The α-Gal epitope (Galα1,3Galα1,4GlcNAc-R) is responsible for hyperacute rejection (HAR) during transgenic pig-to-non-human primate xenotransplantation. There are genes related to the expression of α-Gal epitope such as α1,3Galactosyltransferase gene (GT-/-) and the isoglobotrihexosylceramide synthase (iGb3s-/-). This study was performed to investigate the expression of α-Gal epitope in the skin derived from GT-/- transgenic pig. The skin (7/1000 inches) was obtained by dermatome (Zimmer® Electric Dermatome) from one month old of wildtype (WT) and GT-/- piglets, respectively. The skins were fixed, dehydrated, cleaned, and embedded. To analyze the expression of α-Gal epitope, the paraffin section of WT and GT-/- were stained with BS-IB4 lectin and isoglobotrihexosylceramide synthase antibody. There was a strong BS-IB4 lectin signal in the skin of WT, but not detected in GT-/-. However, the iGb3s positive signals were stained in the skin of both WT and GT-/-. Taken together, it can be postulated that the knocked out of GT gene may not enough to inhibit the expression of α-Gal epitope. Further studies are needed to evaluate the functions of the double knock out of GT and iGb3s on the expression of α-Gal epitope.

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy, if it occurs. Progesterone primes the endometrium for possible implantation and inhibits uterine contraction until birth. 20-alpha hydroxysteroid dehydrogenase (20α-HSD; EC.1.1.1.149) enzyme belongs to the family of aldo-keto reductases. 20α-HSD predominantly converts progesterone into its biologically inactive form 20α-hydroxyprogesterone (20α-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In addition, the activity of 20α-HSD during the luteal phase known to be inhibited by prolactin. In this study, we focused on the analysis of transgenic mice expressing EGFP under control of monkey 20α-HSD promotor in mice testis. The protein expression and localization were detected by Western blotting and Immunohistochemical analysis, respectively. 20α-HSD protein was detected at molecular weight of 37-kDa by Western blotting analysis and EGFP was found at 27-kDa in the testis of TG mice. Also EGFP and 20a-HSD protein expression on 1, 2, 4, 6 and 8 weeks after birth were assessed. Both of them were increased the expression level time-dependently. 20α-HSD were strongly expressed in seminiferous tubule from 1 week after birth as seen in Immunohistochemical analysis. However, EGFP was strongly expressed in the seminiferous epithelial cells. Then, we determined the expression of EGFP mRNA in mice testis. Using primers specific for mouse EGFP, mRNA expression levels were analyzed by RT-PCR. The EGFP molecular weights is 400bp, qRT-PCR results using EGFP primer, The Cq value of the ratio decreased as the age increased. On this basis, mRNA were increased the expression level time-dependently. In conclusion, these observations suggest that the 20α-HSD in testis could be play a pivotal role in the spermatogenesis.

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy, if it occurs. Progesterone primes the endometrium for possible implantation and inhibits uterine contraction until birth. Aldo-keto reductases (AKRs) belong to a superfamily of NADPH-dependent reductases that act on a wide range of substrates, including simple carbohydrates, steroid hormones, and endogenous prostaglandins. 20-alpha hydroxysteroid dehydrogenase (20α-HSD; EC.1.1.1.149) enzyme belongs to the family of aldo-keto reductases. 20α-HSD predominantly converts progesterone into its biologically inactive form 20α-hydroxyprogesterone (20α -OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In addition, the activity of 20α-HSD during the luteal phase is known to be inhibited by prolactin. We have been reporting on the molecular characterizations of placental and ovarian 20α-HSD in the bovine, pig, deer and monkey. In this study, we focused on the 20α-HSD expression in testis(6, 9, 12, 18 and 21 days after birth) of miniature pig. The protein expression and localization were detected by Western blotting and Immunohistochemical analysis. 20α-HSD protein was detected at molecular weight of 37-kDa by Western blot analysis. Also the RNA expression were detected by Reverse Transcription-PCR and quantification PCR. Additionally, We are going to analysis the function and role of 20α-HSD in the pig testis.

