• Title/Summary/Keyword: Colony development

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Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis (면역체 분석을 위한 탄저균 유전자 발현 라이브러리의 구축)

  • Park, Moon-Kyoo;Jung, Kyoung-Hwa;Kim, Yeon-Hee;Rhie, Gi-Eun;Chai, Young-Gyu;Yoon, Jang-W.
    • 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.

INHIBITORY EFFECT OF Er:YAG LASER ON THE GROWTH OF STREPTOCOCCUS MUTANS (Er:YAG 레이저 조사가 Streptococcus mutans의 증식억제에 미치는 효과)

  • Song, Gwang-Chul;Lee, Chang-Seop;Lee, Sang-Ho;Lee, Nan-Young
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.15-24
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    • 2003
  • The purpose of this study is to investigate the sterilization effect of Er:YAG laser against the intraoral acid producing bacterium, S. mutans, by irradiating the culture solution containing S. mutans KCTC 3065 with Er:YAG laser having a $650{\mu}m$ diameter beam through the non-contact method. We obtained the following results after examining the temperature changes of the culture solution, numbers of bacterial colonies, and acid-producing ability and attaching ability on teeth by measuring the amount of extracellular polysaccharide produced by S. mutans. The number of bacterial colony was decreased in $10{\mu}l$ culture solution irradiated with laser in overall compared to the control solution. The number decreased as the irradiation intensity and pulse repetition rate were larger and as the exposure time was increased. However, it did not change significantly in $100{\mu}l$ culture solution compared to the control solution. Although the acid-producing ability of S. mutans was inhibited for a certain duration after laser irradiation in 10r1 bacterial culture solution, it did not change in $100{\mu}m$ solution compared with the control solution. The amount of extracellular polysaccharide synthesized by S. mutans was partially decreased through laser irradiation in $10{\mu}m$ culture solution but did not change in $100{\mu}m$ culture solution. Based on these findings, we concluded that Er:YAG laser has an sterilization effect on S. mutans in which we presume that the mechanism is through the heat effect rather than the mechanical effect from development of ultrasound.

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Changes of Street Patterns in Central Part of Taegu City (大邱市 都心部의 街路網 變化)

  • Choi, Seok-Joo
    • Journal of the Korean Geographical Society
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    • v.31 no.3
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    • pp.593-612
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    • 1996
  • This study concern with the changes of street from the Choseon Dynasty to present days around Old-Boundary in Taegu, analyzing the backgrounds of change factors and development trends of the Taegu City. The basis element of a city structure is the street. Therefore, in this study, a chage of street space of a city was investigted. Historically, Taegu was a walled city and had a Mono-nucleus which was restricted by the castle, and served as a starting point of formation of spatial structuure. The form of the artery street took a "T" pattern, othe streets were formed in irregular shapes. As the city grew gradually, the castle was removed on account of diversification in traffic network, change of socio-economic organization in traffic network, change of socio-economic organization, formation of industrial bases and functional distribution. CBD of this city has been located within the area surrounded by these streets. This is a kind of general pattern of traditional walled cities through the world in both Western and Oriental societies. A s the begining of this centry, a 'Dark Ages' descended upon Korea because the country was under the Japanese-Korean Annexation, and, throughout this period, the urban planning was planned exclusively for Japanese. The street pattern within residential areas of Korea took the maze type, in contrast with Japanese residential areas which showed grid pattern of streets. This is another general pattern of almost of all colonial cities especially in Asia. High class residential areas were planned and built by Japanese, and they were located within 5-10 minutes' on-foot distance from the CBD hard core. This high prestige has continued until the 1980s when it occurred land use succession which commerical functions invaded into residential areas. Back in the colonial period, there was a between two hetrogeneous groups due to the fact that the Japanese lived mainly oriented the new railway system but that Koreans still lived along the old highway system which ran through the Korea Peninsula. Street netwook formed in the above process has maintained its shape without great changes after the liberation form the Japanese Colony. Taegu has, accordingly, developed ring-radial network system which has been a combination of radial and ring facilities. The present conditions of street patterns in Taegy mainly depend on 4 rings and 8 radius, with grid pattern street able to be found in Old Boundary.

