The α-Gal epitope (Galα1,3Galα1,4GlcNAc-R) is responsible for hyperacute rejection (HAR) during transgenic pig-to-non-human primate xenotransplantation. To overcome HAR after xenografts, it is essential for the inactivation of α1,3Galactosyltransferase (GT) gene by the homozygotic knocked out of GT-/- and the isoglobotrihexosylceramide synthase (iGb3s-/-). This study was performed to investigate the generation and characterization of the α1,3GT-MCP/-MCP+iGb3-/- transgenic cells. Ear fibroblast cells from the GT-MCP/-MCP pig were cultured and used to positive control. For iGb3s knock out, the Cas9-GFP-iGb3s vector was transfected into the GT-MCP/-MCP cells. The Cas9-GFP-iGb3s transfected cells were sorted and sequenced for the selection of GT-MCP/-MCP+ iGb3s-/- cells. Among the three sorted cell lines, one transgenic cell line was homozygously deleted 3 bases and 10 bases in each chromosome, respectively. To characterize an expression of α-Gal epitope, a wild type and the transgenic cells were measured by FACS Aria using BS-IB4 lectin antibody. The expression of α-Gal epitope in GT-MCP/-MCP cells (<0.01 %) were significantly down-regulated to the range of wild type (99.4 %) fibroblast cells (p<0.05). To analyze the function of iGb3s, α -Gal epitope expressions were measured for the GT-MCP/-MCP, GT-MCP/-MCP+iGb3s-/+, and GT-MCP/-MCP+iGb3s-/-. The range was 95.8%, 94.2%, and 75.8%, respectively. Interestingly, there was a negative range (16.2%) of α-Gal epitope -/- section in GT-MCP/-MCP+iGb3s-/-, compared to 2.74% of GT-MCP/-MCP+iGb3s-/+ and 1.4% of WT, respectively. Our results demonstrated that iGb3s-/-combined with GT-/- had a function to inhibit α-Gal epitope expression in pig cells. Further studies are needed to evaluate the functions of double gene knock out to minimize a HAR response after xenotransplantation.

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.

RNA Sendai virus (SeV) vector system has no risk of being integrated into the host genome. Sendai virus (SeV) vectors expressing pluripotent factors have been used to produce integration-free induced pluripotent stem cells (iPSCs) with high efficiency from various cell types in human and mouse. In this study, we generated iPSCs from pig ear fibroblast cells using the SeV vector expressing 4 human factors (POU5F1, SOX2, C-MYC, and KLF4). Colonies were emerged at Day 14 of transduction and expressed the classical pluripotency markers (POU5F1, NANOG, and SOX2) and surface marker (SSEA1). Furthermore, they showed a domed shape and could passage over 40 times under 2i (CHIR99021 and PD0325901)-LIF and MEF feeder culture condition having in vitro differentiation ability into 3 germ layers. Next, we examined the ability of six feeder free culture conditions to maintain piPSCs in a pluripotent state. piPSCs were plated on Matrigel coated dishes in different media: 1. CM: control media (LIF culture media); 2. CM-F: CM+100 ng Fetuin-A; 3. CM-N: CM+100 ng Nanog-TAT; 4. CM-2i: CM+3 uM CHIR99021+1 uM PD0325901; 5. CM-2iN: CM-2i+100 ng Nanog-TAT; 6. CM-2iN+100 ng Fetuin-A. However, piPSC could not maintain the typical self-renewal morphology on feeder free conditions regardless of culture media tested here. Further, expression of pluripotency-related genes (Oct4, Nanog and Klf4) of piPSCs cultured on feeder free conditions could not be compared with that of iPSCs cultured on MEF feeder plate. Our results suggest that integration free pluripotent stem cell from pigs could be generated by SeV vector system and maintained their pluripotency under 2i-LIF and MEF feeder culture condition, but further optimization of culture conditions may be required.

