Background: Coating a culture plate with molecules that aid in cell adhesion is a technique widely used to produce animal cell cultures. Extracellular matrix (ECM) is known for its efficiency in promoting adhesion, survival, and proliferation of adherent cells. Gelatin, a cost-effective type of ECM, is widely used in animal cell cultures including feeder-free embryonic stem (ES) cells. However, the optimal concentration of gelatin is a point of debate among researchers, with no studies having established the optimal gelatin concentration. Methods: In this study, we coated plastic plates with gelatin in a concentrationdependent manner and assessed Young’s modulus using atomic force microscopy (AFM) to investigate the microstructure of the surface of each plastic plate. The adhesion, proliferation, and differentiation of the ESCs were compared and analyzed revealing differences in surface microstructure dependent on coating concentration. Results: According to AFM analysis, there was a clear difference in the microstructure of the surface according to the presence or absence of the gelatin coating, and it was confirmed that there was no difference at a concentration of 0.5% or more. ES cell also confirmed the difference in cell adhesion, proliferation, and differentiation according to the presence or absence of gelatin coating, and also it showed no difference over the concentration of 0.5%. Conclusions: The optimum gelatin-coating for the maintenance and differentiation of ES cells is 0.5%, and the gelatin concentration-mediated microenvironment and ES cell signaling are closely correlated.

Lipopolysaccharide (LPS) is an endotoxin factor present in the cell wall of Gram-negative bacteria and induces various immune responses to infection. Recent studies have reported that LPS induces cellular stress in various cells including oocytes and embryos. Melatonin (N-acetyl-5-methoxytryptamine) is a regulatory hormone of circadian rhythm and a powerful antioxidant. It has been known that melatonin has an effective function in scavenging oxygen free radicals and has been used as an antioxidant to reduce the cytotoxic effects induced by LPS. However, the effect of melatonin on LPS treated early embryonic development has not yet been confirmed. In this study, we cultured mouse embryos in medium supplemented with LPS or/and melatonin up to the blastocyst stage in vitro and then evaluated the developmental rate. As a result of the LPS-treatment, the rate of blastocyst development was significantly reduced compared to the control group in all the LPS groups. Next, in the melatonin only treated group, there was no statistical difference in embryonic development and no toxic effects were observed. And then we found that the treatment of melatonin improved the rates of compaction and blastocyst development of LPS-treated embryos. In addition, we showed that melatonin treatment decreased ROS levels compared to the LPS only treated group. In conclusion, we demonstrated the protective effect of melatonin on the embryonic developmental rate reduced by LPS. These results suggest a direction to improve reproduction loss that may occur due to LPS exposure and bacterial infection through the using of melatonin during in vitro culture.

본 연구는 원통형 종이포트를 활용한 토마토 육묘시, 염스트레스를 활용하여 고온기 도장 억제가능성을 검토하기 위하여 수행되었다. 시험구는 K2SO4, KCl과 KH2PO4을 각 5, 10 dS·m-1로 처리하였고, 또한, 토마토 모종에 고염도의 칼륨을 처리하여 수분 및 저온스트레스 환경에서의 적응성 및 생존성을 조사하였다. 조사결과, 처리 농도가 높아질수록 지상·지하부 건물중, 옆면적, 순동화율 (NAR)이 감소하고, 경경과 충실도는 증가하였다. 수분 스트레스 처리 이후, 대조구는 심한 위조현상을 보였지만, KCl처리구는 양호하였다. 상대수분함량은 대조구에서 23%, KCl처리구에서 8% 감소 하였다. 또한, 대조구에 비하여 KCl 처리구는 저장시(9, 12 및 15°C) 모종의 손상 비율이 낮았다. 이와 같은 결과로 보아, KCl과 같은 고농도의 칼륨 처리가 원통형 종이포트 토마토 육묘의 도장 억제에 효과적이며 환경 스트레스 내성을 향상시키는 것으로 판단된다.

Endoplasmic reticulum (ER) stress is well known as a suppressor in osteoblast differentiation and activating transcription factor 3 (ATF3) could be induced by a various extracellular signals including cytokines, hormones, DNA damage. Up to date, although the role of ATF3 have been studied, the function of ATF3 in osteoblast differentiation is still not clear yet. Our study showed that expression level of ATF3 could be incresed by tunicamycin which is ER stress inducer in preosteoblasts. BMPs, which are secreted by osteoblasts, can be important regulators in osteogenic differentiation. The stress-responsive transcription factor ATF3 is a negative regulator of osteoblast differentiation in MC3T3-E1 cells. In this study, we verified that BMP2-stimulated osteoblast differentiation could be inhibited by over-expressed ATF3 through regulating alkaline phosphatase (ALP) expression and activation.

