then oocytes reached metaphase II stage were electrically activated to induce PA. The in vitro culture medium for embryonic development was porcine zygote medium-3 containing 0.3% (w/v) fatty acid-free BSA. When immature oocytes were treated with U0126 during the first 0, 2, 4 h of IVM culture, nuclear maturation was significantly (P < 0.05) increased by the U0126 treatment for 4 h (96.2 ± 1.3%) compared to standard IVM (90.6 ± 2.1%). Cleavage of PA embryos was significantly increased by 4 h- treatment (90.6 ± 2.2%) compared to standard medium (83.9 ± 1.8%). In addition, blastocyst formation of PA embryos was significantly (P < 0.05) increased by the treatment for 4 h (55.8 ± 5.7%) compared to 2 h (38.1 ± 6.1%). The glutathione contents in IVM oocytes were not altered by the U0126 treatments for 0, 2, and 4 h (1.28 ± 0.10, 1.16 ± 0.09, and 1.10 ± 0.09, respectively). Our results demonstrated that 5 μM U0126 treatment during the first 4 h of IVM showed positive effects on nuclear maturation, cleavage, and embryonic development in pigs.

Alpha-linolenic acid (ALA; n-3 18:3), a one of omega-3 fatty acid, is mainly contained in chloroplast of plant and ALA is an essential fatty acid, not synthesized in mammalian body, it must be supplied from foods. Polyspermy is especially high on in vitro fertilization (IVF) in pigs, which is a major obstacle to in vitro embryo production systems. In our previous study, when ALA was supplemented during in vitro maturation (IVM), the methaphase-II rate and gluthathione level was increased. The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) supplementation during IVM and subsequent of IVF in pigs. The cumulus-oocyte complexes (COCs) were submitted to IVM medium containing 0, 25, 50, and 100 μM ALA for 44 h. After 44 h of IVM, denuded oocytes were co-cultured with spermatozoa during 18 h. After 18 h of in vitro fertilization, oocyte were using aceto-orcein method, to evaluated penetration rate, monospermy (number of monospermy oocytes/total oocytes), and the IVF efficiency (number of monospermy/total penetrated oocytes). In results, 25 and 50 μM ALA groups were significantly increased on penetration rate compared with 100 μM ALA group (p<0.05). Similarly, monospermy rate were significantly increased 25 and 50 μM ALA groups than control group (p<0.05). IVF efficiency was no significant difference between control and ALA treatment groups. Our findings suggested that treatment of ALA supplementation during in vitro maturation (IVM) and subsequent of in vitro fertilization in pigs, ALA can increase IVF efficiency by effectively blocking polyspermy and increasing monospermy some mechanism in porcine oocytes. However, the study of mechanism by which ALA blocks polyspermy are needed, and this study suggests that ALA has a positive effect on in vitro production of porcine oocytes by decreasing polyspermy. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).

The objective of this study was to identify the proteins actively involved in the protection and repair of damaged cells, secreted by canine adipose derived mesenchymal stem cells (AT-MSCs) into the conditioned media. For this purpose, conditioned media (CM) was recovered from passage three stage canine AT-MSCs and skin fibroblasts cultured in serum free media after 24, 48 and 72 h. The extraction of exosomes was performed from 10-20 ml of CM using total exosome isolation kit. The isolated exosomes were then subjected to western analysis for the identification of annexin-I, annexin-II, histone H3 and dysferlin proteins. Results demonstrated the expression of proteins in the conditioned media isolated from canine AT-MSCs reflecting their potential in reducing the extent of damage at cellular levels. In conclusion, the conditioned media derived from canine AT-MSCs can be helpful in restoring the normal structure of cells both in vivo and in vitro conditions.

