Induced pluripotent stem cells (iPSCs), generated by the overexpression of transcription factors Oct4, Sox2, Klf4 and c‐Myc in somatic cells, are pluripotent. iPSCs reprogrammed from differentiated cells get through a epigenetic modification during reprogramming and finally have the similar epigenetic state to embryonic stem cells (ESCs). In this study, these epigenetic changes were observed in reprogramming of uni‐parental parthenogenetic somatic cells. Furthermore, we have shown that parthenogenetic pattern of imprinted genes were changed during pluripotential reprogramming. Parthenogenetic neural stem cells (pNSCs) containing only maternal alleles regain the biparental imprinting patterns after reprogramming. However, we have yet to define whether the changed imprinted genes are maintained or reverted to the parthenogenetic state when the reprogrammed cells are differentiated into specialized cell types. To address this question, we compared genome‐wide expression profiles of biparental female neural stem cells (fNSCs), parthenogenetic neural stem cells (pNSCs), and NSCs differentiated from parthenogenetic maternal iPSC (miPS‐NSCs). Furthermore, this study establishes the correlation between the alteration of genome methylation and activation of imprinting genes in the parthenogenetic cells and reports for the first time that the silenced PWS‐related imprinted genes are activated in miPS‐NSCs. Our data demonstrated that pluripotential reprogramming of parthenogenetic somatic cells were able to reset the parthenogenetic imprinting patterns; reprogrammed miPSCs showed erasure of maternal methylation imprints and acquisition of methylation in paternally imprinted genes. Furthermore, the changed imprinting patterns were maintained when the reprogrammed cells are differentiated into specialized cell type. * This work was supported by the Next‐Generation BioGreen 21 program (Grant PJ008- 009) funded by the Rural Development Administration, Republic of Korea.

This study examined the effects of tamoxifen at different concentrations and treatment periods on proliferation of a human cervical cancer cell line, HeLa. As in the previous studies, Estrogen did not have an effect on the cellular proliferation at low concentrations but significantly reduced proliferation at high concentration (5 ug/ml) based on MTT assay. Treatment with tamoxifen had similar effects. It did not have any significant effect on the HeLa cell proliferation at low concentrations, but significantly reduced proliferation at high concentration (10 uM). In addition, combined treatment with both estrogen and tamoxifen did not alter the inhibitory effects of either estrogen or tamoxifen on cellular proliferation. The inhibitory effects by tamoxifen on HeLa cell proliferation did not differ among different treatment periods. This suggests that tamoxifen may exert anti- proliferative effects at high concentration but does not have synergistic or antagonistic effects against estrogen on HeLa cell proliferation.

Human protein C (hPC) is a regulator of homeostasis, suggesting its potential use as a therapy for many disease states. Protein C is a zymogen of a serine protease that is activated by thrombin. Protein C, also known as autoprothrombin ⅡA and blood coagulation factor ⅪⅤ, is a zymogenic (inactive) protein, the activated form of which plays an important role in regulating blood clotting, inflammation, cell death and maintaining the permeability of blood vessel walls in humans and other animals. hPC is a 62 KD disulfide- linked heterodimer consisting of a 21 KD light chain and a 41 KD heavy chain which circulates as an inactive zymogen in plasma. In this study, we focus on generation of hPC transgenic mice. hPC transgenic mice were produced by using micro-injection method. The hPC cDNA was cloned into pBC1 vector under goat β-casein promoter. One-cell stage embryos microinjected were transferred to 24 recipient mice on day 1 of the estrus cycle. We screened 61 mice by the PCR. Four line transgenic mice were identified and confirmed expression of protein C gene in mammary gland and several organ. We also analyzed the expression of mRNA and protein through the northern blot and western blot in mammary gland of hPC transgenic mice. hPC was localized in the alveolar epithelial cell by immunohistochemistry. Now, we are collecting the milk from the 2 found lines. After then, we are checking the activity of hPC produced from mice milk.

