• Journal of Life Science
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• v.28 no.12
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• pp.1416-1423
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• 2018

With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

• Economic and Environmental Geology
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• v.55 no.1
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• pp.63-76
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• 2022

Dacitic tuffs, 97 to 118 m thick, were recovered from the lower part of the subsurface Seongdongri Formation, Janggi Basin, which was drilled to assess the potential for underground storage of carbon dioxide. The tuffs are divided into four depositional units(Unit 1 to 4) based on internal structures and particle componentry. Unit 1 and Units 3/4 are ignimbrites that accumulated in subaerial and subaqueous settings, respectively, whereas Unit 2 is braided-stream deposits that accumulated during a volcanic quiescence, and no dacitic tuff is observed. A series of analysis shows that mordenite and clinoptilolite mainly fill the vesicles of glass shards, suggesting their formation by replacement and dissolution of volcanic glass and precipitation from interstitial water during burial and diagenesis. Glass-replaced clinoptilolite has higher Si/Al ratios and Na contents than the vesicle-filling clinoptilolite in Units 3. However, the composition of clinoptilolite becomes identical in Unit 4, irrespective of the occurrence and location. This suggests that the Si/Al ratio and pH in the interstitial water increased with time because of the replacement and leaching of volcanic glass, and that the composition of interstitial water was different between the eastern and western parts of the basin during the formation of the clinoptilolite in Units 1 and 3. It is also inferred that the formation of the two zeolite minerals was sequential according to the depositional units, i.e., the clinoptilolite formed after the growth of mordenite. To summarize, during a volcanic quiescence after the deposition of Unit 1, pH was higher in the western part of the basin because of eastward tilting of the basin floor, and the zeolite ceased to grow because of the closure of the pore space as a result of the growth of smectite. On the other hand, clinoptilolite could grow in the eastern part of the basin in an open system affected by groundwater, where braided stream was developed. Afterwards, Units 3 and 4 were submerged under water because of the basin subsidence, and the alkali content of the interstitial water increased gradually, eventually becoming identical in the eastern and western parts of the basin. This study thus shows that volcanic deposits of similar composition can have variable distribution of zeolite mineral depending on the drainage and depositional environment of basins.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.9
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• pp.1237-1247
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• 2012

The objective of this study was to evaluate the predictions of dry matter intake (DMI) and average daily gain (ADG) of Vietnamese Yellow (Vang) purebred and crossbred (Vang with Red Sindhi or Brahman) bulls fed under Vietnamese conditions using two levels of solution (1 and 2) of the large ruminant nutrition system (LRNS) model. Animal information and feed chemical characterization were obtained from five studies. The initial mean body weight (BW) of the animals was 186, with standard deviation ${\pm}33.2$ kg. Animals were fed ad libitum commonly available feedstuffs, including cassava powder, corn grain, Napier grass, rice straw and bran, and minerals and vitamins, for 50 to 80 d. Adequacy of the predictions was assessed with the Model Evaluation System using the root of mean square error of prediction (RMSEP), accuracy (Cb), coefficient of determination ($r^2$), and mean bias (MB). When all treatment means were used, both levels of solution predicted DMI similarly with low precision ($r^2$ of 0.389 and 0.45 for level 1 and 2, respectively) and medium accuracy (Cb of 0.827 and 0.859, respectively). The LRNS clearly over-predicted the intake of one study. When this study was removed from the comparison, the precision and accuracy considerably increased for the level 1 solution. Metabolisable protein was limiting ADG for more than 68% of the treatment averages. Both levels differed regarding precision and accuracy. While level 1 solution had the least MB compared with level 2 (0.058 and 0.159 kg/d, respectively), the precision was greater for level 2 than level 1 (0.89 and 0.70, respectively). The accuracy (Cb) was similar between level 1 and level 2 (p = 0.8997; 0.977 and 0.871, respectively). The RMSEP indicated that both levels were on average under-or over-predicted by about 190 g/d, suggesting that even though the accuracy (Cb) was greater for level 1 compared to level 2, both levels are likely to wrongly predict ADG by the same amount. Our analyses indicated that the level 1 solution can predict DMI reasonably well for this type of animal, but it was not entirely clear if animals consumed at their voluntary intake and/or if the roughness of the diet decreased DMI. A deficit of ruminally-undegradable protein and/or a lack of microbial protein may have limited the performance of these animals. Based on these evaluations, the LRNS level 1 solution may be an alternative to predict animal performance when, under specific circumstances, the fractional degradation rates of the carbohydrate and protein fractions are not known.