PP2A-B55α, a regulatory subunit of PP2A plays an important roles in regulating cell proliferation and survival. However, the functions for PP2A-B55α in mouse early embryo development is not clear. The objective of present were to investigate the expression patterns and to explore its biological function during mouse early development. Thetranscripts of PP2A-B55α were detected at all developmental stages in mouse embryo and decreased during embryo development. Immunostaining revealed that PP2A-B55α was present in both the nucleus and cytoplasm in early cleavage stage embryos. In the late embryonic development, PP2A-B55α was predominantly located in the cytoplasm. Knockdown (KD) of PP2A-B55α using double strand RNA not affect the proportion of cleaved embryos, but resulted in significantly decreased development to blastocyst stage and reduced total cell number in blastocyst. KD PP2A-B55α is able to induce sustained DNA damage and reduced the transcripts of non-homologous end joining (NHEJ) or homologous recombination (HR) pathways relative genes in mouse early embryo. KD PP2A-B55αcaused apoptosis and increase the transcripts of pro-apoptotic genes in blastocyst. Furthermore, The KDPP2A-B55α showed significantly lower cell proliferating rates (from 5-Bromo-deoxyuridineassayresults) in blastocysts and to talareas of out growth potential was decreased. These observation provide novel in sight into PP2A-B55α expression patterns in mouse early embryos and down-regulation of PP2A-B55α negatively impacted blastocyst development, total cell number, DNA damage, apoptosis, and proliferation and post-hatchingevents.

α-solanine is toxic to human health by disturbing digestive and central nervous systems. However, little information has been focused on investigated with respect to α-solanine influence in mammal oocyte maturation and quality. In this study, we investigated the effects of α-solanine on oocyte maturation, quality and possible molecular mechanisms in a pig model. Porcine Cumulus-oocyte complexes (COCs) were treated with increasing concentration (0, 1, 10, 20, 50 μM) of α-solanine subjected to further in vitro maturation culture. The result showed that α-solanine significantly inhibited cumulus cells expansion and increased oocyte death rates when the concentration of α-solanine more than 10 μM. After cell cycle and cytoskeleton analysis, the results showed that α-solanine (10 μM) disturbed meiotic resumption, increased abnormal spindle formation and cortical granules (CGs) distribution rates when compared with the untreated group. α-solanine (10 μM) triggered autophagy by increasing the expression of autophagy-related genes (LC3, ATG7, LAMP2) and accumulation of LC3-specific puncta (an autophagy maker). TUNEL staining assay showed that α-solanine significantly increased apoptosis in porcine oocytes confirmed by up-regulated the levels of BAX and CAPS3 genes. Further study revealed that exposure α-solanine (10 μM) to porcine oocytes induced ROS generation, reduced mitochondrial membrane potential. In addition, our results suggested that α-solanine (10 μM) significantly increased the levels of H3K36me3 and H3K27me3 in porcine oocytes. Taken together, these data indicated that α-solanine toxic impaired oocyte maturation and quality by inhibited cumulus cells expansion, increased abnormal spindle and CGs distribution rates, triggered autophagy/apoptosis occur, accumulated ROS, decreased mitochondrial membrane potential, and changed epigenetic modifications.

The HNF4α (hepatocyte nuclear factor 4, alpha) is a hepatic transcription factor related to the lipid metabolism and regulation of insulin secretion in humans. The current study about commercial broiler reported that the A543G single nucleotide polymorphism (SNP) within HNF4α gene has an effect on fat deposition and wing yield in the chicken. This study was performed to investigate the association between the SNP within HNF4α gene and growth trait and to verify the applicability as a molecular marker for the improving the performance in Korean native chickens (KNCs). A total of 764 KNCs was collected from the livestock farm of Gyeongnam National University of Science and Technology in Korea and genotyped by PCR-RFLP. The body weight measured at birth, 2, 4, 6, 8, 10, 12, 14, 15, 16, 18, 20, 24, 28, 32, 36, and 40 weeks of age for the association analysis between chicken growth and the SNP genotype. Statistical analysis of the SNP with phenotypic traits was performed using SAS program. The KNC strain was classified by the genotypes of the A543G SNP. The frequencies of three genotypes were 0.47 (AA), 0.43 (AG) and 0.10 (GG), respectively. The SNP of HNF4α has highly significant association (p<0.001) with all-round growth in KNCs. These results suggest that the A543G SNP within HNF4α gene could be a genetic marker for the breeding in Korean native chickens.