20ɑ-hydroxysteroid dehydrogenase (20ɑ-HSD) enzyme converts progesterone into biological inactive steroid, thus playing a key role in the termination of pregnancy or estrus cycle and allowing parturition and ovulation to occur in most mammalian animals. However, function and regulation of this enzyme has not known well in primate reproductive physiology. We previously demonstrated the expression level and localization of the 20α-HSD in the reproductive tissues of macaque monkeys of pre-ovulation and pre-parturition period. Also, we amplified about 2005 bp 5'-flanking region from placenta genomic DNA and examined methylation pattern and promoter activity. In present study, we focus on the analysis of molecular characterization of the promoter region by using reporter assay systems. We constructed of deleted mutants (— 890 bp; HSF-2), (— 513 bp; XFD), (— 276 bp; Ap-1) and (— 72 bp; Sp-1) and each mutants were cloned into pGL3-basic vector. These deletion mutants were transfected into CHO cells and co-transfected with Sp-1 or Ap-1 transcription factor plasmids. Compared to — 890 bp and 513 bp promoter fragments alone, transcription activity increased when these constructs were co-transfected with Sp-1 and Ap-1 factor. However, for the absence Ap-1 factor binding site in 276 bp fragment activity dramatically decreased in both transfections. Next, we constructed of 306 bp fragment which is including of Ap-1 binding site and nucleotides converted mutants of the Ap-1 factor binding site. In this result, 306 bp fragment's transcription activity was high as wild type. However, the mutant activity which converted Ap-1 site’s all nucleotide was significantly decreased. These findings are confirmed by gel-shift assay examining Ap-1 binding site on the 20 α-HSD gene upstream region and expression of Ap-1 factor was determined by RT-PCR and Western blot in pre-parturition period placenta and CHO-K1 cell line. Our results indicate that Ap-1 site (— 281 → — 274) (5'-TGTCTCAT-3') plays a crucial role for monkey 20 α- HSD gene transcription.

This study was to analyse the usability of morphological evaluation of vitrified-thawed oocyte before somatic cell nuclear transfer (SCNT) using Oosight imaging system to show spindle. For the vitrification, in vitro matured bovine MII oocytes were treated by two-step freezing medium without (control group) or with 5 ug/ml cytochalasin-b (CCB group). In Exp. 1, after thawing, recovered oocytes in each treatment group were assessed by live image using Oosight imaging system or/and cytoskeletal protein image using immunostaining. In Exp. 2, in each treatment group the in vitro developmental potential of frozen-thawed bovine oocytes post evaluation using Oosight imaging system and then SCNT was investigated. The SCNT embryos were cultured in CR1aa medium supplemented with 10% FBS, 1 mM EGF and 1 mM IGF at 38.5 C in 5% O2 and 5% CO2 in air for 8 days. In Exp.1, the rates of in vitro survival, morphological good grade and spindle normality of CCB treatment group (91.1%, 54.2% and 55.5%) were better than those of control group (86.1%, 48.5% and 48.5%). After SCNT using vitrified-thawed oocyte, the rates of fusion, reconstructed embryos and blastocyst development were also high in CCB treatment group (66.6%, 36.4% and 3.0%) than control group (60.0%, 27.3% and 0%). These results demonstrated that the identification of morphological spindle image of the vitrified-thawed bovine oocytes using Oosight imaging system helps to predict the SCNT embryo quality.

SERPINB3 (also known Squamous cell carcinoma antigen 1, SCCA1) is involved in apoptosis, immune response, cell migration and invasiveness of cells. It has been investigated in various types of squamous cell carcinoma. Therefore we investigated the functional role of SERPINB3 gene in human epithelial ovarian cancer (EOC) using laying hens, the most relevant animal model. In 136 laying hens, EOC was found in 10 (7.4%). We compared the expression and localization of SERPINB3 using RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemistry, and SERPINB3 activation was detected in chicken and human ovarian cancer cell lines using immunofluorescence microscopy. Thereafter, we examined the prognostic value of SERPINB3 expression in patients with EOC by multivariate linear logistic regression and Cox’ proportional hazard analyses. In present study, SERPINB3 mRNA was induced in cancerous ovaries (p< 0.01), and it was only expressed in the glandular epithelium(GE) of cancerous ovaries of laying hens. SERPINB3 protein was localized predominantly to the nucleus of glandular epithelium in cancerous ovaries of laying hens, and it was abundant in the nucleus of both chicken and human ovarian cancer cell lines. In 109 human patients with EOC, 15 (13.8%), 66 (60.6%) and 28 (25.7%) of those patients showed weak, moderate and strong expression of SERPINB3 protein, respectively. Strong expression of SERPINB3 protein was a prognostic factor for platinum resistance (adjusted OR, 5.94; 95% Confidence Limits, 1.21-29.15). Therefore SERPINB3 may play an important role in ovarian carcinogenesis and be a novel biomarker for predicting platinum resistance and a poor prognosis for survival in patients with EOC. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010- 0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

