• Biomedical Science Letters
• /
• v.2 no.2
• /
• pp.231-240
• /
• 1996

Human blood clotting (coagulation) factor 9 cDNA which codes for 461 amino acid has been cloned by screening human fetal liver cDNA library using PCR. This 1.4 kb cDNA spanning from the ATG initiation codon to the TAA termination codon was cloned into bacterial .expression vector pGEX-2T, generating pGEX-F9 plasmid. The plasmid pGEX-F9 expresses about 73 kDa GST (Glutathione S-transferase)-Factor 9 fusion protein when introduced into E. coli. Western blot analysis using polyclonal antibody raised against human factor 9 confirmed this fusion protein contains factor 9 protein. The level of GST-factor 9 expression was about 20% of total protein and the purification of fusion protein was efficiently achieved by using GST agarose bead based on one step purification protocol.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.10
• /
• pp.1589-1603
• /
• 2018

Twenty analogs of $[Orn^6,D-Ala^9]{\alpha}-factor$ were synthesized and assayed for their biological activities: seven analogs of $[Orn^6,X^9]{\alpha}-factor$, seven analogs of $[X^6,D-Ala^9]{\alpha}-factor$, five analogs of $[X^5,X^6,D-Ala^9]{\alpha}-factor$, and native ${\alpha}-factor$ (X = amino acids). Their biological activities (halo, gene induction, and affinity) were measured using S. cerevisiae Y7925 and LM102 and compared with those of native ${\alpha}-factor$ (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. $[Dap^6,D-Ala^9]{\alpha}-factor$ with weak halo activity (10%) showed the highest receptor affinity (> 230%) and the highest gene induction activity (167%). $[Arg^6,D-Ala^9]{\alpha}-factor$ showed the highest halo activity (2,000%). The number of active binding sites per cell (about 20,000 for strain LM102) was determined using a newly-designed fluorescence-based detector, $[Arg^6,D-Ala^9]{\alpha}-factor-Edan$, with high sensitivity (12,500-fold higher than the absorption-based detector $[Orn^6]{\alpha}-factor-[Cys]_3$).

• Korean Journal of Construction Engineering and Management
• /
• v.12 no.5
• /
• pp.127-136
• /
• 2011

The purpose of this study is analyzed the importance ratio factors of sales price for apartment building construction projects. For this Study, Nine apartment buildings that sold in Seoul, Gyenggi, Chuncheong is selected. The study suggested eight influence factors through interview with experts, research related references, and analysis of cases. Based on those factors, Importance analysis is performed by using AHP method. The result of this study are as follows: 1) Influence factors of sales price are drawn as cost, site, housing, transportation, region, ground, policy, investment factor. 2) In the case of Seoul, The priority order of price influence factors consist of investment factor(16.0%), policy factor(14.6%), and cost factor(13.9%). In the case of Gyenggi, The priority order of sales price influence factors consist of region factor(15.9%), transportation factor(13.8%), and investment factor(13.4%). In the case of Chungcheong, The priority order of sales price influence factors consist of region factor(17.1%), transportation factor(16.4%), and housing factor(14.9%).

• Bulletin of the Korean Chemical Society
• /
• v.28 no.9
• /
• pp.1613-1614
• /
• 2007

• Bulletin of the Korean Chemical Society
• /
• v.29 no.9
• /
• pp.1860-1862
• /
• 2008

• Bulletin of the Korean Chemical Society
• /
• v.35 no.5
• /
• pp.1365-1374
• /
• 2014