An amide group was introduced to restrain the cohesion of silica nano-particles and copolymerized with polyamic acid. Amide block copolymers were prepared using silica and (3-mercaptopropyl) trimethoxysilane (MPTMS) with a siloxane group, using 2, 6-Lutidine as a catalyst. Amide block polymers and copolymers were synthesized via ATRP after brominating pyromellitic dianhydride (PMDA) and polyamic acid of methylene diphenyl diamine (MDA) using α-bromo isobutyryl bromide. Characteristic peaks of copolymer with amide and imide groups and patterns of amorphous polymers were studied using FT-IR and XRD analyses; an analysis of the surface characteristic groups was conducted via XPS. Changes in the thermal properties were examined through DSC and TGA; solubility for solvents was also studied.

알루미나 분말이 분산된 고분자용액을 비용매 유도 상전이법으로 방사 및 소결하여 알루미나 중공사막을 제조하 였다. 용매-비용매의 상호작용 속도에 따른 중공사막 기공 구조 형성을 확인하고, 특성을 분석하기 위해 dimethylsulfoxide (DMSO), dimethylacetamide (DMAc), triethylphosphite (TEP) 용매를 사용하여 방사액을 제조하였으며, 고분자 바인더로는 polyethersulfone (PESf), 첨가제로는 polyvinylpyrrolidone (PVP)를 사용하였다. 알루미나 중공사막의 기공 구조 변화를 확인 하기 위해 SEM으로 중공사막 단면을 분석하였다. DMSO, DMAc 용매를 사용할 경우 지상 구조(finger-like structure)와 망상 구조(sponge-like structure)가 복합된 기공 구조가 나타났으며, TEP 용매를 사용할 경우 전체적으로 망상 구조를 가졌다. 기공 구조에 따른 중공사막의 특성을 확인하기 위해 기체투과도, 기공도 및 기계적 강도를 측정하였다. 망상 구조를 갖는 중공사막 은 높은 기체 투과특성을 보였으며 지상 구조가 증가할수록 기체투과도가 감소하였다. 반대로 기계적 강도는 지상 구조가 발 달할수록 증가하였다.

본 연구에서는 입자크기가 다른 3가지 α-알루미나 분말로 부터 주입성형법과 소결법을 혼용하여 튜브형 α-알루미나 지지체 제작하였고 이때에 초기 α-알루 미나 분말의 입자크기와 소결 온도가 지지체의 기공구조와 기공구조가 투과 특 성에 미치는 영향을 고찰하였다. 제작 된 지지체는 수은함침법과 Archimedes 법을 통하여 기공경과 기공률을 측정하였다. 또한 30°C에서 He, N2, O2, CO2 기체에 대하여 투과 특성을 고찰하여, 각 지지체의 토튜오서티를 계산 하였으며, 지지체의 기공경 및 기공률이 지지체의 기체 투과 특성에 미치는 영향을 고찰 하였다.

본 연구에서는 α-glucosidase 저해활성이 우수한 천연 자원을 개발하기 위하여 개똥쑥으로부터 천연 물질을 분리·동정하고 분리된 물질에 대하여 α-glucosidase 저해활성을 측정하였다. 개똥쑥 잎과 줄 기의 chloroform층과 ethyl acetate층으로부터 각각 6개와 5개의 화합물을 분리하였다. 분리한 화합물 중 chloroform층으로부터 분리된 ArteCA(M.W. 316g/mol, C16H12O7)는 isorhamnetin, ArteCB(M.W. 360g/mol, C18H16O8)는 chrysosplenol D, ArteCC(M.W. 316 g/mol, C16H12O7)는 rhamnetin, ArteCD (M.W. 374g/mol, C18H16O8)는 chrysosplenetin, ArteCE(M.W. 284g/mol, C21H20O12)는 acacetin 그리 고 ArteCF(M.W. 270g/mol, C16H14O4)는 imperatorin으로 확인되었다. 한편 EtOAc층으로부터 분리된 ArteEAA(M.W. 178g/mol, C9H6O4)는 6,7-dihydroxy coumarin, ArteEAB (M.W. 192g/mol, C10H8O4) 는 scopoletin, ArteEAC (M.W. 448g/mol, C21H20O11)는 kaempferol-3-O-b-D-glucoside, ArteEAD (M.W. 464 g/mol, C21H20O12)는 quercetin-3-O-b-D-glucoside 및 ArteEAE(M.W. 318g/mol, C15H10O8)는 myricetin으로 확인되었다. 이 중에서 ArteEAE(myricetin)는 α-glucosidase에 대하여 97.3%의 높은 저해활성을 가지고 있었으므로, 혈당조절용 건강식품 또는 치료제 개발을 위한 물질로 활용될 수 있을 것으로 판단되었다.