The aim of this study was to investigate the correlation among bone mineral density(BMD), body composition and body circumference on 20's college women in Hwaseong. A total of 86 subjects were measured with BMD and body composition and body circumference. To evaluate the correlation between BMD and body composition, bone density and body weight, body mass index(BMI), lean body mass, muscle mass, fat mass and body fat mass were compared. The results of this study, weight was considered the strong correlation with BMD than the height and BMI seems to be greater significance rather than the lumbar spine and femur BMD. In addition, the relationship between body composition and BMD, lean body mass, muscle mass, body fat mass were the most relevant factors and BMD. The relationship between BMD and body circumference that have been difficult because of not enough previous studies but somewhat the study showed that association.

The purpose of this study was to analyze lower limb muscle activity and 3D motion analysis according to change foot arch height during walking. We selected 9 young and healthy people who have been normal foot. And we selected 7 young and healthy people who have been flatfoot. So, people were divided into 2 groups and walked platform during 2 minutes twice for checked by 3D motion analysis. These data were characterized by EMG measurements of three muscles( tibialis anterior, medial and lateral gastrocnemius) while they were walking. The collected data were analyzed by Independent t test using the SPSS statistics program(Ver 12.0). In foot arch change, there were no significant difference in three muscles 3D motion analysis also found that there were no significant difference in joint angles. In this study was to analyze lower limb muscle activity and 3D motion analysis according to change foot arch, but there were no significant difference in 6 muscles neither joint angles.

This study was carried out to identify how a self-stretching exercise program affects pain for each body area, pain relief and job satisfaction for care workers. 20 of 40 care workers with musculoskeletal symptom were randomly selected and participated a self-stretching exercise program consisting of 15 motions. The intervention was done five times or more per weeks for 8 weeks and 1 session lasted within 15 minutes. 'Musculoskeletal symptom survey table' of the Korea Occupational Safety and Health Agency(KOSHA) and JDI(Job Descriptive Index) was used for pain on the musculoskeletal symptom and job satisfaction. Survey were done twice before and after the program. The result of this study showed that self-stretching exercise program group(SSPG) relieved from pain significantly in the shoulders(p<.01) and lumbar(p<.05), comparing to the non selfstretching exercise program group(NSPG). Although no significant difference on variations in the JDI appeared in SSPG, the significant reduction appeared from the colleague relationship and organization in NSPG(p<.05). SSPG showed the significant increase on variations in JDI from the job and organization comparing to NSPG. Especially, the improvement on satisfaction for the organization was shown(p<.05). Accordingly, the self-stretching exercise program for care workers can be said to positively affect the overall pain relief and increase on the JDI.

Experiments were conducted in order to assess the healing effect of bee venom (BV) cream on full-thickness skin wounds in rabbits. BV cream was compared with silver sulfadiazine (SS) as a topical medicament against a control on experimentally created full-thickness wounds. Two wounds measuring 2 × 2 cm were created bilaterally (four wounds/rabbit) on the dorsolateral aspect of the trunk of seven New Zealand white rabbits. Wound treatments were evenly distributed on four sites, using a Latin square design. The contact layer of wounds was treated with physiological saline (control), SS cream, and BV cream over a period of 28 days. Assessment of wound healing was based on scab hardness, wound exudates, wound area, unepithelialized granulation tissue, and histopathological findings. Topical application of BV and SS creams to wounds resulted in reduced inflammation, debridement of necrotic tissue, and promoted granulation and epithelialization. Wound healing was faster, with statistical significance in BV and SS treatments, compared to the control (P<0.05). Treatment with BV evoked an anti-inflammation effect in a rabbit model. BV cream produced a wound healing effect similar to that of commercially available SS cream. Anti-inflammation effect as a topical treatment with BV cream appears to be better than that with SS cream. These results suggest that topical application of BV cream may be an alternative treatment for full-thickness skin wounds.