Thirteen analogs of tridecapeptide ${\alpha}$-factor (WHWLQLKPGQPMY) of Saccharomyces cerevisiae with C- or N-terminal Trp extension and isosteric replacement by Aib at position 8 and 11, Trp at position 13, D-Ala at position 9, and Orn and Glu at position 6 were synthesized and assayed for their biological activity. Receptor binding assay was carried out using our newly developed spectrophotometric method with detector peptide 14. C- or N-terminal extended analogs, ${\alpha}$-factor-$[Trp]_n$ (n =1-5) 1-5 and $[N-Trp]_1$-${\alpha}$-factor 6, were all less active than native ${\alpha}$-factor and gradual decreases in both activity and receptor affinity were observed with greater Trp extension. Trp-substituted analog at position 13, $[Trp^{13}]{\alpha}$-factor 7, exhibited about 2-fold reductions in both activity and receptor affinity. Aib-substituted analogs, $[Aib^8]{\alpha}$-factor 8 and $[Aib^{11}]{\alpha}$-factor 9, showed 5- to 10-fold reduction in activity as well as 3-fold reduction in receptor affinity compared to native ${\alpha}$-factor. $[Orn^6]{\alpha}$-factor 10 demonstrated strong potency with a 7.0-fold increase in halo activity as well as 1.8-fold increase in receptor affinity compared to native ${\alpha}$-factor. For two double substituted analogs, [$Glu^6,{\small{D}}-Ala^9$]${\alpha}$-factor 12 showed the slightly decreased potency in halo activity compared to analog 10, whereas [$Orn^6,{\small{D}}-Ala^9$]${\alpha}$-factor 11 exhibited 15-fold higher halo activity as well as nearly 3-fold higher receptor affinity compared to native ${\alpha}$-factor.

• Journal of agriculture & life science
• /
• v.50 no.2
• /
• pp.151-160
• /
• 2016

Inflammatory cytokines play a major role in osteoclastogenesis, leading to the bone resorption that is frequently associated with osteoporosis. Sulforaphane, isolated from the Broccoli(Brassica oleracea var. italia) florets, inhibits the production of inflamatory cytokine. In the present study, we determined inhibitory effect of sulforaphane on Receptor activator of nuclear factor κB ligand(RANKL)-induced osteoclast formation. Sulforaphane inhibited the expression of osteoclast marker genes, such as tartrate-resistant acid phosphatase(TRAP), cathepsin K, matrix metalloproteinase 9(MMP-9), and calcitonin receptor in RANKL-induced RAW 264.7 macrophage. Also, sluforaphane inhibited the expression of osteoclast protein, such as TRAP, MMP-9, tumor necrosis factor receptor-associated factor 6(TRAF6) and transcription factor nuclease factor of activated T cells(NFAT)c1. Sulforaphane inhibited RANKL-induced activiation of nuclear factor kappaB(NF-kappaB) by suppression RANKL-mediated NF-kappaB transcriptional acitivation. We are confirmed that sulforaphane inhibits not only transcriptional activity of NF-kappaB but also expressions of the osteoclastogenesis factors(TRAP, cathepsin K, MMP-9, calcitonin, TRAF6) and trranscription factor NFATc1.

• Development and Reproduction
• /
• v.3 no.1
• /
• pp.95-100
• /
• 1999

Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor $\beta$ (TGF-$\beta$) superfamily. It has been known that GDF-9 is a growth factor having a crucial role in normal folliculogenesis and its expression is oocyte-specific. The present study was aimed to elucidate the expression of GDF-9 mRNA in the mouse primordial follicles as well as in the other developmental stages. The semiquantitative analysis of GDF-9 mRNA expression was conducted. Total RNA was extracted from the ICR mice ovaries at gestational day 19, postnatal day 1, day 10, day 21, and day 28, and RT-PCR was performed to measure GDF-9 and $\beta$-actin mRNA levels. Level of GDF-9 mRNA were normalized against the level of $\beta$-actin mRNA, and compared among different stages. GDF-9 mRNA was detected in all samples including the fetal ovaries that mainly consists of primordial follicles. The highest level of mRNA was observed in ovaries obtained at day 10 that mainly consists of growing follicles. The present result suggests that GDF-9 may play an important role in the early stage of folliculogenesis.