선박이나 컨테이너로 수출입되는 곡류 및 박류는 검역용 훈증제인 메틸브로마이드(이하 MB)와 인화수소(이하 PH3)를 사용하는데, 처리 온도/수용비/선적방식에 따라 충분한 배기시간을 필요로 한다. 현장에서 배기시간 미설정으로 검역관 및 방제기술자에게 훈증제 TLV(Threshold Limit Value)-TWA(Time Weighted Average) 기준 이상의 농도에 노출될 위험이 있으므로, 소독 전, 후 작업자 안전을 고려하여 각 훈증제 별 작업자 안전기준에 적합한 배기시간이 설정되어야 한다. 따라서, 실내 훈증상에서 곡류(쌀, 대두) 및 박류(대두박, 주정박)를 대상으로 현행 소독처리기준으로 소독 후 탈착되는 가스농도를 측정해 MB 1 ppm, PH3 0.3 ppm 수준 이하로 감소되는 배기시간을 조사하였다. 조사결과 TLV-TWA 기준이하의 수준으로 감소되는데 필요한 배기시간은 주정박/대두박/대두에 MB가 처리된 경우는 모두 30시간, 쌀에 PH3가 처리된 경우는 30분이 소요되는 것을 확인하였다.

The shortage of human organs for transplantation has induced the research on the possibility of using animal as porcine. However, pig to human transplantation as known as xeno-transplantation has major problem as immunorejection. Recently, the solutions of pig to human xenotransplantation are commonly mentioned as having a genetically modification which include alpha 1, 3 galatosyl transferase knockout (GTKO) and immune-suppressing gene transgenic model. Unfortunately, the expression level of transgenic gene is very low activity. Therefore, development of gene overexpression system is the most urgent issue. Also, the tissue specific overexpression system is very important. Because most blood vessels are endothelial cells, establishment of the endothelial-specific promoter is attractive candidates for the introduction of suppressing immunorejection. In this study, we focus the ICAM2 promoter which has endothelial-specific regulatory region. To detect the regulatory region of ICAM2 promoter, we cloned 3.7 kb size mini-pig ICAM2 promoter. We conduct serial deletion of 5' flanking region of mini-pig ICAM2 promoter then selected promoter size as 1 kb, 1.5 kb, 2 kb, 2.5 kb, and 3 kb. To analyze promoter activity, luciferase assay system was conducted among these vectors and compare endothelial activity with epithelial cells. The reporter gene assay revealed that ICAM2 promoter has critical activity in endothelial cells (CPAE) and 1 kb size of ICAM2 promoter activity was significantly increased. Taken together, our studies suggest that mini-pig ICMA2 promoter is endothelial cell specific overexpression promoter and among above all size of promoters, 1 kb size promoter is optimal candidate to overcome the vascular immunorejection in pig to human xenotransplantation.

CREBZF (cAMP-response element binding protein zhangfei) is a member of ATF/CREB family, and which regulates various cellular functions by suppressing major factors with direct interaction. In this study, we have examined the expression of CREBZF on mouse endometrium during uterus estrous cycles and estrogen (E2) treatment. In uterus, CREBZF mRNA expression was higher than other organs and mRNA and protein of CREBZF was increased in proestrus phase and decreased in estrus phase. The expression of CREBZF in 3-weeks old mouse uterus was reduced by E2 injection in endometrium. In addition, the expression of progesterone receptor, a marker of E2 in ovariectomized mice was found to be strongly expressed in stroma, while CREBZF was only expressed in epithelium. Also, we conformed that E2-suppressed CREBZF was restored by co-injection of ICI 182,780, an estrogen receptor antagonist. Overall, these results suggest that CREBZF is regulated by estrogen and involved in ER signaling pathway in mouse uterus.

Cyclic AMP-response element binding protein zhangfei (CREBZF), a member of ATF/CREB (activating transcription factor/ cAMP response element binding protein) family, regulates numerous cellular functions and development of cells by interacting transcription factors. This study discovered the expression pattern of CREBZF in seminiferous tubule of testes during the postnatal development of mice. In testis, CREBZF mRNA expression was the highest among other organs. Immunofluorescence analyses showed that the CREBZF was specifically expressed on spermatocyte but not in spermatogonia and Sertoli cells in seminiferous epithelium of mouse testis. Semi-quantitative polymerase chain reaction (PCR) analysis showed that CREBZF transcript level was significantly elevated during postnatal development of mouse testis. Confocal imaging analysis indicated that the protein expression of CREBZF in seminiferous tubule remained low until postnatal day (PD) 14, and was dramatically increased in PD 21. Interestingly, only one type of the spermatocyte expressed CREBZF specifically among SCP3-positive spermatocytes. Taken together, these results suggest that CREBZF may be novel putative marker of the spermatocyte and regulate meiosis during postnatal development of mice.

