• Korean Journal of Animal Reproduction
• /
• v.27 no.4
• /
• pp.299-307
• /
• 2003

The effects of additions/deletions in glycosylated residues of recombinant human EPO (rhEPO) produced in CHO-K1 on their secretion were examined. hEPO cDNA was amplified from human liver mRNA and cloned into the pCR2.1 TOPO. Using overlapping-extension site-directed mutagenesis method, glycosylation sites at 24th, 38th, 83rd, and 126th were respectively or accumulatively removed by substituting its asparagine (or serine) with glutamine. To add novel glycosylation sites, 69 and 105th leucine was mutated to asparagine. Mutant and wild type rhEPO constructs were cloned into the pcDNA3 expression vector with CMV promoter and transfected into CHO cell line, CHO-K1, to produce mutant rhEPO mutant rhEPO proteins. Enzyme-linked immunosorbant assay (ELISA) and Western analysis with monoclonal anti-EPO antibody were performed using supernatants of the cultures showing transient and stable expressions respectively. Addition of novel glycosylation reduced rhEPO secretion dramatically while deletion mutants had little effect except some double deletion mutants ($\Delta$24/83 and $\Delta$38/83) and triple mutant ($\Delta$24/38/83). This fact suggests that not single but combination of changes in glycosyl groups affect secretion of rhEPO in cell culture, possibly via changes in their conformations.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.6-6
• /
• 2001

Oocyte freezing has become a prevalent source for related reproductive technologies. This study was carried out to evaluate viability of post-thawed bovine oocyte injected DTT-treated sperm following by two different activation stimuli (Group 1, 5 M ionomycin, 5 min ＋ CR1aa, 3 h . 1.9 mM dimetylaminopurine (DMP), 3 h; group 2, ionomycin ＋ 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CHX), 5h). The techniques of ultra-rapid freezing used in this study were essentially similar to those of described by Vajta et al (Theriogenology 1999; 52:939-948), Denuded oocytes at 22 h of culture were exposed to cryoprotectant (3.2 M Ethylene glycol, 2.36 M DMSO, 0.6 M sucrose), and followed by freezing in electron microscopic grid. After thawing the oocytes were transferred back into the drop of maturation medium and cultured for additional 2 h before being subjected to ICSI. All eggs were then cultured in CRlaa medium, and transferred into M199+10% FCS on day 4. The culture was maintained until day 9. In Experiment 1, frozen-ICSI eggs were compared on development into blastocyst to those of unfrozen and IVF control. Those eggs were activated with the method of group 2. A higher proportion of unfrozen-ICSI and IVF eggs developed into cleavage and blastocysts than of frozen-ICSI eggs (65% and 13%; 71% and 23% vs. 39% and 8%; P<0.05). In Experiment 2, development and ploidy of embryos made from group 1 were compared to those from group 2. Between groups there did not differ on the rates of development, however, chromosomal abnormality in group 1 was significantly higher than in group 2 (49% vs. 30%; P<0.05). The present result suggests that frozen bovine oocytes can be used for ICSI.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.87-87
• /
• 2001