• Journal of Environmental Health Sciences
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• v.28 no.1
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• pp.51-65
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• 2002

This study was carried out to apply some basic physical and chemical treatment options including Fenton's oxidation, and to evaluate the performances and the characteristics of organic and nitrogen removal using lab-scale biological treatment system such as complete-mixing activated sludge and sequencing batch reactor(SBR) processes for the treatment of leachate from a municipal waste landfill in Gyeongnam province. The results were as follows: Chemical coagulation experiments using aluminium sulfate, ferrous sulfate and ferric chloride resulted in leachate CO $D_{Cr}$ removal of 32%, 23% and 21 % with optimum reaction dose ranges of 10,000～15,000 mg/$\ell$, 1,000 mg/$\ell$ and 500～2,000 mg/$\ell$, respectively. Fenton's oxidation required the optimum conditions including pH 3.5, 6 hours of reaction time, and hydrogen peroxide and ferrous sulfate concentrations of 2,000 ～ 3,000 mg/$\ell$ each with 1:1 weight ratio to remove more than 50% of COD in the leachate containing CO $D_{Cr}$ between 2,000 ～ 3,000 mg/$\ell$. Air-stripping achieved to remove more than 97% of N $H_3$-N in the leachate in spite of requiring high cost of chemicals and extensive stripping time, and, however, zeolite treatment removing 94% of N $H_3$-N showed high selectivity to N $H^{+}$ ion and much faster removal rate than air-stripping. The result from lab-scale experiment using a complete-mixing activated sludge process showed that biological treatability tended to increase more or less as HRT increased or F/M ratio decreased, and, however, COD removal efficiency was very poor by showing only 36% at HRT of 29 days. While COD removal was achieved more during Fenton's oxidation as compared to alum treatment for the landfill leachate, the ratio of BOD/COD after Fenton's oxidation considerably increased, and the consecutive activated sludge process significantly reduced organic strength to remove 50% of CO $D_{Cr}$ and 95% of BO $D_{5}$ . The SBR process was generally more capable of removing organics and nitrogen in the leachate than complete-mixing activated sludge process to achieve 74% removal of influent CO $D_{Cr}$ , 98% of BO $D_{5}$ and especially 99% of N $H_3$-N. However, organic removal rates of the SBR processes pre-treated with air-stripping and with zeolite were not much different with those without pre-treatment, and the SBR process treated with powdered activated carbon showed a little higher rate of CO $D_{Cr}$ removal than the process without any treatment. In conclusion, the biological treatment process using SBR proved to be the most applicable for the treatment of organic contents and nitrogen simultaneously and effectively in the landfill leachate.e.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.220-220
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• 2013

Surface-normal transmission electro-absorption modulator (EAM) are attractive for high-definition (HD) three-dimensional (3D) imaging application due to its features such as small system volume and simple epitaxial structure [1,2]. However, EAM in order to be used for HD 3D imaging system requires uniform modulation performance over large area. To achieve highly uniform modulation performance of EAM at the operating wavelength of 850 nm, it is extremely important to remove the GaAs substrate over large area since GaAs material has high absorption coefficient below 870 nm which corresponds to band-edge energy of GaAs (1.424 eV). In this study, we propose and experimentally demonstrate a transmission EAM in which highly selective backside etching methods which include lapping, dry etching and wet etching is carried out to remove the GaAs substrate for achieving highly uniform modulation performance. First, lapping process on GaAs substrate was carried out for different lapping speeds (5 rpm, 7 rpm, 10 rpm) and the thickness was measured over different areas of surface. For a lapping speed of 5 rpm, a highly uniform surface over a large area ($2{\times}1\;mm^2$) was obtained. Second, optimization of inductive coupled plasma-reactive ion etching (ICP-RIE) was carried out to achieve anisotropy and high etch rate. The dry etching carried out using a gas mixture of SiCl4 and Ar, each having a flow rate of 10 sccm and 40 sccm, respectively with an RF power of 50 W, ICP power of 400 W and chamber pressure of 2 mTorr was the optimum etching condition. Last, the rest of GaAs substrate was successfully removed by highly selective backside wet etching with pH adjusted solution of citric acid and hydrogen peroxide. Citric acid/hydrogen peroxide etching solution having a volume ratio of 5:1 was the best etching condition which provides not only high selectivity of 235:1 between GaAs and AlAs but also good etching profile [3]. The fabricated transmission EAM array have an amplitude modulation of more than 50% at the bias voltage of -9 V and maintains high uniformity of >90% over large area ($2{\times}1\;mm^2$). These results show that the fabricated transmission EAM with substrate removed is an excellent candidate to be used as an optical shutter for HD 3D imaging application.