• BMB Reports
• /
• v.48 no.10
• /
• pp.559-564
• /
• 2015

We investigated the effects of mangiferin on the expression and activity of metalloproteinase (MMP)-9 and the invasion of tumor necrosis factor (TNF)-$\alpha$-stimulated human LNCaP prostate carcinoma cells. Reverse-transcription polymerase chain reaction (RT-PCR) and western blot analysis showed that mangiferin significantly reversed TNF-$\alpha$-induced mRNA and protein expression of MMP-9 expression. Zymography data confirmed that stimulation of cells with TNF-$\alpha$ significantly increased MMP-9 activity. However, mangiferin substantially reduced the TNF-$\alpha$-induced activity of MMP-9. Additionally, a matrigel invasion assay showed that mangiferin significantly reduced TNF-$\alpha$-induced invasion of LNCaP cells. Compared to untreated controls, TNF-$\alpha$-stimulated LNCaP cells showed a significant increase in nuclear factor-${\kappa}B$ (NF-${\kappa}B$) luciferase activity. However, mangiferin treatment markedly decreased TNF-$\alpha$-induced NF-${\kappa}B$ luciferase activity. Furthermore, mangiferin suppressed nuclear translocation of the NF-${\kappa}B$ subunits p65 and p50. Collectively, our results indicate that mangiferin is a potential anti-invasive agent that acts by suppressing NF-${\kappa}B$-mediated MMP-9 expression.

• Clinical and Experimental Reproductive Medicine
• /
• v.12 no.1
• /
• pp.47-70
• /
• 1985

This study was presented of the 1,872 cases of infertile couples who visited and examined at the sterility clinic of Department of Obstetrics & Gynecology, Seoul National University Hospital from Sept., 1980 to Dec., 1983. Age, duration of infertility, past medical history, and other general factors were analyzed, and the factors responsible for infertility were classified and discussed. Mode of treatment, outcome of pregnancy, pregnancy rate responsible for each factor were also presented. The results were as follows: 1) The infertility was primary in 1,128, or 60.3% and secondary in 744, or 39.7%. 2) The age between 26 and 30 years of age comprised about one half of the total patients. 3) The duration of infertility between 1 and 4 years comprised about three quarters of the total patients, and the mean duration was 3.8 years. 4) The most common medical history in primary infertility was tuberculous disease, and that in secondary infertility was history of previous laparotomy. 5) About two thirds of antecedent pregnancies were abortion. 6) The major etiologic factor of infertility were male factor in 12.3%, tubal factor in 38.8%, ovulatory failure in 25.4%, uterine factor in 8.8%, cervical factor in 5.2%, peritoneal factor in 9.5%, and no demonstrable cause in 11.3%. 7) The types of male factor were azoospermia in 61.6%, oligospermia in 25.8%, low motility in 11.6%, and other abnormality in 1.0%. 8) The types of ovulatory failure were ovarian failure in 7.4%, hypothalamo-pituitary failure in 8.1 %, hypothalamo-pituitary dysfunction (including Polycystic ovarian syndrome) in 30.2%, and hyperprolactinemia in 22.4%. 9) The types of uterine factor were endometrial tuberculosis in 27.5%, uterine synechia in 33.8%, uterine anomaly in 19.7%, myoma and polyp in 9.1 %, and luteal phase defect in 9.9%. 10) The types of peritoneal factor were pelvic adhesion in 80.9% and endometriosis in 19.6%. 11) Surgeries were done in 408 patients, and they were salpingolysis, lysis of extraadnexal adhesion, salpingostomy, fimbrioplasty, ovarian wedge resection for polycystic ovarian disease, tubo-tubal anastomosis, and tubo-uterine implantation in orders. 12) 243 pregnancies were achieved during the infertility work-up, of which livebirth was 46.5%, ectopic pregnancy was 7.4%, spontaneous abortion was 7.8%, and on-going pregnancy or lost to follow-up was 36.2%. 13) Pregnancy rates in various factors were male factor in 18.7%, ovulatory factor in 31.7%, tubal factor in 24.2%, uterine factor in 34.6%, cervical factor in 19.0%, peritoneal factor in 29.0%, combined factors in 10.5%, and unexplained infertility in 37.1%. Pregnancy rate in whole patients was 25.2%.