A novel strategy for the simultaneous reduction and functionalization of graphene oxide (G-O) was developed using polyallylamine hydrochloride (PAAH) as a multi-functional agent. The G-O functionalization by PAAH was carried out under basic conditions to catalyze the epoxide ring opening reaction of G-O with abundant amine groups of PAAH. We found that G-O was not only functionalized with PAAH but also reduced under the reaction condition. Moreover, the synthesized PAAH-functionalized G-O sheets were soluble in water and applicable to the synthesis of nanocomposites with gold nanoparticles.

This study examines impartiality of an EFL academic language rest and compares academic language abilities for Indo-European (IE) and non Indo-European (NIE) language groups. Multiple-sample confirmatory factor analysis with mean structures was conducted on the Test of English as a Foreign Language (TOEFL) scores. Findings indicate that partial measurement invariance was established for both groups. The two-factor measurement model (LC and non-LC) was found the best fitting for both the groups, and factor loadings were comparable. Also, even though two intercepts (vocabulary and reading) were different across both groups, the difference was not practically meaningful. Accordingly, it was concluded that impartiality of the TOEFL score interpretation for the two groups could be supported. Findings also showed that mean academic language abilities (LC and non-LC) were not significantly different between the two groups, which lead to the conclusion that the receptive academic language abilities arc not influenced by the difference in L1 or in the way of language education they received. The claims about decisions and consequences of the use of the TOEFL were also discussed.

The objective of this study was to determine the effects of E. coli isolated from porcine semen on sperm viability, motility, and semen pH. Semen samples were prepared using commercial extender, SeminarkPro (Noahbio Tech, Korea) that did not contain antibiotics. And 4 different levels of E. coli were artificially innoculated to semen with following concentrations; 4,000 of sperms with 1 of E. coli (T1), 400 with 1 (T2), 40 with 1 (T3), and 4 with 1 (T4). Semen samples were preserved at 17℃ for 5 days in semen storage box until analyzed by flowcytometer. Aliquots were subjected to measure the sperm viability (Live/Dead® stain), motility (mitochondrial function), and semen acidity (pH) from day 0 (day of semen collection) to day 5. Sperm motility and viability were significantly decreased (p<0.05) on day 0 (4 hrs after preservation at 17℃) in T3 and T4 compared to control groups and were significantly decreased (p<0.05) in all groups from day 3. Sample pH was acidic in T3 (6.90～6.86) and T4 (6.86～6.65) from day 3 to day 5 (p<0.05). On the other hand, sample pH was maintained 7.0～7.1 in control, T1, and T2 during the experimental period. Sperm motility and viability were significantly decreased from day 0 to day 5 compared to control in samples contaminated with E. coli above a value of 40:1 (20×106 sperm cells/ml : 5×105 cfu/ml). Even on day 1 in T4 and on day 3 in T3, semen pH was acidic probably due to the acidification of dead spermatozoa. These results suggest that E. coli contamination has a concentration-dependent detrimental effect on extended porcine semen quality.

The purpose of this study was to investigate the influence of handicraft activities on hand promptness and grasp in the elderly. Subjects were comprised of 14 senior citizens between the ages of 70-85, with 7 subjects in the experiment group and 7 in the control group. Subjects in the experiment group practiced various handicrafts twice a day, while those in the control group did not participate in any special activity. The Jebsen Taylor Hand Function Test was used to evaluate the results, while a dynamometer and pinch gauge were used to measure hand promptness and grasp. The 7 senior citizens in the experiment group were able to increase their hand promptness and grasping skills. Conclusively, handicrafts can help improve hand promptness and grasp in the elderly. Furthermore, the development and improvement of such skills can have a positive influence on the daily lives of senior citizens. Such skills are expected to improve the overall neuro-function in the elderly population.