Receptor activator of nuclear factor-κB ligand (RANKL) is an osteoblast/stromal cell-derived essential factor for osteoclastogenesis. During endochondral bone formation, hypertrophic chondrocytes calcify cartilage matrix that is subsequently resorbed by osteoclasts in order to be replaced by new bone. Hypoxia-induced upregulation of RANKL expression has been previously demonstrated in an in vitro system using osteoblasts; however, the involved mechanism remains unclear in chondrocytes. In the present study, we investigated whether hypoxia regulates RANKL expression in ATDC5 cells, a murine chondrogenic cell line, and hypoxiainducible factor-1α (HIF-1α) mediates hypoxia-induced RANKL expression by transactivating the RANKL promoter. The expression levels of RANKL mRNA and protein, as well as HIF-1α protein, were significantly increased in ATDC5 cells under hypoxic condition. Constitutively active HIF-1α alone significantly increased the levels of RANKL expression under normoxic conditions, whereas dominant negative HIF-1α reduced hypoxia-induced RANKL expression. HIF-1α increased RANKL promoter reporter activity in a HIF-1α binding element-dependent manner in ATDC5 cells. Hypoxia-induced RANKL levels were much higher in differentiated ATDC5 cells, as compared to proliferating ATDC5 cells. These results suggested that under hypoxic conditions, HIF-1α mediates induction of RANKL expression in chondrocytes; in addition, hypoxia plays a role in osteoclastogenesis during endochondral bone formation, at least in part, through the induction of RANKL expression in hypertrophic chondrocytes.

The viscosity, overrun, melting-down, moisture, crude fat, total sugar, and color of rice powder and puffed rice powder ice cream, following the addition of α-amylase, were investigated. For identical grain types, the gelatinization degree increased with puffing, and within the same treatment, the short grain was higher than the long grain. Viscosity dropped with increasing α-amylase at the same concentration and grain type, excluding 0.0%, the rice powder was higher than the puffed one, and for the same concentration and treatment, the short grain was higher. The overrun was highest at 0.2%, and for the same concentration and treatment, the short grain exhibited higher overrun. Higher melting-down was observed in puffed and lower viscosity ice cream mix. No significant difference was found in moisture with enzyme concentration. Regardless of puffing, the short tended to have a higher moisture. No difference was noticed in crude fat by concentration, grain type, or puffing. The total sugar was higher with increasing α-amylase; at the same concentration, puffed tended to be higher. The hunter “L” and “b” increased with α-amylase, while the “a” value dropped. At the same concentration and grain type, the “b” values decreased with puffing (p<0.05).

To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred α-1,3-galactosyltransferase knock-out (GalT —/—) pigs. In this study, the somatic cells and tissues from the GalT —/— pigs were characterized by an analysis of the expression of Galα-1,3-Gal (α-Gal) epitope. Briefly, ear fibroblast cell lines of 19 homozygous GalT —/— pigs were established and cryopreserved. The expression of α-Gal epitope in the cells was measured by fluorescence activated cell sorter (FACS) analysis using BS-I-B4 lectin. Also, the homozygous (GalT —/—) cells and tissues samples were immunostained with BS-I-B4 lectin for analysis of α-Gal epitope expression. The results showed that the expression of α-Gal epitope in GalT —/— cells (0.2 %) were significantly (p< 0.05) down-regulated to the range of cynomolgus monkey fibroblast (0.2 %) cells compared to heterozygous (GalT —/+) (9.3 %) and wild type (GalT +/+) (93.7 %) fibroblast cells. In the immunostaining results, while the expression of α-Gal epitope was detected a partly in GalT —/+ cells and mostly in GalT +/+ cells, it was almost not detected in the GalT —/— cells. Also, immunostaining results from various tissues of the GalT —/— pig showed that the expression of α-Gal epitope was not detectable, whereas various tissues from GalT +/+ pig showed a strong expression of α-Gal epitope. Our results demonstrated that α-Gal epitope expressions from GalT —/— pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

