• 대한치과보철학회지
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• 제41권2호
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• pp.207-222
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• 2003

As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex ; tissue stimulation, skin allergy, hypersensitivity cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Niresistance in oral microorganisms. The present study was undertaken to check whether use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the patients wearing Ni-Cr prosthesis. The isolated bacteria were tested fir their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several biochemical, molecular-biological tests. Performed tests were : measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy prosthesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergoviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin, However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggests that there is no homology between the previously known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.

• 대한치과보철학회지
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• 제37권6호
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• pp.741-755
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• 1999

As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex : tissue stimulation, skin allergy, hypersensitivity, cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Ni-resistance in oral microorganisms. The present study was undertaken to check wheather use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the pateints wearing Ni-Cr prosthesis. The isolated bacteria were tested for their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several bio-chemical, molecular-biological tests. Performed tests were ; measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows: 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy pros-thesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergeviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin. However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggest that there is no homology between the previousely known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.

• 한국환경과학회지
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• 제6권3호
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• pp.249-258
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• 1997

Mercury- and cadmium-resistant bacteria were Isolated from an Industrial complex wastewater of Taejon area. All of them were motile, gram negative rods. In the results of physicochemical test and VITEK card test. HM1 was Identified with Achetobacter cd- coaceucus, CM3 was Identified 65 Commonas acidovorns, HM2, HM3, CMI , and CM4 were Pseudomonas sp., but HM4 and CM2 were unidenteed. They were tested for subceptlbility to 14 heavy metals. Mercury-resistant bacteria(HM1, HM2, HM3, and HM4) were sensitive to low concentration(100~400ppm) of $Cd^{2+}$, $Co^{2+}$. $Zn^{2+}$, and Ni2+, while cadmium-resistant bacteria(CM1, CM2, CM3, and CM4) showed resistance up to the high concentration(600~ 1, 200ppm) of these metal loons. As a result of resistance spectrum test of mercury-resistant bacteria, HM1 was broad-spectrum strain, HM2. HM3, and HM4 were narrow-spectrum stratas. Transmission electron microscopic examination of cell wall of HM1 culture grown with and without 100ppm of $HgCl_2$ showed remarkably merphological abnormalities. In the result of atomic absorption spectrometric analysis of cadmium-resistant bacteria grown at 200ppm of $CdCl_2$ for 6h, all of them accumulated cadrnium(14ppm~57ppm) In cell. In cadinium-resistant bacteria, CM1, CM2, and CM4 were spared from the Inhibitory effect of $Cd^{2+}$ by the addition of $Mn^{2+}$, CM4 were also spared from the Inhibitory effect of $Cd^{2+}$ by the addition of $Mn^{2+}$ as well as $Zn^{2+}$.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.274-282
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• 2019

Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

• 대한소아치과학회지
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• 제27권2호
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• pp.274-282
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• 2000

소아의 유구치 수복에 흔히 사용하는 stainless steel crown은 주성분으로 $70\sim90%$의 니켈과 $5\sim15%$의 크롬을 함유하고 있다. 이 중 니켈 자체가 일으킬 수 있는 allergy반응이나 발암성등의 부작용이 성인에서는 활발하게 연구되고 있으며, 특히 니켈이 포함된 합금을 이용한 보철물 주변에 병원성이 있는 니켈내성 Enterococci가 존재한다는 보고가 있어 성인과는 구강환경이 다른 소아에서도 stainless steel crown 주변에 니켈내성균주가 존재하는지를 확인하고자 이 실험을 시행하였다. 유구치에 stainless steel crown(Anatom primary crown, Sankin)을 6개월 이상 장착한 환자 15명과 유구치 협면에 치아우식증이 없는 환자 15명의 치은열구액에서 채취한 시료를 BHI 한천배지, 니켈배지 및 Bile-esculin azide(BEA) 한천 배지에 도말하였다. BEA한천배지에서 자란 enterococci를 니켈내성정도를 확인하기위해 3, 5, 10, 30, 50mM 니켈배지에 접종한후 성장상태를 관찰하였다. 실험군에서는 BHI 한천배지에서 507,350개의 균주가 분리되었으며, 이 중 니켈내성세균(3mM)은 53,864 균주였다. 대조군에서는 BHI 한천배지에서 414,590 균주가, 니켈내성세균(3mM)은 37,523 균주가 분리되었다. BEA한천배지에서는 실험군의 경우 95개가 분리되었으며, 대조군에서는 20개의 균주만이 분리되었다. BHI 한천배지에서 분리된 총 균주 수를 100으로 놓았을 때 3mM 니켈배지에서 분리된 균주는 실험군에서는 10.62%, 대조군에서는 9.05%를 차지하여 두 군간에 뚜렷한 차이는 없었으며, 3mM 니켈배지에서 자란 총 균주 수와 BEA 한천배지에서 배양된 enterococci를 3mM 니켈배지로 옮겨 배양했을 때와 비교하면, 실험군에서 0.13%, 대조군에서 0.03%로 모두 1%이하의 분포를 차지하였다.

• Ocean and Polar Research
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• 제25권1호
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• pp.21-30
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• 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.

