The human ELAV(embryonic lethal abnormal vision)-like protein HuR stabilizes a certain group of cellular mRNAs that contain AU- ri ch elements in theil‘ 3’• untranslated region, Dysregulation of mRNA s tability may be relevant in tumor biology and may lead to abnormal expression of several proteins in malignant tumors, The aim of this study is to identify the differentially expressed proteins according to the functi onal activi ty of HuR Methods : We used stabl e expression of small interfering RNA(siRNA) of HuR gene to inhibit the expression of HuR in human ora l car cinoma cell lines‘ KB cell line and YD10B cell line‘ and compared the proteomic changes between s iRNA-treated and cont rol cell line using two-dimensional gel electrophoresis , Flow cytometry caliber scan(FACS) was employed to investigate the effects of HuR on cell apoptosis and proliferation , Results: Seventeen differentially expressed proteins between the two cell lines were identified by electrospray ioruzati on quadrupole time-of- fl ight mass spectrometry(ESI-Q-TOF MS) and database searching, Among them there are eleven proteins which are significant ly up- regulated in siRNA- treated cell line, which include heat shock protein 10(influencing nucl eocytoplas rnic transpo 다， cell dedifferentiation , and inhibition of apoptosis) , keratin 19(basal cell differ ent iat ion) ‘ and nucleoside diphosphate kinase B(G protein activator), etc Enolase 1 and Ml of pyruvate kinase a re the representatives of signifi cantly down-regulated proteins in siRNA-treated cell 11l1e Conclusion : Our data suggest that HuR participate in mRNAs stability of proteins that have the counter effects in the carcinogenesis of oral ca ncer , And the functiona l proteomics a re needed to elucidate the detailed interactions between HuR and t hese molec ules

The present study investigated the effects of follicle stimulating hormone (FSH) and human chorionic gonadotrophin (hCG) on the nuclear maturation of canine oocytes. Oocytes were recovered from mongrel female ovaries in various reproductive states; follicular, luteal or anestrous stage. Oocytes were cultured in serum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.5, 1.0 or 10 IU) or hCG (Exp. 2: 0, 0.5, 1.0 or 10 IU) or both (Exp. 3: 1 IU FSH + 1 IU hCG) for 72 hr to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with 1.9 μM. Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. More (p<0.05) oocytes matured to MII stage when follicular stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the control, 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (p<0.05) maturation rate to MII stage was observed in follicular stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG supplemented groups (0 to 1.5%). However, the combination of FSH and hCG did not improve the nuclear maturation rate of canine oocyte (2.4 %) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to MII stage.

Transforming growth factor-β (TGF-β) has been shown to have a positive effect on in vitro fertilization (IVF) and has been reported to stimulate meiosis at follicular level in variety of species. The study was designed to determine the expression patterns of TGF-β1, TGF-β receptors type Ⅰ, Ⅱ and Smads gene in bovine oocytes and embryos. TGF-β1 and their receptors were observed in the unfertilized oocytes. TGF-β1 and type Ⅱ receptor were not expressed at the blastocyst stage, however, only type I receptor was exclusively observed at the same stage. The blastocyst stage, in particular, showed high levels of mRNA expression patterns containing a TGF-β type Ⅰ receptor. The mRNA expression pattern of Smad 2 at all stages of embryonic development was similar in all respect with TGF-β1 type I receptor. On the contrary, Smad 3 and 4 were expressed with high and low level mRNA at the blastocyst stage. In conclusion, it is suggested that TGF-β signaling may be regarded as an important entity during the preimplantation embryo development.

Floral scents and metabolites from cut flowers of 14 peony cultivars (Paeonia lactiflora Pall.) were analyzed to discriminate different cultivars and to compare the Korean cultivar with the other cut peonies imported to Korea using electronic nose (E-nose) and Fourier transform infrared (FT-IR) spectroscopy combined with multivariate analysis, respectively. Principal component analysis (PCA) and discriminant function analysis (DFA) dendrogram of peony floral scents were not precisely same but there were 3 groups including same cultivars. PCA and partial least squares-discriminant analysis (PLS-DA) dendrograms of peony metabolites showed that different cut peony cultivars were clustered into two major groups including same cultivars. Fragrance pattern of Korean ‘Taebaek’ was classified to same group with ‘Jubilee’ on the PCA and DFA results and its metabolite pattern was clearly discriminated by the PCA and PLS-DA compared to the other cultivars. These results show that the 14 peony cut flowers could be discriminated corresponding to their chemical relationship and the metabolic profile of Korean ‘Taebaek’ has distinctive characteristics. Furthermore, we suggest that these results could be used as the preliminary data for breeding new cut peony cultivars and for improving the availability of Korean cut peony in cosmetic industry.