본 연구에서는 평균입경 0.2, 0.5, 1,7㎛ 크기의 α-알루미나 분말을 이용하여 다공성 α-알루미나 지지체의 기공구조를 조절하고자 하였다. 다공성 α-알루미나 지지체는 슬립캐스팅공법을 이용하여 제조한 후 소결하였으며, 이 때 소결 온도가 지지체의 기공특성에 미치는 영향에 대하여 고찰하였다. 제조된 다공성 α-알루미나 지지체는 수은기공분석기를 이용하여 기공크기 및 기공률 등을 분석하였으며, 단일기체투과장치를 이용하여 기체 투과도를 측정하였다. 그 결과 평균입경 0.2, 0.5, 1.7㎛ 크기의 α-알루미나 분말을 이용하여 제조된 지지체는 각각 80, 130, 200㎚의 기공경을 가졌으며, CO2 단일기체에 대해 각각 1300, 1700, 5000GPU를 나타냈다.

본 연구에서는 silicalite-1 제올라이트 분리막 합성 시에 종결정 코팅용액 pH 변화가 제올라이트 분리층 미세구조에 미치는 영항을 고찰하였다. 75 nm 크기로 합성된 종결정은 에탄올에 분산된 후 침지코팅법으로 지지체 표면에 코팅되었으며 분산용액의 pH는 2.2, 7.0, 9.3으로 조절되었다. pH가 7인 경우, 균일하고 두께가 3~4 μm인 silicalite-1 제올라이트 분리층이 형성되었고 분리층 결정입 크기는 100 nm로 미세하였다. 반면, pH가 2.2와 9.3인 경우, 분리층 두께가 얇고 불완전하였으며 분리층 결정입 크기도 약 1 μm로 조대하였다. pH 7에서 완전한 제올라이트 분리층이 형성된 것은 침지코팅 중에 지지체와 종 결정이 서로 다른 부호의 전하를 가져 정전기적 인력이 작용하여 균일하고 조밀하며 두껍고 다층의 종결정 코팅층이 형성되었 기 때문이었다. 반면에 pH가 2.2와 9.3인 경우, 침지코팅 중에 지지체와 종결정이 서로 같은 부호의 전하를 가져 정전기적 반 발력이 작용하기 때문에 불완전한 덮힘에 의하여 불완전한 분리층이 형성된다고 판단되었다. 결론적으로, 종결정 코팅용액의 pH가 silicalite-1 제올라이트 분리층의 두께, 결정립 크기 등 미세구조를 결정하는 중요한 인자임을 확인할 수 있었다.

변위전류 측정법을 L-α-dilauryl phosphatidylcholine(DLPC) 단분자 막의 연구에 적용하였 다. 변위전류는 물 표면에서 DLPC 단분자 층에서 압축과 확장에 의해 발생되었다. 맥스웰 변위전류 (MDC) 발생은 분자 당 점유면적 200 Å2 에서 40 Å2에 대하여 관찰하였다. 맥스웰 변위전류는 단분 자 층의 압축 사이클에 대해 조사하였으며, MDC의 최대 값은 압축 사이클의 표면 압력이 처음 상승하 기 바로 직전의 분자당 점유면적에서 나타나는 것을 알 수 있었다. LB막의 단분자층 표면 형태는 원자 힘 현미경(AFM)으로 측정하였다. 결과적으로, AFM 이미지에 나타난 LB막의 특성은 단분자 층의 배향 이 좋았으며 단분자 층의 두께는 약 5~10 nm였다.