• 미생물학회지
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• 제30권3호
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• pp.199-206
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• 1992

Six isolated strains degrading aniline were selected, identified and designated as pseudomonas putida K6, Pseudomonas acidovorans K82, Achromobacter gr. D. V. K24, Achromobacter xylosocidans K4, Moraxella sp. K21 and Moraxella sp. K22. All of them degraded 1000 ppm aniline completely within 30 to 36 hours. Most of these strains are resistant to antibiotics more than one, but Moraxella sp. has not any antibiotic marker tested. Most strains except for P. acidovorans K82 were shown to have resistance to the heavy metal ions such as Ni, Cu, Li, Ba, Co, etc. but not to Hg to which only P. putida K6 was resistant. M. sp. K21 was capable of degrading aniline to a maximum concentration of 2500 ppm without any repression. The incubation of the cell in limited pH ranges (4-8) had no great effect on aniline degradation. The addition of bactopeptone to the minimal media promoted the speed of aniline degradation, but the addition of glucose rather repressed the rate of aniline degradation. Through enzyme assay, A. gr. D. V. K 24 was shown to degrade aniline through artho-pathway and formed .betha.-ketoadipate as intermediate metabolite.

• 미생물학회지
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• 제30권2호
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• pp.88-95
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• 1992

유전공학 기법으로 번형시킨 Km' plasmid 들의 전이를 30.deg.C 의 Luris-Bertani broth 에서 실시하였으며, 그 전이 빈도에 대한 donor 와 recipient 의 균주와 함께 pH 의 영향을 연구하였다. GEM 균주들과 NI 균주의 Km'plasmid 의 전이빈도는 recipient 가 MTI 일 경우 pH 7 에서 약 $10^{-5}$ 로 비슷하였으나 CS 균주인 DKC601 에서는 Km'plasmid 의 전이빈도가 $2.2 * 10^{-7}$ 로 훨씬 떨어졌다. 그리고 recipient 가 lab. strain (E. coli HB101) 인 경우에도 Km'plasmid 의 전이경향은 GEM 균주들과 NI 균주사이에 큰차이가 없었다. 또 어느 균주의 경우에나 pH 7 일 때의 전이 빈도가 $10^{-5}$ 정도로 가장 높게 나타났으며, pH 5 와 pH 9 에서는 그보다 조금 떨어졌다. 그러나 E. coli C600을 host 로 하여 제조한 CS 균주(DKC601)에서는 Km'plasmid 의 전이 빈도가 다른 GEM 균주들에 비하여 $10^{2}$ $-10^{3}$ 배 낮았으며 pH 4와 9 에서 6 시간의 conjugation 후에도 전이가 전혀 일어나지 않았다. Conjugant 에서의 plasmid의 재배열은 lab, strain 을 recipient 로 했을 경우에는 별로 없었지만, NI 균주를 recipient 로 했을 경우에 많이 나타났으며, 특히 donor 가 NI 균주보다 GEM 균주인 경우에 더욱 다양하였다. 그러나 pH 에 따른 타이는 그렇게 크지 않았다.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1119-1128
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• 2015

1-Aminocyclopropane-1-carboxylate (ACC) deaminase, which is encoded by some bacteria, can reduce the amount of ethylene, a root elongation inhibitor, and stimulate the growth of plants under various environmental stresses. The presence of ACC deaminase activity and the regulation of ACC in several rhizospheric bacteria have been reported. The nitrogen-fixing Pseudomonas stutzeri A1501 is capable of endophytic association with rice plants and promotes the growth of rice. However, the functional identification of ACC deaminase has not been performed. In this study, the proposed effect of ACC deaminase in P. stutzeri A1501 was investigated. Genome mining showed that P. stutzeri A1501 carries a single gene encoding ACC deaminase, designated acdS. The acdS mutant was devoid of ACC deaminase activity and was less resistant to NaCl and NiCl₂ compared with the wild-type. Furthermore, inactivation of acdS greatly impaired its nitrogenase activity under salt stress conditions. It was also observed that mutation of the acdS gene led to loss of the ability to promote the growth of rice under salt or heavy metal stress. Taken together, this study illustrates the essential role of ACC deaminase, not only in enhancing the salt or heavy metal tolerance of bacteria but also in improving the growth of plants, and provides a theoretical basis for studying the interaction between plant growth-promoting rhizobacteria and plants.

• 한국환경보건학회지
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• 제37권4호
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• pp.279-288
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• 2011

Objectives: This study investigated the levels and components of floor-settled dust in two elementary schools located at different sites (one near the Shihwa industrial complex and the other in a rural area) in order to evaluate the amounts of trace metal elements (As, Cd, Co, Cr, Cu, Ni, Pb and Zn) and microorganisms. Methods: Over twenty settled-dust samples were collected from the two elementary schools. Trace metal elements were extracted from the dust using hydrochloric acid and nitric acid, and the amounts were measured by ICP-OES. Microbiological analysis was performed by bacterial culturing using R2A medium and denaturing gradient gel electrophoresis (DGGE). Results: The results showed that the amounts of three metal elements (Cr, Pb, and Zn) were significantly different between the schools (${\alpha}$=0.05, p<0.04). In addition, microbial communities in each school were highly correlated with one another. Among the identified microorganisms, a number of potentially opportunistic microorganisms, including antibiotic-resistant bacteria such as Acinetobacter baumannii, were found. Conclusions: This study will provide preliminary data for assessing levels and types of chemical and microbiological agents in elementary schools and for further evaluating human health risks associated with the agents.