The present study was conducted to investigate the effect of caffeine and theophilline on sperm motility during in vitro fertilization (IVF). Briefly, commercial boar semen was centrifuged and resuspended (5x107 sperms/ml) with fertilization medium (mTBM) supplemented with 2 mM caffeine (Caf), 5 mM theophylline (The), 2 mM caffeine + 5mM theophylline (Caf + The), and not supplemented control. The semen parameters of the four groups were analyzed by computer-assisted semen analysis (CASA, Medical Supply Co. Ltd) system at 6 h as time for IVF at 38.5 C under 5 % CO2 in air. The groups were examined 11 semen parameters such as total motility (TM), curvilinier velocity (VCL), straight-line velocity (VSL), average-path velocity (VAP), and hyperactivated (HYP), etc. A total motility of control group (41.3 %) at 6 h showed significantly (P<0.05) higher than those of other groups (Caf, 37.2; The, 35.2; Caf + The, 30.1 %). Results from many other sperm parameters indicated that the theophylline and caffeine negatively affected on sperm motility during IVF. These results suggested that the supplementation of caffeine and/or theophylline was not essential for IVF in pigs. To prove this suggestion, further studies are needed to analyze fertility and embryonic development after IVF with or without the supplementation of caffeine and/or theophylline.

An experiment was conducted to study the potential of thermal treated oak sawdust(steaming and steam explosion) as horticultural medium component in plug seedlings production of Chinese cabbage(Brassica campestris L.). This study involves the chemical, physical characterization and growth test of thermal treated oak sawdust(steaming and steam explosion) in order to evaluate their use as components of horticultural media. A commercial peat moss and oak sawdust were used as control. The total carbohydrate, C/N ratio, pH, phenolic compound, total porosity and water holding capacity were 45.1g/100g dry wt, 425.1, 4.4, 141.8mg/g wt, 82.5%, 47.1% in oak sawdust and 39.2g/100g dry wt, 300.3, 4.7, 131.7mg/g wt, 84.9% and 49.2% in steamed oak sawdust and 30.3g/100g dry wt, 247.8, 5.7, 40.8mg/g wt, 92.3% and 51.7% in steam exploded oak sawdust, respectively. The mixtures of the horticultural media were prepared using different substrate as peat moss, oak sawdust, steamed oak sawdust, steam exploded oak sawdust and perlite to grow Chinese cabbage in a greenhouse. The seed germination, stem height and leaf area were 68%, 2.2cm, 1.1cm2 in OSP(containing 90% oak sawdust and 10% perlite) and 69%, 2.5cm, 1.5cm2 in SMP(containing 90% steamed oak sawdust and 10% perlite) and 87%, 3.0cm, 2.2cm2 in SEP(containing 90% steam exploded oak sawdust and 10% perlite), respectively. The leaf area SEP(containing 90% steam exploded oak sawdust and 10% perlite) was higher than that of PP(containing 90% peat moss and 10% perlite). This research indicates that steam exploded oak sawdust may be utilized as a suitable replacement for peat moss in horticultural media component for Chinese cabbage.

담낭 절제술 후 발생하는 혈액 담즙증은 심각하고 드문 합병증이며 일반적으로 수술 4주 이내에 발생하는 것으로 보고되고 있다. 저자들은 흑색변과 복통을 주소로 내원한 30년 전 개복 담낭 절제술의 과거력이 있는 77세 여자 환자 를 혈액 담즙증으로 진단하고 색전술로 성공적으로 치료하여 이를 보고하는 바이다. 또한 본 증례를 토대로 담낭 절제술 후 오랜 기간이 경과하였더라도 수술 후 혈액 담즙증이 발생할 수 있다는 것을 염두에 두어야 할 것이다.

Worldwide there is concern about the continuing release of a broad range of environmental endocrine disrupting chemicals, including polychlorinated biphenyls, dioxins, phthalates, polybrominated diphenyl ethers (PBDEs), and other halogenated organochlorines persistent organic pollutants (POPs) into the environment. They are condemned for health adverse effects such as cancer, reproductive defects, neurobehavioral abnormalities, endocrine and immunological toxicity. These effects can be elicited via a number of mechanisms among others include disruption of endocrine system, oxidation stress and epigenetic. However, most of the mechanisms are not clear, thus several number of studies are ongoing trying to elucidate them in order to protect the public by reducing these adverse effects. In this review, we briefly limited review the process, the impacts, and the potential mechanisms of dioxin/dioxin like compound, particularly, their possible roles in adverse developmental and reproductive processes, diseases, and gene expression and associated molecular pathways in cells.

