• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.29-36
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• 2011

Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections on urinary track, cornea, respiratory track, and burn wound site, and mainly relies on quorum sensing (QS) for its virulence. To control the infectivity of P. aeruginosa, we previously synthesized the structural analogues of a major QS signal, N-3-oxododecanoyl homoserine lactone (3OC12-HSL) to use as a QS inhibitor. Two of them (5b and 5f) had been confirmed to have an inhibitory effect on LasR, a major QS signal receptor of P. aeruginosa in the screening by the recombinant Escherichia coli reporter. To further evaluate these compounds, we tested their efficacy to control the QS and virulence of P. aeruginosa. Unlike the result from E. coli reporter, both 5b and 5f failed to affect the LasR activity in P. aeruginosa, but instead they selectively affected the activity of QscR, another 3OC12-HSL receptor of P. aeruginosa. Interestingly, their effect on QscR was complex and opposite to what we obtained with E. coli system. Both 5b and 5f enhanced the QscR activity at the low concentration range (< 10 ${\mu}m$), but high concentration of 5f (${\approx}$1 mM) strongly inhibited QscR. While 5b and 5f didn't affect the production of proteases, the key virulence factor, they significantly reduced the biofilm formation that is important in mediating chronic infections. Especially, 5f inhibited the initial attachment of P. aeruginosa, rather than the biofilm maturation. Based on our results, we suggest that 5f can be applied for an anti-biofilm agent without increasing virulence of P. aeruginosa.

• Journal of Life Science
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• v.26 no.11
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• pp.1259-1268
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• 2016

In this study, we investigated antimicrobial peptide from the acidified muscle extract of Mytilus coruscus, which mostly inhabits China, Japan, and Korea, to develop a natural product-derived antibiotics substitution in terms of its abuse and restriction. Antimicrobial peptide was purified by $C_{18}$ reversed-phase high-performance liquid chromatography and was detected as having a molecular mass of 6,701 Da by MALDI-TOF/MS. The N-terminal amino acid sequence of the purified peak was obtained from edman degradation, and 20 identified residues shown 100% identity with the N-terminus region of sperm-specific protein and protamine-like PL-II/PL-IV precursor of Mytilus californianus. We also identified 60 open-reading frame (ORF) encoding amino acids with 183 bp of purified peptide based on the obtained amino acid residues. The amino acid sequence of ORF showed 100% and the nucleotide sequence revealed 97.2% identity with the protamine-like PL-II/PL-IV precursor of Mytilus californianus. Synthesized antimicrobial peptide showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus (minimal effective concentration [MEC], $20.8{\mu}g/ml$), Bacillus subtilis (MEC, $0.2{\mu}g/ml$), Streptococcus mutans (MEC, $0.2{\mu}g/ml$), gram-negative bacteria including Pseudomonas aeruginosa (MEC, $5.7{\mu}g/ml$), Escherichia coli (MEC, $2.6{\mu}g/ml$) and fungi, Candida albicans (MEC, $56.3{\mu}g/ml$). In addition, synthesized peptide showed stable activities under heat and salt conditions against gram-positive and gram-negative bacteria, but was inhibited by salt against only C. albicans. With these results, isolated peptide from M. coruscus could be an alternative agent to antibiotics for defending against pathogenic microorganisms, and helpful information to understand the innate immune system of marine invertebrates.

• Korean Journal of Poultry Science
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• v.37 no.3
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• pp.255-263
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• 2010

Inclusion body hepatitis-hydropericardium syndrome (IBH-HPS) is an acute viral disease usually found in broilers aged from 3 to 5 weeks and causes up to 75% mortality. Among the 12 serotypes of fowl adenovirus group 1, serotype-4 (FAdV-4) was identified as a primary agent of IBH-HPS and was usually isolated in IBH-HPS cases in Korea since 2007. To prevent these IBH-HPS outbreaks in Korea, we developed the FAdV-4 inactivated vaccine using Korean isolate (ADL070244) and evaluated the efficacy of this vaccine. For the efficacy test, 2-week-old specific-pathogen-free (SPF) chickens intramuscularly inoculated with 1 or 2 dose of inactivated vaccine were used and challenged with FAdV-4 through either intramuscular or oral route at 2 weeks after vaccination. The vaccine induced good seroconversion which was confirmed by agar gel precipitation (AGP) test. In addition, the vaccine could decrease the FAdV-4 detection rate and histological lesion severity such as lymphocyte infiltration and necrosis in the liver comparing with those of non-vaccination group. Based on the current results, the developed FAdV-4 inactivated vaccine in this study was effective in the terms of reduction of virus detection rate and histological lesions severity. However, it was difficult to confirm the efficacy of the vaccine clearly because of no mortality and clinical signs in the non-vaccinated group after challenge. Therefore, we need further study to develop a standard challenged model system which could clearly evaluate the efficacy of the vaccines for FAdV-4.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.21-26
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• 2010