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The Changes of Intestinal Normal Flora in Neonates for Seven Days Postnatally (정상 신생아의 대변에서 생후 1주일 동안 장내세균총의 변화)

  • Sung, Nam-ju;Lee, Seung Gue;Kim, Me Jin;Kim, Young Ho;Yang, Seung;Hwang, Il Tae;Jung, Ji A;Lee, Hae Ran;Kim, Jae-Seok
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.2
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    • pp.162-168
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    • 2006
  • Purpose: Microbial colonization of the intestine begins just after birth and development of the normal flora is a gradual process. The first bacteria colonizing the intestine in newborns are Staphylococcus, Enterobacteriaceae and Streptococcus. For several days after birth, the number of Bifidobacterium spp. increase. The aim of this study was to investigate the changes of microflora for seven days postnatally in neonatal stool. Methods: Fifteen neonates (breast : formula : mixed feeding 1 : 8 : 6, vaginal delivery : cesarean section 3 : 12) who were born at the Kangdong Sacred Heart Hospital, Hallym University were enrolled. First meconium and stools of postnatal 1-, 3-, and 7-day were innoculated. Blood agar plates for total aerobes, trypton bile X-glucuronide agar for E. coli, phenylethyl alcohol agar for gram positive anaerobes, MRS agar for Lactobacillus spp., bifidobacterium selective agar for Bifidobacterium spp. and cefoxitin-cycloserine-fructose agar for Clostridium difficile were used in the general incubator ($CO_2$ free incubator), $CO_2$ incubator or the anaerobic chamber for 48 or 72 hours at $37^{\circ}C$ and then colony forming units were counted. Results: No microflora was identified in the first meconium. Total aerobes, E. coli, and gram positive anaerobes were significantly increased with advancing postnatal days. In only one baby, Lactobacillus acidophilus was detected $2{\times}10^5CFU/g$ in the seven-day stool. Bifidobacterium spp. was detected in two babies. Clostridium difficile was not detected during the seven days. There were no significant differences in the bowel flora depending on the delivery pattern and feeding method. Conclusion: This study shows many changes in the intestinal normal flora in neonatal stool during seven days postnatally. If these findings are confirmed with larger studies, the data may be preliminary findings to support use of probiotics in neonates.

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Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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The Anti-Sticking Effect of Mixture of Trisodium Phosphate and Citric Acid on Oral Streptococcus species (구강 내 사슬알균 종들에 대한 제3인산나트륨과 구연산의 탈부착 효과)

  • Jung, Choong-Hyun;Cho, Hyung-Hun;Choi, Gwang-Ju;Kang, Seung-Yong;Yang, Nam-Woong
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.289-292
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    • 2008
  • Trisodium phosphate 12 hydrate and citric acid monohydrate mixture showed the strong anti-sticking effect on Streptococcus mutans, Streptococcus mitis, and Streptococcus salivarius, which are adhered to glass beads. Each Streptococcus species was shaking-cultured in brain heart infusion broth containing three glass beads. After 18 hr, glass beads were slightly washed into normal saline by three-pin-pointed pincette. Each three glass-beads set was put into reagent -containing tubes, which have 40 mg of bits of weighing paper for gaining brushing effect as similar as brushing one's teeth. The tubes were shaken by vortex mixer for 10 min except non-oral microbe, Streptococcus agalactiae (5 min). The samples were colony-counted by serial agar dilution method. Experiment was repeated three times for each Streptococcus species. The relative ratios of bacterial de-adherence by reagents were calculated in comparison with normal saline control. The de-adherence degree of citric acid-trisodium phosphate-saline mixture (CTS, pH 6.0) against Streptococcus mutans came to an average of 12.5 times compared with normal saline control. Trisodium-saline (TS, pH 8.4) showed the average of 7.5 times, and citric acid-saline (CS, pH 4.6) showed 6.0 times compared to the control group. The bacterial de-adherence degree against Streptococcus salivarius was each 7.2,2.6 and 2.8 times in above reagent sequence in comparison with saline control. CTS and TS showed 2.4 and 3.4 times of anti-sticking effect on Streptococcus mitis respectively, but CS had no anti-sticking effect on this bacterium. CTS, TS and CS showed 0.7, 0.6, and 0.6 times on non-oral microbe, Streptococcus agalactiae, separately compared with saline control. These results show that oral Streptococcus mutans, Streptococcus salivarius, and Streptococcus mitis, which are causative of dental caries or subacute endocarditis, may be easily removed from oral cavity by CTS mixture. It is conceivable that our experimental results will enable the development of a new conceptive toothpaste to prevent dental caries or subacute endocarditis after drawing teeth.

A History of Termite Control and Improvements to Prevent Termites in Wooden Architectural Heritage (국내외 흰개미 방제 기술의 발달 과정과 목조건축문화재의 흰개미 피해 저감을 위한 방안)