본 연구는 LPS로 유도된 RAW 264.7 세포에서 다양한 약용식품으로 사용되고 있는 올리브 잎과 가지 추출물의 항염증 효과를 확인하였다. 올리브 잎과 가지 추출물은 각각 RAW 264.7 세포에 대하여 세포독성을 나타내지 않았고, LPS 자극에 의한 NO 및 PGE2 생성을 농도 의존적으로 억제했다. 또한, 올리브 추출물은 LPS 자극으로 분비된 TNF-α, IL-1β 및 IL-6의 전염증성 cytokine의 분비량을 억제하였으며, 특히 200 μg/mL 농도에서 올리브 가지 추출물이 잎 추출물 보다 IL-6를 억제하는 것으로 나타났다. 대표적인 염증 관련 신호 전달 경로 인자인 iNOS 및 COX-2의 발현을 검토한 결과 올리브 추출물은 iNOS의 발현을 농도의존적으로 현저히 감소시키는 것으로 관찰되었으나, 각각의 올리브 추출물이 COX-2 발현에는 영향을 미치지 않은 것으로 관찰되었다. 이와 같은 결과는 올리브 각 부위별 추출물은 모두 iNOS 및 NO 조절 경로를 조절하는 것으로 사료되나 iNOS 및 COX-2 단백질 발현은 병립적이지 않을 수 있음을 제시하고 있다. 본 연구 결과로 올리브 추출물이 독성과 부작용이 적은 항염증 효능을 가진 기능성 화장품 소재로써 개발 가능성이 있다고 사료된다.

In this study, we elucidated the molecular mechanism of silymarin by which silymarin may inhibits cell proliferation in human colorectal cancer cells in order to search the new potential anti-cancer target associated with the cell growth arrest. Silymarin reduced the level of c-Myc protein but not mRNA level indicating that silymarin-mediated downregulation of c-Myc may result from the proteasomal degradation. In the confirmation of silymarin-mediated c-Myc degradation, MG132 as a proteasome inhibitor attenuated c-Myc degradation by silymarin. In addition, silymarin phosphorylated the threonine-58 (Thr58) of c-Myc and the point mutation of Thr58 to alanine blocked its degradation by silymarin, which indicates that Thr58 phosphorylation may be an important modification for silymarin-mediated c-Myc degradation. We observed that the inhibition of ERK1/2, p38 and GSK3β blocked the Thr58 phosphorylation and subsequent c-Myc degradation by silymarin. Finally, the point mutation of Thr58 to alanine attenuated silymarin-mediated inhibition of the cell growth. The results suggest that silymarin induces the cell growth arrest through c-Myc proteasomal degradation via ERK1/2, p38 and GSK3β-dependent Thr58 phosphorylation.

Background : Arctii Radix (the dried roots of Arctium lappa L., AR) is used in traditional medicine to treat oxidative stress related diseases including cancer. Therefore, this study focuses on the antioxidant potential of AR as extraction solvents. Methods and Results : To increase the extraction amount of active ingredient, the AR were extracted by ethanol (ARE) and water (ARW). In order to determine active ingredient content of AR, we were carried out total polyphenolic (TPC) and flavonoid content (TFC) analyses. As a result, TPC (47.74 ± 1.02 g․GAE/㎏ extract) and TFC (19.34 ± 0.30 g․CTE/㎏ extract) of ARE were found significantly higer as compared to ARW. The IC50 values based on the DPPH (59.00 ± 3.25 ㎍/㎖), ABTS (93.20 ± 1.30 ㎍/㎖), ROS (57.78 ± 3.44 ㎍/㎖) and ONOO- (14.56 ± 1.24 ㎍/㎖) for ARE were generally stronger showing potential antioxidant properties compared to ARW. Conclusion : Data from results revealed Arctii Radix ethanol extracts act as an antioxidant agent due to its free radical scavenging activity.

High yield is the most important trait in various agricultural characteristics. Many approaches to improve yield have been tried in conventional agricultural practice and recently biotechnological tools employed for same goal. Genetic transformation of key genes to increase yield is one way to overcome current limitation in the field. We are producing transgenic soybean plants through high efficient transformation method by introducing all gene member with AT-hook binding domain, hoping to obtain manageable delay of senescence. Many transgenic soybean plants are growing in greenhouse and GMO field, and will be evaluated their senescence and any association with yield increase.

Soybean is a crop of importance economically and nutritionally in many parts of the world. Thanks to many new genes brought from genomic research, It is possible to introduce various candidate genes through genetic transformation to see the performance of the genes in field. In our lab, soybean transformations have been tried for last 10 years to probe the possibility of traits improvement by transformation of new gene into soybean. For this purpose, three different genes were transformed into Korean soybean variety, Kwangan. First, the gene that controls early flowering of plant was transformed into Kwangan. Second, a candidate gene for soybean mosaic virus (SMV) resistance was transformed to produce transgenic plants. Third, another candidate gene for drought tolerance was transformed. All the transgenic plants from three genes transformation were produced for their gene insertion and their expression using PCR, qRT-PCR. Further analysis including harvesting seeds is currently undertaken.