As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

• Korean Journal of Poultry Science
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• v.35 no.1
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• pp.85-99
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• 2008

Avian reovirus (ARV) is a causative agent of viral arthritis/tenosynovitis, and malabsorption syndrome in broiler. The characteristics of malabsorption syndrome caused by ARV are diarrhea, poor feed conversion and stunting. Therefore, ARV infection has been recognized as one of the most important disease in the poultry industry because of economical losses. However, few study of ARV infection in broiler industry has been conducted in Korea. To evaluate the presence of ARV infection in broiler farms, epidemiological survey such as serological test and virus isolation has been conducted. For the serological survey using ELISA method, we selected five broiler farms which were located at different area and had a history of growth retardation, lameness, diarrhea and poor feathering. From these farms serum samples were collected at 1 day, 14 days and market age. All these farms had no history of vaccination against ARV. In addition to serological survey, we tried to isolate ARV from birds of designated farms at market age and collected feces and tissue samples such as cecal tonsil, intestine and liver. We were identified ARV by RT-PCR and transmissible electron microscopy. The samples were inoculated into 9-day-old embryonated eggs via the chorioallantoic membrane to observe the pock formation. For the pathogenicity test of ARV isolates, we inoculated with the isolates to the right footpad of 3-week-old SPF chicks and observed clinical signs and pathological changes for 14 days after challenge. Most broilers sampled for serological survey have maternal antibodies which were widely distributed at 1 day and decreased by 14 days. However, at the market age several broiler farms showed fairly high antibody titer against ARV. This increase of antibody titer at market age means the possible infection of ARV during the grow-out period. Among total 15 samples for the isolation of ARV. 2 samples were positive by RT-PCR and finally identified as a ARV. We inoculated these isolates in the SPF birds and observed that the antibody titer was increased from 7 days after challenge. However, we did not find any clinical signs both control and challenge groups. Based on the above results, it is clear that the ARV infection has been circulated in the broiler industry and caused significant economic losses. Further study is needed to evaluate the virulence of the isolates in the digestive system of broiler and the molecular characteristics of isolates.

• Korean Journal of Ecology and Environment
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• v.41 no.3
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• pp.402-411
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• 2008

Grazing effects of two freshwater shellfish, Unio douglasiae (UNIO) and Cipangoplaudina chinese malleata (CCM), on the hibernal diatom communities in eutrophic water was examined in a laboratory. Two animals having different feeding types used in the study were collected from Keum River (Boryeong, Korea), acclimatized in the artificial management system in a laboratory over at least one month, and starved in a laboratory for 2 days before the experiment. Experimental waters, which dominated by Synedra ulna-Scenedesmus sp. (ca. 80%), was collected from eutrophic lake, Lake Ilgam (Seoul) in Feb., 19, 2008 at AM 10:00, and used in the study after the filtration with 1 mm Nylon mesh. Feeding experiments were largely divided into three kinds of animal treatments; five densities of UNIO (U0, U1, U2, U3, U4) and CCM (C0, C1, C2, C3, C4), and four combined densities of two shellfish (M0, M1, M2, M3). U0 and C0 were control (no addition of UNIO or CCM), U1 or C1 (each animal density at 0.5 ind. $L^{-1}$), U2 or C2 (1.0 ind. $L^{-1}$), U3 or C3 (1.5 ind. $L^{-1}$), and U4 or C4 (2.0 ind. $L^{-1}$), respectively. Four combined treatments were consisted of M0 (no animals), M1 (U1+C3), M2 (U2+C2), and M3 (U4+C1), respectively. Under the presence of animal, the concentration of Chl-a and algal abundance were clearly decreased with the increase of UNIO density and the treated time, while in combined group a strong decrease of algal density showed with the increase of UNIO density. Total phytoplankton density shifted as the similar patterns to that of Chl-a concentration (r=0.6527, p<0.0001), however, there showed the differences following a species. There were strong decreases of dominant species Synedra ulna, Scenedesmus sp., Ankistrodesmus falcatus in UNIO treatment group, Diatoma vulgare in combined group, while Cryptomonas ovata in high density of CCM increased about 20% in algal density. Grazing rates (GRs) based on the concentration of Chl-a was depend on the kind of shellfish and treatment time; a strong feeding of CCM showed in the initial stage, and four hours later, UNIO and combined group with high UNIO density showed the high GRs. Interestingly, faeces production of shellfish was highest in combined group with high density of CCM, while their size over 60 ${\mu}m$ was much higher in production magnitude than that less 60 ${\mu}m$. Collectively, these results suggest that two domestic shellfish and its combined treatment have a strong potential as an effective biological controller of diatom bloom in cold eutrophic waters.