일반적으로 세포·조직 및 장기이식 성공 예측은 수여자와 공여자간의 백혈구항원 일치도이고, 불일치 시 심각한 거부 반응을 유발함으로 세포치료제로 사용할 때 우선적으로 백혈구 항원일치도가 고려된다. 그러나 중간엽줄기세포(Mesenchymal Stem Cells, MSCs)는 다른 체세포와 비교하여 상대적으로 낮은 MHC I 항원발현과, 극히 낮은 MHC II 항원을 가지고 있으므로 동종세포치료제로서 주목을 받고 있다. 따라서 본 연구에서는 개 모델에서 MSCs 의 동종세포치료제로서 효능을 예측하기 위해 선행연구로 백혈구 항원(Dog Leukocyte Antigen, DLA)형 및 가계도내 일치도와 유전적다형성(Polymorphism) 을 분석하였다. DLA 분석을 위해 한가계도의 비글(Covance Beagles) 4 두(모견 1 두, 자견 3 두)로 부터 전혈을 채취하고, 밀도구배를 이용하여 백혈구만을 분리 후 DNA들을 각각 추출하였다. DLA 분석은 ClassII 유전자(DLA-DQA, DLA-DQB, DLA-DRB)에서 엑손 2 영역(약 300bp)을 증폭하고 Direct Sequencing 을 통해 밝혀진 염기서열을 NCBI Blast 와 IPD(Immuno Polymorphism Database)를 기반으로 하여 Universal nomenclature 에 따라 유전자형을 판독 하였다. 그 결과 DLA-DQA(022:01/022:01)와 DLA-DQB(107:01/102:01)는 4마리 모두 유전자형이 동일하였으나, DLA-DQB 는 각각 046:01/022:02, 03701/022:02, 00201/022:02, 03701/022:02 로 차이를 보였다. 이 결과를 통해 모견과 자견이 공통적으로 가지는 일배체형(Haplotype)은 DLA–DQA*022:01, DLA-DQB*022:02, DLA-DRB*102:01 이었음을 확인할 수 있었다. 그리고 일부 유전자의 염기서열에서 99% 유사도를 보이는 후보군들이 4 개씩 검색되었는데 이는 단일염기다형성(SNP)에 기인한 유전적다형성(Polymorphism)이 매우 높다는 선행보고들과 유사한 결과를 보였다. 본 실험결과는 향후 DLA 의 일치군과 비 일치군의 개중간엽줄기세포와 말초혈액단핵구세포(PBMC)들의 공배양을 통해 동종세포치료제 연구에 사용될 예정이다. * 본 성과물은 농촌진흥청 반려동물 연구사업(세부과제명 : 반려견에서 DLA 일치하는 줄기세포의 체외 치료능 평가, 세부과제 번호 : PJ013957022018)의 지원에 의해 이루어짐.

Sperm cryopreservation is well known as a valuable method to preserve the genetic traits. Although many studies have established semen cryopreservation protocols, lack of studies were conducted to discover the differences of sperm proteome and functions between ejaculated and epididymal spermatozoa following to cryopreservation. Therefore, the objective of this study was to (i) evaluate the effect of cryopreservation on bull epididymal spermatozoa and (ii) discover the potential biomarkers, which have highly tolerance to freezing on bull epididymal spermatozoa. Our preliminary study demonstrated that spermatozoa from each bulls have different resistance on freezing during cryopreservation. We divided spermatozoa into two groups according to sperm motility following to cryopreservation; high freezing-tolerant spermatozoa (HFS) and low freezing-tolerant spermatozoa (LFS). Several sperm functional parameters, i.e. sperm motility/motion kinematics, speed parameters, viability, mitochondrial membrane potential, and capacitation status. Our results showed that all parameters except for motion kinematics and capacitation status had significant differences between HFS and LFS. Subsequently, two dimensional electrophoresis were conducted to compare the expression levels of sperm proteome between both groups. Three proteins {glutathione s-transferase mu 5 (GSTM5), voltage-dependent anion-selective channel protein 2 (VDAC2), and ATP stynthase subunit beta (ATP1B1)} were differentially expressed. Based on these results, we propose that epididymal spermatozoa from individual bull have different freezability upon cryopreservation and three differentially expressed proteins might be selected as a biomarker to predict high freezing-tolerant epididymal spermatozoa.

Although some factors, including season, age, type of estrus (natural estrus vs. induced estrus) and semen type (conventional vs. sexed), affect the conception rate following artificial insemination (AI) in dairy cattle, there is little information about the influence of ovarian characteristics, such as preovulatory follicle (PF) location at estrus, on fertility in dairy cattle. In most breeds of cattle the right ovary appears to function more actively than the left and about sixty percent of pregnancies in dairy cattle occur in the right horn of uterus (Reece and Tuner, 1938). Our study aimed to compare conception rates in dairy cattle between PFs that developed in the left ovary and those that developed in the right ovary at estrus. In this study, we examined the locational effect (left or right ovary) of the preovulatory follicle (PF) on fertility in dairy cattle. In total, 955 artificial inseminations (AI) were analyzed. At AI, PF locations were examined using rectal palpation, and dairy cattle were divided into two groups on their PF locations: (i) the PF located in the left ovary (L-PF); and (ii) the PF located in the right ovary (R-PF). Pregnancy was diagnosed by rectal palpation or ultrasonographic examination 60 days after AI. The conception rate was 38.1% in all dairy cattle. Conception rate was higher in the R-PF (40.8%) than in the L-PF (33.2%). In summary, PF development in the right ovary was associated with increased conception rates in dairy cattle.