This study aims to examine the effects of taping of the ankle joint on the static and dynamic balance and gait ability of stroke patients. Twenty-six stroke patients receiving physical therapy at a hospital located in Gyeonggi-do were divided equally into a group that had taping in physical therapy and an ordinary physical therapy group. They exercised for 30 minutes each, 3 times per week for 8 weeks from June to August 2011. Romberg’s eye open and eye closed tests, limits of stability(LOS), forward and back test, timed up and go test(TUG) and 10-meter gait velocity test were performed to evaluate static balance, dynamic balance, and gait ability, respectively, prior to and 8 weeks after the intervention. Differences within each group in relation to the lapse of time were compared by a paired t-test. Differences between the two groups were compared by an independent t-test. Regarding comparison of differences within each group, all tests resulted in significant changes in both groups after the intervention (p<.05). Comparison of differences between the two groups showed that taping in the physical therapy group had significantly better test results than the ordinary physical therapy group in all measured items(p<.05). The after effects of ankle taping on stroke patients are more efficient and effective than ordinary physical therapy alone in improving balance and gait ability.

강 상류에 설치된 대형 댐에 의해 연속성을 차단함으로써 유수역에서 정수역으로 변화된 금강 상류에 위치한 용담호를 선정하여 서식하는 어류상 및 어류의 군집 특성을 알아보기 위하여 2009년 4월부터 11월까지 조사를 실시하였다. 채집된 어류는 강 상류의 자연성을 잘 유지하고 있는 원서식지(St. 1)에서 8과 15종이며 고유어종은 11종이었고, 댐호로 인해 수환경 변화에 영향을 받을 것으로 생각되는 용담댐 직하류 수역(St. 3)에서 10과 24종이며 한국 고유종은 11종이 출현하였다. 반면 댐호로 인해 유수역에서 정수역으로 수환경이 변화된 용담호 내(St. 2)에서 7과 20종이며 한국고유종은 6종이 출현하였다. 금강 상류수역에 건설된 용담댐으로 인해 댐호에는 기존에 금강 상류수역의 대표적인 서식어류인 칼납자루, 감돌고기, 쉬리, 꾸구리 등과 같은 유수역에 서식하는 어류는 사라졌으며, 붕어, 끄리, 치리, 피라미, 블루길과 같이 정수역에 서식하는 어류들이 급격히 증가하는 경향이었다. 충식성 어류의 상대적 풍부도는 St. 1에서 66.7%인데 반하여 St. 2에서는 40.0%로 차이를 보였고, St. 3에서 중간 형태로 54.2%이었다. 우리나라의 생물자원인 고유어종의 보존뿐만 아니라 수생태계의 건강성 유지를 위해서는 강 상류에 대형댐과 같이 대규모의 상류 서식지를 변화시킬 수 있는 인공 시설물의 축조는 최소화하는 것이 바람직 할 것이며, 불가피한 경우는 건설시 보전대책 수립이 필요하다.

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to locate Ski protein in the rat ovary during luteinization to predict the possible role of Ski. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally.

Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. The aim of the present study was to locate Ski protein in rat ovaries in order to predict the possible involvement of Ski in follicular development and atresia. First, expression of c-Ski mRNA in the ovaries of adult female rats was confirmed by RT-PCR. Then, ovaries obtained on the day of estrus were subjected to immunohistochemical analysis for Ski and proliferating cell nuclear antigen (PCNA) in combination with terminal deoxynucleotidyl transferase- mediated dUTP nick end-labeling (TUNEL). Ski was expressed in granulosa cells that were positive for TUNEL, but negative for PCNA, regardless of the shape and size of follicles. Expression of Ski in TUNEL-positive granulosa cells, but not in PCNA-positive granulosa cells, was also verified in immature hypophysectomized rats having a single generation of developing and atretic follicles by treatment with equine chorionic gonadotropin (eCG). These results indicate that Ski is profoundly expressed in the granulosa cells of atretic follicles, but not in growing follicles, and suggest that Ski plays a role in apoptosis of granulosa cells during follicular atresia.