• 한국환경농학회:학술대회논문집
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• 2009.07a
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• pp.273-282
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• 2009

The transmissible spongiform encephalopathies (TSEs) disease group are fatal neurodegenerative disorders affecting a wide range of hosts. The group includes kuru and Creutzfeldt-Jakob disease (CJD) in humans, scrapie in sheep and goats and Bovine spongiform encephalopathy (BSE) in cattle. The exact nature of the infectious agent involved in the transmission of these diseases remains controversial. However, a central event in their pathogenesis is the accumulation in infected tissues of an abnormal form of a host-encoded protein, the prion protein (PrP). Whereas the normal cellular protein is fully sensitive to protease ($PrP^{sen}$), the disease-associated prion protein ($PrP^d$) is only partly degraded ($PrP^{res}$), its amino-terminal end being removed. BSE was first reported in the mid-80s in the UK. Ten years later, a new form of human prion disease, variant CJD (vCJD) developed in the wake of the BSE epidemic, and there is now strong scientific evidence that vCJD was initiated by the exposure of humans to BSE-infected tissues, thus indicating a zoonotic disease. However, the ban on the feeding of animal-derived proteins to ruminants, and the apparent lack of vertical transmission of BSE, have led to a decline in the incidence of the disease within cattle herd and therefore, an assumed decreased risk for human contacting vCJD. The origin of the original case(s) of BSE still remains an enigma even though three hypotheses have been raised. Hypotheses are i) sheep- or goat-derived scrapie-infected tissues included in meat and bone meal fed to cattle, ii) a previously undetected sporadic or genetic bovine TSE contaminating cattle feed or iii) originating from a human TSE through animal feed contaminated with human remains. A host cellular membrane protein ($PrP^C$), which is abundant in central nervous system tissue, appear to be conformationally altered in the diseased host into a prion protein ($PrP^{Sc}$). This $PrP^{Sc}$ is detergent insoluble and partially protease-resistant ($PrP^{res}$). The term $PrP^{res}$ is normally used to describe the protein detected after protease treatment, in techniques such as Western immunoblotting, and enzyme-linked immunosorbant assay using fresh/frozen tissue. Immunohistochemistry may performed with formalin-fixed tissues. Also, clinical signs of the BSE are one of the major diagnostic indicators. Recently, atypical forms (known as H- and L-type) of BSE have appeared in several European countries, Japan, Canada and the United States. An unusual case was also reported in a miniature zebu. The atypical BSE fall into two groups based on the relative molecular mass (Mm) of the unglycosylated $PrP^{res}$ band relative to that of classical BSE, one of the higher Mm (H-type) and the other lower (L-type). Both types have been detected worldwide as rare cases in older animals, at a low prevalence consistent with the possibility of sporadic forms of prion diseases in cattle. This raises the unwelcome possibility that vCJD could increase in the human population. Now, active surveillance program against BSE is going on in Korea. In regional veterinary service lab, ELISA is applied to screen the BSE in slaughter and confirmatory tests by Western immunoblotting and immunohistochemisty are carried out if there are positive or suspect in the screening test. Also, the ruminant feed ban is rigorously enforced. Removal of specified risk materials such as brain and spinal cord from cattle is mandatory process at slaughter to prevent the infected material from entering the human food chain.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.4
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• pp.509-518
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• 2002