To understand molecular mechanisms underlying adaptation of plant cells to saline stress and stress memory, we developed Arabidopsis callus suspension-cultured cells adapted to high salt. Adapted cells to high salt exhibited enhanced tolerance compared to control cells. Moreover, the salt tolerance of adapted cells was stably maintained even after the stress is relieved, indicating that the salt tolerance of adapted cells was memorized. Salt-adapted and stress memorized cells were densely aggregated and formed multi-layered cell lump. Cell morphology analysis using transmission electron microscopy indicated that cell wall thickness of salt-adapted cells was significantly induced compared to control cells. In order to characterize metabolic responses of plant cells during adaptation to high salt stress as well as stress memory, we compared metabolic profiles of salt-adapted and stress-memorized cells with control cells by using NMR spectroscopy. A principle component analysis showed clear metabolic discrimination among control, salt-adapted and stress-memorized cells. Compared with control cells, metabolites related to shikimate metabolism such as tyrosine, and flavonol glycosides, which are related to protective mechanism of plant against stresses were largely up-regulated in adapted cell lines. Moreover, coniferin, a precursor of lignin, was more abundant in salt-adapted cells than control cells. The results provide new insight into metabolic level mechanisms of plant adaptation to saline stress as well as stress memory.

To identify novel signaling components involved in regulation of plant responses to phosphate (Pi) starvation, we screened an Arabidopsis T-DNA activation tagging library for mutants with altered Pi-starvation responses. Here, we report the identification and characterization of novel activation-tagged mutant involved in Pi starvation signaling in Arabidopsis. The hpd (hypersensitive to Pi deficiency) mutant exhibits enhanced phosphate uptake and altered root architectural change under Pi starvation compared to wild type. Expression analysis of auxin-responsive DR5::GUS reporter gene in hpd mutant indicated that both auxin biosynthesis and auxin translocation under Pi starvation are suppressed in hpd mutant plants. Impaired auxin translocation in roots of hpd mutant was attributable to abnormal root architecture changes in Pi starvation conditions. Mis-regulation of auxin translocation in hpd mutant was further confirmed by analysis of expression patterns of auxin efflux carrier proteins, PIN-FORMED (PIN) 1, 2, and 3 fused with GFP. Not only expression levels but also expression domains of PIN proteins were altered in hpd mutant in response to Pi starvation. Molecular genetic analysis of hpd mutant revealed that the mutant phenotype is caused by the lesion in ENHANCED SILENCING PHENOTYPE4 (ESP4) gene whose function is proposed in mRNA 3’-end processing. The results propose that mRNA processing plays crucial roles in Pi homeostasis as well as developmental reprograming in response to Pi deprivation in Arabidopsis.

The flower buds of Sophora japonica L (SF), as a well-known traditional Chinese medicinal herb, have been used to treat bleeding-related disorders such as hematochezia, hemorrhoidal bleeding, dysfunctional uterine bleeding, and diarrhea. However, no specific anti-cancer effect and its molecular mechanism of SF have been described. Thus, we performed in vitro study to investigate if treatment of SF affects activating transcription factor 3 (ATF3) expression and ATF3-mediated apoptosis in human colorectal cancer cells. The effects of SF on cell viability and apoptosis were measured by MTT assay and Western blot analysis against cleaved poly (ADP-ribose) polymerase (PARP). ATF3 activation induced by SF was evaluated using Western blot analysis, RT-PCR and ATF3 promoter assay. SF treatment caused decrease of cell viability and increase of apoptosis in a dose-dependent manner in HCT116 and SW480 cells. Exposure of SF activated the levels of ATF3 protein and mRNA via transcriptional regulation in HCT116 and SW480 cells. Inhibition of extracellular signal-regulated kinases (ERK) 1/2 by PD98059 and p38 by SB203580 attenuated SF-induced ATF3 expression and transcriptional activation. Ectopic ATF3 overexpression accelerated SF-induced cleavage of PARP. These findings suggest that SF-mediated apoptosis may be the result of ATF3 expression through ERK1/2 and p38-mediated transcriptional activation.

Mannitol is commonly used to reduce intracranial and intraocular pressures and to prevent dialysis-disequilibrium syndrome. However, intravenous mannitol infusion in various cases has the potential to result in acute kidney injury (AKI). We present a case of mannitol-induced AKI that developed after low dose mannitol infusion and resulted in recovery after hemodialysis. A 66-year-old woman was admitted to the hospital with a diagnosis of left middle cerebral artery infarction. On hospital day 5, cerebral edema was observed on a follow-up MRI. D-mannitol 35 g was given intravenously every 8 hours. Four days later, serum creatinine levels were elevated from 1.2 mg/dL to 3.5 mg/dL. The serum osmolal gap was found to be 52.4 mosm/kg H2O and urine output was reduced from 2.78 mL/kg/h to 0.69 mL/kg/h over three days. Hemodialysis over 2 hours was performed and renal function subsequently improved to baseline function. A potential risk of AKI exists even with low dose mannitol infusion in patients with advanced age, underlying renal impairment, and concomitant use of nephrotoxic agents. Mannitol-induced AKI may be rapidly reversed by short-term hemodialysis.