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to locate Ski protein in the rat ovary during luteinization to predict the possible role of Ski. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally.

Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. The aim of the present study was to locate Ski protein in rat ovaries in order to predict the possible involvement of Ski in follicular development and atresia. First, expression of c-Ski mRNA in the ovaries of adult female rats was confirmed by RT-PCR. Then, ovaries obtained on the day of estrus were subjected to immunohistochemical analysis for Ski and proliferating cell nuclear antigen (PCNA) in combination with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). Ski was expressed in granulosa cells that were positive for TUNEL, but negative for PCNA, regardless of the shape and size of follicles. Expression of Ski in TUNEL-positive granulosa cells, but not in PCNA-positive granulosa cells, was also verified in immature hypophysectomized rats having a single generation of developing and atretic follicles by treatment with equine chorionic gonadotropin (eCG). These results indicate that Ski is profoundly expressed in the granulosa cells of atretic follicles, but not in growing follicles, and suggest that Ski plays a role in apoptosis of granulosa cells during follicular atresia.

Antioxidants partially ameliorated the detrimental effects of reactive oxygen species (ROS) on sperm characteristics during in vitro storage. The objective of the present study was to investigate the single or synergetic antioxidative effect of curcumin and Vit. E on the characteristics of fresh boar sperm during in vitro storage. The sperm viability in curcumin, Vit. E supplementation and curcumin+Vit. E+H2O2 groups remained over 85.0% in 3 hr incubation period, but in 6 hr incubation period, curcumin+Vit. E+H2O2 groups was sharply dropped than those of curcumin and Vit. E group. The membrane intergrity in all evaluated groups except for H2O2 group did not significantly difference in 3 hr incubation period. The viability in curcumin or Vit. E supplementation were significantly increased than in curcumin+H2O2 and Vit. E+H2O2 group in 6 hr incubation period. The percentage of mitochondrial activity and acrosome intergrity obtained similar trends within same incubation periods irrespective of treatment. The lipid peroxidation of spermatozoal plasma membrane ranged from 11.6∼17.5 nM/l×106 and 14.0∼ 19.0 nM/l×106 in 3 hr and 6 hr incubation periods. In conclusion, curcumin or Vit. E rpplementation alone or cooperatively improved sperm viability index (motility, membrane intergrity, viability and survival rates) and fertility index (mitochondria activity, acrosome intergrity and lipid peroxidation) of fresh boar sperm, indicating that curcumin and Vit. E have a antioxidative properties through its scavenging activity against hydrogen peroxide.

Ixeris dentata is a typical oriental herb. It is a widely distributed perennial in Korea, Japan and China, which belongs to the Compositae Family. The whole plant of I. dentata has been used for the treatment of pneumonia, contusion, tumor and hepatitis.

The early growth response protein 1 (Egr-1) is a widely reported zinc finger protein and a well known transcription factor encoded by the Egr-1 gene, which plays key roles in many aspects of vertebrate embryogenesis and in adult vertebrates. The Egr-1 expression is important in the formation of the gill vascular system in flounders, which develops during the post-hatching phase and is essential for survival during the juvenile period. However, the complete details of Egr-1 expression during embryo development in olive flounder are not available. We assessed the expression patterns of Egr-1 during the early development of olive flounders by using reverse transcription polymerase chain reaction (RT-PCR) analysis. Microscopic observations showed that gill filament formation corresponded with the Egr-1 expression. Thus, we showed that Egr-1 plays a vital role in angiogenesis in the gill filaments during embryogenesis. Further, Egr-1 expression was found to be strong at 5 days after hatching (DAH), in the development of the gill vascular system, and this strong expression level was maintained throughout all the development stages. Our findings have important implications with respect to the biological role of Egr-1 and evolution of the first respiratory blood vessels in the gills of olive flounder. Further studies are required to elucidate the Egr-1-mediated stress response and to decipher the functional role of Egr-1 in developmental stages.