Spermatogonial stem cells (SSCs; also known as Asingle [As] spermatogonia in mice) divide to self-renew or to produce progenitor cells known as Apaired(Apr) spermatogonia in basal compartment of seminiferous tubules of mammalian testis. These characterized cells are the finally differentiated product of a developmental process referred to as “spermatogenesis.” In the development of SSCs it is critical to maintain a balance between self-renewal and differentiation. because an excess of either process will lead to infertility. these two processes are tightly controlled by intrinsic signals of SSCs and extrinsic signals from the microenvironment, known as the SSC niche. The SSC niche is formed by Sertoli cells, the only somatic cells found inside the seminiferous tubules. The WNT/β-catenin pathway is known to regulate Sertoli cell functions critical to their capacity to support spermatogenesis in the postnatal testis, but The mechanisms and factors of the pathway are not well known. We found a factor TLE3 (Transducin Like Enhancer Of Split 3). The transcriptional co-repressor TLE family is known to function as transcription co-repressors within the context of Wnt signaling by interacting with histone deacetylase HDAC2. We examined the expression level of TLE3 in various mouse tissues. As a result of RT-PCR, TLE3 showed significantly higher expression in testis than that in other tissues. Immunofluorescent analysis revealed that TLE3 and HDAC2 expression are differentially regulated in the mouse testis during postnatal development. In adult testis, TLE3 and HDAC2 were co-expressed in Sertoli cells. TLE3 and HDAC2 protein are also located in nucleus in mouse TM4 Sertoli cells. Taken together, TLE3 may play a role in regulating WNT/β-catenin pathway via interaction with HDAC2 in Sertoli cell. Futher studies are needed to look into factors that regulated by siTLE3 in Sertoli cell and interated with TLE3 in WNT/β-catenin pathway.

The Hippo signaling pathway is essential for regulating proliferation, differentiation, and apoptosis in mammalian cells. Hippo signaling pathway exists in most body tissues and organs, where it controls the size of organs and tissues by keeping cell growth in check and promoting cell death as needed. It has been reported that the members of Hippo signaling pathway are highly expressed in mammalian ovaries and uteri. However, the regulatory mechanism of this pathway in the uterus during estrous cycle regulation remains unclear. Serine/Threonine Protein Kinase 4 (STK4, also known as MST1, a homolog of Hippo in Drosophila) is a major factor of Hippo signaling pathway. However STK4 in the mouse uterus has not yet been examined. The purpose of our study was to determine the expression of STK4 during the estrous cycle and regulation by estrogen in the mouse uterus. We found that STK4 was dynamically expressed in uterine endometrium during the estrous cycle. STK4 highly expressed at the estrus, diestrus, and were found to dramatically decrease as it progressed to the proestrus, metestrus stage of uterus during the estrous cycle. Expression of STK4 was dominant in glandular epithelial and luminal epithelial of proestrus, estrus, and diestrus stage, whereas in metestrus stage, expression of gene intensity was faint. Estrogen or estrogen receptor antagonist ICI 182,780 treatment, in ovariectomized mouse uterus, Expression of STK4 and its downstream genes were increased by estrogen. Our results show that the Hippo signaling pathway is estrogen-dependent in the mouse uterus. These informations will give us on sights to understand uterine dynamics during the estrous cycle.

This study aimed to investigate the effect of different light-emitting diode (LED) on the stress response of workers before and after a flower arrangement (FA) activity. Participants were 16 office workers who performed FA under 7 out of 8 LED lighting conditions: darkness, white (at correlated color temperature of 4,500 K), red, blue, green, magenta, cyan, and yellow; no FA is performed in the darkness condition. To identify significant differences among the different lighting conditions, the heart rate (HR), blood pressure (BP), heart rate variability (HRV) consisting of standard deviation of the normal-normal intervals (SDNN), total power (TP), ratio of the low and high frequency (LF/HF), and blood pressure were measured before and after FA. The salivary cortisol level was analyzed immediately after FA. The correlations among HR, HRV, and cortisol showed that the blue and green lights positively affected stress-responses and caused considerable reduction of stress. However, there was only a minor significant difference among other lighting colors, although individual lights may impact one or more parameters. These findings reflect that the methodologies and results of this study can be effectively used to achieve a highly active horticultural therapy with the constructive assistance of LED lighting colors.

The purpose of this study was to investigate the awareness of Daegu citizens on urban agriculture. A questionnaire was conducted for 15 days from May 9, to May 25, 2015, and a sample group consisting of 328 residents of Daegu. The cognitions of awareness to effects and preferences of urban agriculture were analyzed. The cognition level of urban agriculture is in a very low state, the level of satisfaction in experienced groups were high. In addition, intentions of participation in the future were very high. The potential capability of urban agriculture is very strong. In motivation and purpose of urban agriculture activities, such as ‘Hobby and leisure activities’, ‘Safe food production’ were shown as high frequency. In cognition towards the effects of urban agricultural activity, ‘Safe agricultural products’, and ‘Change in dietary life’ were shown to be in high points of agreement. Furthermore, in preferences of the type and activities, ‘Rooftop garden’, ‘Learning/education type home garden’, ‘Home garden utilizing the pieces of small land’, ‘Box home garden’, and ‘Personal weekend farm’ were shown to be highly preferred types. Within the effects of urban agriculture, 4 factors were categorized; ‘Negative effects of urban agriculture’, ‘Positive effects of urban agriculture’, ‘Environmental Improving effects’, and ‘Benefits effects’. In addition, 3 factors are prevalent in type and activities of urban agriculture, ‘Participations of experiencing/education programs’, ‘Cultivation of rental home gardens’, and ‘Lifestyle home garden type’. Conclusively, although the situation of urban agriculture is in the beginning stages, the potential capability of urban agriculture is great. In the future, diverse types of hardware and software programs, such as diverse education/experiencing programs and information supporting programs, will both be required for development