도살장에서 구입한 소의 난소로부터 회수한 미성숙 난포란을 체외에서 성숙, 수정시킨 후, 2~8세포기 수정란을 CR$_1$aa 체외배양액에 일정량의 aesculetin, taurine을 첨가하여 체외배양을 실시한 두 처리구간의 항산화 효과를 비교 검토하고, aesculetin과 taurine에 growth factor(EGF, PDGF)를 첨가배양하여 체외수정란의 체외발육에 항산화제와 growth factor의 상호작용 효과를 검토하였다. 대조구, aesculetin(1$\mu\textrm{g}$/$m\ell$) 및 taurine(2.5mM)을 첨가하여 체외수정란을 체외배양시킨 결과 상실배이상 발육된 체외발육율은 각각 46.5%, 64.3% 및 60.5%로서 대조구보다 aesculetin과 taurine 첨가구가 유의하에 높은 성적을 얻어, aesculetin과 taurine이 체외수정란의 체외발육에 높은 효과를 나타냈다. 대조구, aesculetin (1$\mu\textrm{g}$/$m\ell$) ＋ PDGF (1ng/$m\ell$) 첨가구, aesculetin (l$\mu\textrm{g}$/$m\ell$) ＋ EGF (10ng/$m\ell$)첨가구, taurine (2.5mM) ＋PDGF (1ng/$m\ell$) 첨가구 및 taurine (2.5mM)＋EGF (10ng/$m\ell$) 첨가구에서 배반포기 발육율은 각각 46.0%, 70.0%, 58.0%, 56.0 및 66.0%로써 처리구가 무첨가구에 비해 유의하게 높은 성적을 얻어(P<0.05), 항산화제와 growth factor의 첨가 배양이 체외수정란의 체외발육에 상승효과가 있음이 입증되었다. 배양액에 천연추출 aesculetin과 상품화된 aesculetin을 첨가하여 체외배양을 실시한 결과 배반포기 발육율은 대조구, 천연추출 aesculetin 및 상품화된 aesculetin 첨가구에서 각각 38.6%, 54.6% 및 53.5%로써 첨가구가 무첨가구보다 높은 성적을 얻어, aesculetin의 항산화 효과가 입증되었다(P<0.05). 그러나 모든 처리구에서 배반포까지 발육된 체외수정란의 세포수에는 커다란 차이가 인정되지 않았다.

• The Korean Journal of Physiology and Pharmacology
• /
• v.9 no.2
• /
• pp.131-135
• /
• 2005

Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in $Ca^{2+}$ homeostasis and/or $Ca^{2+}-activated$ kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific $K^+$ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance $Ca^{2+}$-activated $K^+$ ($BK_{Ca}$) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance $K^{+}$ channels (106 pS of chord conductance at +40 mV in physiological $K^+$ gradient). The 106 pS channels were activated by membrane potential and $[Ca^{2+}]_i$, consistent with the known properties of $BK_{Ca}$ channels. $BK_{Ca}$ channels in CRL-1474 were positively regulated by adenylate cyclase activator ($10{\mu}M$ forskolin), 8-Br-cyclic AMP ($300{\mu}M$) or 8-Br-cyclic GMP ($300{\mu}M$). These results suggest that human skin fibroblasts (CR-1474) have typical $BK_{Ca}$ channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.6
• /
• pp.2467-2471
• /
• 2012

Introduction: Although bleomycin/etoposide/cisplatinum (BEP) chemotherapy is established as the standard treatment for germ cell tumours, it requires significant experience in administration and toxicity management to maintain optimal dose intensity. A retrospective review of 30 patients was conducted at UKMMC to study treatment outcomes. Methods & Materials: Patients with GCTs and treated with at least two cycles of BEP chemotherapy between January 2003 and Oct 2009 were eligible for this study. Patients received 4-6 cycles of bleomycin 30,000IU IV D1, D8 & D15 and either etoposide $100mg/m^2$ IV D1-D5 and cisplatin $20mg/m^2$ IV D1-D5 (5 day BEP regimen) or etoposide $165mg/m^2$ D1-D3 and cisplatin $50mg/m^2$ D1-3 (3 day BEP regimen) every three weeks per cycle. All patients received prophylactic granulocyte colony-stimulating factor (GCSF) from days 6 to 10 of each cycle. The overall response rates, 2 year progression-free survival and overall survival of the whole cohort were assessed. Results: Thirty patients fulfilled the inclusion criteria. Non-seminomatous GCTs comprised 93.3% of cases and gonadal and mediastinal primary sites were the most common. Sixty percent were classified as IGCCCG poor risk disease. Median follow-up was 26.6 months. The overall response rate (CR+PR) was 70%. The two year PFS and OS were 70% and 66%. There was a significant difference in terms of the overall response rate (85% vs 40%, p = 0.03) and in PFS (94.7% vs 50%, p = 0.003) between gonadal and extragonadal primary sites. Conclusion: It is possible to achieve outcomes similar to those in international clinical trials with close monitoring and good supportive care of patients undergoing BEP chemotherapy. There is a strong argument for patients with IGCCCG poor prognosis disease to be treated in specialist tertiary centres to optimize treatment outcomes.