The purpose of this study is to examine the changes in bone mineral density in long-stay patients of a geriatric hospital in relation to physical therapy, thereby providing basic data for preventing the onset or deterioration of osteoporosis in inpatients of geriatric hospitals or relevant facilities. The subjects of this study were 133 elderly patients who had been in H geriatric hospital in Yongin-si for more than four years. Their bone mineral density T-scores at admission and those after four years were measured to compare and analyze the changes in bone mineral density in relation to physical therapy. According to the results, the decrease in T-score for the group without physical therapy was greater by .40 than that for the group with physical therapy, indicating a statistically significant difference between the two groups(p<.01). Accordingly, physical therapy proved to be effective in preventing bone mineral density loss and maintaining the current density.

Urokinas type plasminogen activator (uPA) has been used as a therapeutic agent for treating human diseases such as thrombosis. Attempts to transgenically overexpress the uPA in animal bioreactors have been hampered due to side effects associated with this functional protein hormone on homeostasis. Recently, chicken has been emerged as a potential candidate for use as bioreactor to produce proteins of pharmaceutical importance. Since this species has low homology uPA sequence with mammals, we hypothesized that chicken could be used as a potential bioreactor for production of human uPA. In this study, using replication‐defective Murine Leukemia Virus (MLV)‐based retrovirus vectors encapsidated with Vesicular Stomatitis Virus G Glycoprotein (VSV‐G), we attempted to make transgenic chicken expressing human uPA (huPA). The recombinant retrovirus was injected beneath the blastoderm of non‐incubated chicken embryos (stage X, at laying). After 21 days of incubation (at hatching), all of the 38 living chicks that assayed, were found to express the vector‐encoded huPA gene in various organs and tissues, which was under the control of the Rous Sarcoma Virus (RSV) or Cytomegalovirus (CMV) promoter. Using specific primer set for huPA, PCR and RTPCR analyses of gDNA isolated from these samples demonstrated these chickens were transgenic for huPA. Furthermore, successful germ line transmission of huPA transgene was confirmed and next generation whole body huPA transgenic chickens were also produced. We also assayed huPA protein titer in blood (17.1 IU/ml) and eggs (4.4 IU/ml) of whole body huPA transgenic chicken. Thus, our results demonstrated that chicken could be used as bioreactors to produce huPA.

Somatic cell nuclear transfer (SCNT) is an efficient technique which has been successfully applied to developmental biology, and resulted in the production of offspring from various species. It offers many opportunities in basic and medical research as well as endangered species preservation. On the other hand, embryonic stem (ES) cells are useful research tools for genetic engineering and developing disease models. In previous study, we established bovine IVF embryo derived ES cell line which can be grow indefinitely as undifferentiated cell state. In this study, we compared the effect of two different age cells (bovine ES cell; JNU-ibES-05 or adult ear fibroblast cell) on in vitro developmental potential of bovine SCNT embryo. To produce SCNT embryos, the ES cells or somatic cells were dissociated and transferred into enucleated MⅡ oocytes, and cleaved reconstructed embryos were cultured in CR1aa medium containing 10% FBS, 1 ug/ml epidermal growth factor (EGF) and 1 ug/ml insulin growth factor (IGF) for 8 days. In the result, blastocyst development rate was similar between ES cell treatment group and somatic cell treatment group, 27.7% (10/36) and 28.9% (11/ 38), respectively. However, there was particular difference in development speed from day 5 post SCNT, blastocyst expanding was 1 day faster in ES cell group than in somatic cell group. This difference was analyzed by semi-quantitative RT-PCR using pluripotency, growth and cell cycle gene markers. These results demonstrated that SCNT embryo using ES cell as a donor cell has better growth potential than somatic cell, and it will be a useful tool for a transgenic animal production.