The purpose of this study was to evaluate the bonding of compomer to deciduous dentin which is known to have been developed to improve the weak properties of glass ionomer cement and composite resin. 120 sound primary molars were used for the shear bond strength test and another 24 for the scanning electron microscopic evaluation. Each material was ailed into polyethylene mold attached to exposed dentinal surface($3{\times}4mm$ in diameter) of sample blocks. Shearbond strength was measured using Universal testing machine and data were analyzed statistically with Oneway-ANOVA and Scheffe test. Scanning electron microscopic observation was performed in order to evaluate the pattern of distribution and penetration of resin tags and hybrid layer. Compomer groups(II-V) showed significantly higher bond strength values than glass ionomer group(I)(p<.05). Etching-compomer groups(III, V) showed the significantly higher bond strength than non-etching compomer groups(II, IV)(p<.05), but slightly lower values than composite resin group(VI) with no statistically significant difference(p>.05). No significantly different bond strength was found between compomer groups of different bonding system(p>.05). Scanning electron micrographs showed more irregular distribution of short and thin resin tags in non-etching compomer groups(II, IV) whereas the more regular and intimate distribution of long and thick tags in etching compomer groups(III, V) and composite resin group(VI). The evaluation of hybrid layer also showed more regular formation of thicker layer in etching compomer groups(III, V). Based on the results of present study, the use of compomer as an esthetic restorative material for primary molars might be justified.

• Economic and Environmental Geology
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• v.41 no.6
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• pp.685-693
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• 2008

The rates of oxidative degradation of perchloroethene (PCE) and trichloroethene (TCE) using $KMnO_4$ solution were evaluated under the flow condition using a bench-scale transport experimental setup. Parameters which are considered to affect the reaction rates tested in this study were the contact time (or retention time), and the concentration of oxidizing agent. A glass column packed with coarse sand was used for simulating the aquifer condition. Contact time between reactants was controlled by changing the flow rate of the solution through the column. The inflow concentrations of PCE and TCE were controlled constant within the range of $0.11{\sim}0.21\;mM$ and $1.3{\sim}1.5\;mM$, respectively. And the contact time was $14{\sim}125$ min for PCE and $15{\sim}36$ min for TCE. The $KMnO_4$ concentration was controlled constant during experiment in the range of $0.6{\sim}2.5\;mM$. It was found that the reduction of PCE and TCE concentrations were inversely proportional to the contact time. The exact reaction order for the PCE and TCE degradation reaction could not be determined under the experimental condition used in this study. However, the estimated reaction rate constants assuming pseudo-1st order reaction agree with those reported based on batch studies. TCE degradation rate was proportional to $KMnO_4$ concentration. This was considered to be the result of using high inflow concentrations of reactant, which might be the case at the vicinity of the source zones in aquifer. The results of this study, performed using a dynamic flow system, are expected to provide useful information for designing and implementing a field scale oxidative removal process for PCE/TCE-contaminated sites.

• Journal of the Korea Organic Resources Recycling Association
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• v.25 no.2
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• pp.59-67
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• 2017

The aim of this experiment was to investigate the effect of air suction rate (SR) during the composting process of swine manure mixed with sawdust used as a bulking agent. In the 25 L composting reactors, the suction rate (SR) was at four different treatment levels (100%, 200%, 300%, 400%), and were fixed on the based on constant aeration rate into the composting mixtures. The temperature reached to thermophilic phase within 2 days and it was maintained up to the $5^{th}$ day of the composting process in all reactors and then gradually decreased to room temperature at the end of the composting process. The moisture content (MC, %) of the initial mixtures was 64.27%, and it was reduced to 38.4, 33.08, 14.59 and 11.93 in the different suction rate of 100%, 200%, 300%, 400%, respectively in the end process. During the composting, the level of pH was increased from 6.83 to 8.67 and it gradually decreased to 7.56 in 100% and 200%(SR). At the same time, the pH values were reduced only up to 8.19 at 300%, and 8.08 at 400%(SR), showing that suction strengths of 100% and 200% were the better option for composting than those of 300% and 400%. The total Kjeldahl nitrogen (TKN) of initial composts mixtures was 2.3% and were changed in 3.3, 3.1, 2.5, and 2.3% at the end of the composting period from the 100%-400% (SR) variations respectively. These results also indicated that 100% and 200% (SR) were more affected by the dry mass loss as $CO_2$ and water evaporation. The initial value of C/N ratio was 25.17 and were significantly reduced to 11.88, 11.97, 14.31, and 14.72 at the end of the experiment, respectively from the 100%-400% (SR) variations. These results suggest that the suction rate (SR) of 100% and 200% relative to constant air supply would be the optimal conditions to produce high-quality compost.