• Journal of Powder Materials
• /
• v.23 no.3
• /
• pp.240-246
• /
• 2016

In order to improve the high-temperature oxidation stability, sintered 434L stainless steel is studied, focusing on the effect of the addition of metallic oxides to form stable oxide films on the inner particle surface. The green compacts of Fecralloy powder or amorphous silica are added on STS434L and oxidized at $950^{\circ}C$ up to 210 h. The weight change ratio of 434L with amorphous silica is higher than that of 434L mixed with Fecralloy, and the weight increase follows a parabolic law, which implies that the oxide film grows according to oxide diffusion through the densely formed oxide film. In the case of 434L mixed with Fecralloy, the elements in the matrix diffuse through the grain boundaries and form $Al_2O_3$ and Fe-Cr oxides. Stable high temperature corrosion resistance and electrical resistivity are obtained for STS434L mixed with Fecralloy.

• Ocean and Polar Research
• /
• v.25 no.1
• /
• pp.21-30
• /
• 2003

Seasonal fluctuations of physico-chemical and biological aspects of the environment were studied in Vladivostok harbour (Golden Horn Bay, the East Sea/Sea of Japan). The benthic community structure was described with a focus on size-spectra (bacteria, meio- and macrofauna) related with the chemical environment and chemical fluxes in sediment and to reveal their possible ecological role in the process of bioremediation of the environment. Samples from two sites with different concentrations of heavy metals (Fe, Zn, Cu, Pb, Mn, Cr, Ni Cd, Co) and petroleum hydrocarbon were assessed by a number of methods. These included plate counts of culturable bacteria, observation through a scanning electron (SEM) and transmission electron microscope (TEM). These approaches were complemented with microscopic assessments of the diversity of the benthic community. The specific communities had a limited number of species, tolerant to abnormally high levels of toxic compounds. The dominant species were presented by several sho.1-lived small polychaetes (Capitella capitata) and nematodes (Oncholaimium ramosum). The highest population density was recorded in microbenthos, in various diatoms, various physiological groups of bacteria which participate in biomineralization: marine heterotrophic bacteria, which oxidized oil, black oil in addition to groups resistant to heavy metals. They have the entire set of mechanisms for neutralizing the negative effect of those compounds, forming the detrital food web and biogeochemical circulation of material in sediments, which results in the biological self-recycling of sea basins. Macro- and meiobenthic organisms were more sensitive to a greater extent of $H_2S$ and petroleum hydrocarbons than to metal content, but the within-site rankings were the same as those achieved for microbiological analyses.

• Journal of Korean Society of Environmental Engineers
• /
• v.22 no.10
• /
• pp.1799-1808
• /
• 2000

To investigate the applicability of loess as a coagulant. the optimum conditions of coagulation and efficiency comparison of several coagulants (PAC, PACS, LAS) in chemical treatment of livestock wastewater were performed. As a result, the optimum mixing ratio of loess and lime(CaO) was 3:7. The optimum dosage of the mixed coagulant was 30 g/L. The optimum speed for rapid mixing of the mixed coagulant was 200 rpm at 1 min. and the optimum speed for slow mixing was 50 rpm at 10 min. The mixed coagulant showed the removal efficiency of turbidity, SS, BOD, $COD_{cr}$. T-P and TKN to 95.8%, 92.5%, 71.6%, 71.1%, 98.2% and 32.5%, respectively, which was better than other several coagulants. The mixed coagulant was possible to use as substitutional coagulants of traditional coagulant, and the producted sludge can use as a soil amendments.