  • LEE, Sangbin;IM, Ikgyun;KIM, Sihyun
    • Korean Journal of Heritage: History & Science
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    • v.54 no.2
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    • pp.194-215
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    • 2021
  • Termites are important decomposers in the ecosystem. They are also economically significant structural pests. In this study, we reviewed the developments of termite control and recent research on termite management to provide information on the prevention and control of termites. In Korea, most of the damage to wooden historical buildings is caused by subterranean termites. Reticulitermes speratus kyushuensis is the main species, which is widely found throughout the country. In the early 1900s, inorganic insecticides, such as arsenic dust, were used for termite control. After the synthesis of chlorinated hydrocarbon pesticide in the 1940s, it was widely utilized and demonstrated high termite control efficacy. However, chlorinated hydrocarbon insecticides were later banned, disappearing from markets after reports emerged concerning environmental contamination and toxicity to humans. Therefore, the termite control industry sought a new termiticide; hence many pesticides were utilized for termite control. Organophosphate (1960s), carbamate (1970s), pyrethroid, and insect growth inhibitor (1980s) were newly synthesized and adopted. In the 1990s, the first commercial baits using chitin synthesis inhibitors (CSI) were developed, providing a means to eliminate an entire colony of subterranean termites around a structure. Many studies have been carried out on soil termiticides (liquid termiticides) and CSI baits to increase their efficacy, and different baits such as aboveground bait stations, fluid bait, and high-durability bait were also developed in the 2000s. In addition, the paradigm of termite control has shifted from localized treatments using soil termiticides to area-wide pest management using CSI baits to create termite-free zones and protect buildings over time. Termite infestations in wooden historical buildings in Korea have been reported since 1980, and considerable attention was drawn in the 1990s when several UNESCO world heritages such as the Jongmyo Shrine and the Janggyeong Panjeon Depositories of Haeinsa Temple were infested by subterranean termites. Since then, a survey of termite infestation in wooden architectural heritage has been conducted, and the National Research Institute of Cultural Heritage and Heritage Care Program regularly monitors those properties. Finally, we suggest termite management using primarily CSI baits, selective application of various soil treatments applied to the object, foundation soil treatment, research and development of durable termite baits, application of area-wide programs for wooden-building complexes, application of integrated termite management (ITM), and regular education for owners and managers to prevent and reduce termite damage.

Pro-inflammatory Cytokine Expression Through NF-${\kappa}B/I{\kappa}B$ Pathway in Lung Epithelial Cells (폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로에 의한 염증매개 사이토카인의 발현)

  • Park, Gye-Young;Lee, Seung-Hee;HwangBo, Bin;Yim, Jae-Joon;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.332-342
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    • 2000
  • Background : The importance of pro-inflammatory cytokines, especially tumor necrosis factor $\alpha$ (INF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$), have been extensively documented in the generation of inflammatory lung disease. Lung epithelial cells are also actively involved in initiating and maintaining inflammation by producing pro-inflammatory mediators. Understanding the mechanism of pro-inflammatory cytokine expression in lung epithelial cells is crucial to the development of new therapeutic modalities for inflammatory lung disease. Transcription of most pro-inflammatory cytokines is dependent on the activation of NF-${\kappa}B$. However, the relationship between pro-inflammatory cytokine expression and NF-${\kappa}B/I{\kappa}B$ pathway in lung epithelial cells is not clear. Methods : BEAS-2B, A549, Na-H157, NCI-H719 cells were stimulated with IL-$1{\beta}$ or TNF-$\alpha$ at various times, and then IL-8 and TNF-$\alpha$mRNA expressions were assayed by Northern blot analysis. IL-$1{\beta}$ or TNF-$\alpha$-induced NF-${\kappa}B$ activation was assessed by the nuclear translocation of p65 NF-${\kappa}B$ subunit. The degradation of $I{\kappa}B{\alpha}$ and $I{\kappa}B{\beta}$ by IL-$1{\beta}$ or TNF-$\alpha$stimulation was assayed by Western blot analysis. The phosphorylation of $I{\kappa}B{\alpha}$ was evaluated by Western blot analysis after pre-treating cells with proteasome inhibitor followed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation. The basal level of IKK $\alpha$ expression was evaluated by Western blot analysis. Results: $I{\kappa}B{\alpha}$ and $I{\kappa}B{\alpha}$ was rapidly degraded after 5 minutes of incubation with IL-$1{\beta}$ or TNF-$\alpha$ in BEAS-2B, A549, and NCI-H157 cells. The activation of NF-${\kappa}B{\alpha}$ and the induction of IL-8 and TNF-$\alpha$ mRNA expression were observed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in these cells. In contrast, neither the changes in NF-${\kappa}B/I{\kappa}B$ pathway nor IL-8 and TNF-$\alpha$mRNA expression was induced by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in NCI-H719 cells. IL-$1{\beta}$ and TNF-$\alpha$-induced $I{\kappa}B$ phosphorylation was observed in BEAS-2B, A549, and NCI-H157 cells, but not in NCI-H719 cells. The basal level of IKK$\alpha$ expression was not different between cell. Conclusion : NF-${\kappa}B/I{\kappa}B$ pathway plays an important role in the expression of pro-inflammatory cytokine in most lung epithelial cells. The absence of the effect on NF-${\kappa}B/I{\kappa}B$ pathway in NCI-H719 cells sæms to be due to the defect in the intracellular signal transduction pathway upstream to IKK.

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