• Korean Journal of Ecology and Environment
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• v.43 no.1
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• pp.82-90
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• 2010

The ecological characteristics of the Korean Aucha perch, Coreoperca herzi, were determined in order to estimate stock of the mid-upper system of the Seomjin River. The age was determined by counting the otolith annuli. The oldest fish observed in this study was 5 years old. Relationships between body length (BL) and body weight (BW) were $BW=0.0195BL^{3.08}$ ($R^2=0.966$) (p<0.01). Relationships between the otolith radius (R) and body length (BL) were BL=3.882R+1.66 ($R^2=0.944$). The von Bertalanffy growth parameters estimated from a non-linear regression method were $L_{\infty}=19.68\;cm$, $W_{\infty}=188.64\;g$, $K=0.17\;year^{-1}$ and $t_0=-1.46$ year. Therefore, growth in length of the fish was expressed by the von Bertalanffy's growth equation as $L_t=19.68$ ($1-e^{-0.17(t+1.46)}$) ($R^2=0.997$). The annual survival rate (S) was estimated to be $0.666\;year^{-1}$. The instantaneous coefficient of natural mortality (M) of estimated from the Zhang and Megrey method was $0.346\;year^{-1}$, and instantaneous coefficient of fishing mortality (F) was calculated $0.061\;year^{-1}$. From the estimates of survival rate (S), the instantaneous coefficient of total mortality(Z) was estimated to be $0.407\;year^{-1}$.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.448-468
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• 1996

To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

• Journal of Korean Society of Forest Science
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• v.83 no.3
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• pp.286-298
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• 1994

The object of this study was to examine the user's impacts on the environmental deteriorations of trail at Ticket Office - Paekryunsa (Temple) Hyangch$\hat{o}$kpong - Dongyupryung - Chilyun Fall area in T$\hat{o}$kyusan National Park. Four trails were sampled in the study area according to the amount of users. Then the user's impacts on trail were measured at each trail. The Ticket Office-Paekryunsa trail was the most used district and followed at Paekryunsa-Hyangch$\hat{o}$kpong trail, Hyangch$\hat{o}$kpong-Dongyupryung trail in descending order. Dongyupryung-Chilyun Fall trail is not used by people because of rest rotation system. The entire width of trail was greater at the more heavily used trail. Maximum depth, cross-sectional area loss, and surface texture and roughness of trail were the highest at Paekryunsa-Hyangch$\hat{o}$kpong trail. Soil hardness, soil acidity, soil moisture content, organic matter content, and exchange canon were influenced by trampling. Soil hardness, soil acidity and exchange canon increased in tramples soil, but content of soil moisture and organic matter decreased therein. Environmental deteriorations of trail were significantly influenced by the amount of users and the slope of trail. Bared lands about $2.000m^2$ were appeared by trampling and camping around Hyangch$\hat{o}$kpong. Effects of carrying of rest rotation system for National Park were partly recognized at Dongyupryung-Chilyun Fall trail.

• The Journal of Korean Academy of Prosthodontics
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• v.51 no.4
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• pp.315-322
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• 2013

Purpose: To evaluate the axial displacement of implant-abutment assembly after cyclic loading in internal tapered connection system. Materials and methods: External butt-joint connection implant and internal tapered connection implant were connected with three types of abutment for cement-retained prostheses, i.e. external type abutment (Ext group), internal tapered 1-piece abutment (Int-1 group), and internal tapered 2-piece abutment (Int-2 group). For each group, 7 implants and abutments were used. The implantabutments assemblies were clamped into the implant holder for vertical loads. A dynamic cyclic loading was applied for $150{\pm}10N$ at a frequency of 4 Hz. The amount of axial displacement of the abutment into the implant was calculated at each cycle of 0, 5, 10, 50, 100, 1,000, 5,000, and 10,000. A repeated measures analysis of variance (ANOVA) for the overall effect of cyclic loading and the pattern analysis by linear mixed model were used for statistical analysis. Differences at P<.05 were considered statistically significant. Results: The mean axial displacement after 10,000 cycles were $0.714{\pm}0.488{\mu}m$ in Ext group, $5.286{\pm}1.604{\mu}m$ in Int-1 group, and $11.429{\pm}1.902{\mu}m$ in Int-2 group. In the pattern analysis, Int-1 and Int-2 group showed continuous axial displacement at 10,000 cycles. There was no declining pattern of axial displacement in the Ext group. Conclusion: The pattern of linear mixed model in Ext group showed no axial displacement. There were continuous axial displacements in abutment-implant assemblies in the Int-1 and Int-2 group at 10,000 cycles. More axial displacement was found in Int-2 group than in Int-1 group.

• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.