• Proceedings of the Korean Environmental Sciences Society Conference
• /
• 2007.05a
• /
• pp.375-379
• /
• 2007

폐비닐 여재(Synthetic Waste Polyethylene Media)를 적용한 부착성장식 공정의 유기물 및 질소${\cdot}$인 제거특성에 관한 Pilot Plant 실험 결과 다음과 같은 결론을 얻을 수 있었다. 1) 제거효율은 $RUNI1{\sim}3$에서 $COD_{cr}$ 91.4, 92.4, 93.9%, T-N 56.9, 61.4, 65.1%, T-P는 모든 단계에서 약 45%이상 제거되어 부하변동시에 강한 대처능력을 나타내었다. 2) BOD용적부하 $0.18{\sim}0.40kg/m^3{\cdot}d$, COD용적부하 $0.28{\sim}0.53kg/m^3{\cdot}d$, ${NH_4}^{+}-N$용적부하 $0.12kg/m^3{\cdot}d$을 나타내었다. 3) 도시하수 처리를 위한 생물학적 질소 및 인 제거공법을 본 여재를 이용하여 공정설계시 고려할 사항들과 적절한 대처방법을 다음과 같이 제시할 수 있다. - 1차 침전지에서 유기질소 및 유기인 농도가 높다면 유기질소 및 인 부하량을 감소시키기 위하여 1차 침전지를 설치하되 질소 및 인제거에 유리한 $BOD_5/T-N$, $BOD_5/T-P$를 유지할 수 있도록 체류시간을 1시간미만으로 설계하는 것이 경제적이라고 판단된다. 만일 유기질소와 유기인의 함량이 낮다면 1차 침전지는 제외하는 것이 유리할 것이다. - 여재의 배치는 폭기조에서 용존산소의 균일분포와 슬러지의 적정탈리를 위해 여재를 상하로 배치하거나 또는 여재끼리 일정 간격을 두어 배치하는 것이 바람직하다. 농촌에서의 처분이 문제시 되고 있는 폐비닐을 적용한 본 연구에서의 수처리특성은 기존 하수처리공정에서의 제거효율에 상응하는 처리특성을 나타내었다. 또한 폐비닐 처분의 문제를 해결할 수 있을뿐만 아니라 하수처리시에도 부하변동 등에 강한 대처능력을 나타내어 기존의 하수처리공정에 대체가능성을 나타내었다.

• Journal of Environmental Science International
• /
• v.25 no.11
• /
• pp.1563-1573
• /
• 2016

The aim of this study was to evaluate the occurrence of vanadium in Jeju Island groundwater, focusing on the spatio-temporal patterns and geochemical controlling factors of vanadium. For this, we collected two sets of groundwater data: 1) concentrations of major constituents of 2,595 groundwater samples between 2008 and 2014 and 2) 258 groundwater samples between December 2006 and June 2008. The concentrations of groundwater vanadium were in the range of $0.2{\sim}71.0{\mu}g/L$ (average, $12.0{\mu}g/L$) and showed local enrichments without temporal/seasonal variation. This indicated that vanadium distribution was controlled by 1) the geochemical/mineralogical composition and dissolution processes of original materials (i.e., volcanic rock) and 2) the flow and chemical properties of groundwater. Vanadium concentration was significantly positively correlated with that of major ions ($Cl^-$, $Na^+$, and $K^+$) and trace metals (As, Cr, and Al), and with pH, but was negatively correlated with $NO_3-N$ concentration. The high concentrations of vanadium (>$15{\mu}g/L$) occurred in typically alkaline groundwater with high pH (${\geq}8.0$), indicating that a higher degree of water-rock interaction resulted in vanadium enrichment. Thus, higher concentrations of vanadium occurred in groundwater of $Na-Ca-HCO_3$, $Na-Mg-HCO_3$ and $Na-HCO_3$ types and were remarkably lower in groundwater of $Na-Ca-NO_3$(Cl) type that represented the influences from anthropogenic pollution.