It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.
Objectives: Ethylene oxide (EtO) is classified as a human carcinogen, but EtO is still widely used to sterilize heat-sensitive materials in hospitals. Employees working around sterilizers are exposed to EtO after sterilization. The aim of the present study was to assess the exposure of EtO level, coupled with occupationally induced micronuclei from hospital workers. The influence of genetic polymorphisms of detoxifying genes (GSTT1 and GSTM1) and DNA repair genes (XRCC1 and XRCC3) on the frequencies of micronuclei in relation to exposure of EtO was also investigated. Methods: The study population was composed of 35 occupationally exposed workers to EtO, 18 student controls and 44 unexposed hospital controls in Korea. Exposure to EtO is measured by passive personal samplers. We analyzed the frequencies of micronuclei by performing cytokinesis-block micronucleus assay (CBMN assay) and GSTM1, GSTT1, XRCC1, and XRCC3 were also genotyped by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequencies of micronuclei in EtO exposure group, student controls and hospital controls were $18.00{\pm}7.73$, $10.47{\pm}7.96$ and $13.86{\pm}6.35$ respectively and their differences were statistically significant, but no significant differences according to the level of EtO were observed. There was a dose-response relationship between the frequencies of micronuclei and cumulative dose of EtO, but no significantly differences were observed. We also investigated the influence of genetic polymorphisms (GSTM1, GSTT1, XRCC1, and XRCC3) on the frequencies of micronuclei, but there were no differences in the frequencies of micronuclei by genetic polymorphisms. Conclusions: The frequencies of micronuclei in EtO exposure group was significantly higher than control groups. A dose-response relationship was found between the level of EtO exposure and the frequencies of micronuclei, but no statistically differences were observed. We also found that the frequencies of micronuclei were increased according to cumulative EtO level. There was no association of the genetic GSTM1, GSTT1, XRCC1, and XRCC3 state with the frequency of micronuclei induced by EtO exposure.
Roads are an indicator of anthropogenic activity causing ecosystem disturbances and often lead to habitat fragmentation, habitat loss, and habitat isolation. The Hallasan National Park(153.4$km^2$) on Jeju Island being distinguished for its unique geology, topography, and biota has also been designated as a core area of UNESCO Man and the Biosphere(MAB) Reserve. Although the high conservation value of this park has contributed to a rapid growth of tourists and road construction, landscape changes due to roads have not been examined yet. We used GIS systems to examine the fragmentation pattern caused by roads, in relation to its zonation, elevation, and vegetation. When a buffer was applied to roads(112m width for paved roads and 60m width for both legal and illegal trails), the park consisted of 100 fragments. The ten fragments generated after applying buffer to only paved roads and legal trails ranged from $0.002km^2$ to $38.2km^2$ with a mean of $14.2km^2$, and about 7% of both nature conservation zone and nature environment zone of the park were edge. Fragments in both east and west ends of the park and around the summit exhibited relatively high shape indices with means of 5.19(for 100 fragments) and 7.22(for 10 fragments). All five legal trails are connected to the pit crater of the mountain and vegetation changed from broadleaf forests and conifer forests to grasslands with elevation, consequently resulting in dramatic fragment size reduction in grasslands at high elevation, in particular above 1,400m, where endemic and alpine plants are abundant. These results show that in Hallasan National Park the risks of habitat deterioration and habitat loss due to fragmentation may be more severe in the nature conservation zone dominated by Baengnokdam than in the nature environment zone. Therefore, current road networks of the park appear to fall short of the goal of the national park for ecosystem conservation and protection. Considering that the entire Hallasan National Park also serves as a MAB core area, conservation efforts should focus, first of all, on park rezoning and road management to mitigate habitat fragmentation.
Background : It has been reported that nontuberculosis mycobacterium(NTM) isolates account for approximately 10% of patients with a positive Acid-Fast Bacilli(AFB) smear. Therefore, it is necessary to consider NTM pulmonary disease when such a positive test is encountered. The aim of this study was to evaluate the etiologies and clinical characteristics of patients with NTM pulmonary disease who had been treated at a national tuberculosis hospital. Methods : The NTM isolates were recovered from the sputum or bronchial washing specimens submitted to a clinical laboratory of National Masan TB Hospital from August 2002 to July 2003. All samples were identified using a polymerase chain reaction-restriction fragment length polymorphism analysis method, which amplifies the rpoB gene. The patients were diagnosed with NTM disease according to the American Thoracic Society diagnostic criteria. Results : One hundred NTM isolates were recovered from 57 patients. Of the 100 isolates, M. avium complex(MAC) was the most common species, which was found 55%(n=55) of patients, followed by M. abscessus(n=25), and M. fortuitum( n=9). 26(45.6%) patients had NTM disease. Twenty-six (45.6%) patients had NTM disease according to The American Thoracic Society classification. The main organisms involved in NTM disease were MAC(n=19, 73.1%) and M. abscessus(n=5, 19.2%). The pathogenic potential was 67.9% in M. intracellulare and 41.7% in M. abscessus. The predictive factors related to NTM disease were a positive sputum smear (OR 6.4, p=0.02) and the isolation of either MAC or M. abscessus(OR 6.9, p=0.007). Fifteen patients(57.7%) were cured. There were no significant factors associated with the treatment success. Conclusion : There was a relatively high proportion of NTM disease in NTM isolates and the common species were MAC and M. abscessus. The predictive factors for NTM disease were a positive sputum smear and the isolation of either MAC or M. abscessus.
Lee, Jung Yeon;Kim, Mi-Na;Chung, Hee-Jung;Jun, Kyung Ran;Choi, Hee Jin;Lee, Hyeyoung;Joung, Eun Young;Oh, Yeon-Mok;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Shim, Tae Sun
Tuberculosis and Respiratory Diseases
/
v.59
no.1
/
pp.39-46
/
2005
Background : Even though it has been suggested that low-colony, scotochromogen nontuberculous mycobacteria (NTM) are usually contaminants and not true pathogens, evidence for this hypothesis has not been provided. This study investigated the colony characteristics, organism identification, and clinical significance of low-colony scotochromogen. Methods : The laboratory cultured 6,898 respiratory clinical specimens for an examination of mycobacteria over a three-month period. A low-colony count was arbitrarily defined as ${\leq}20$ colonies. This study analyzed the recovery rate of the mycobacteria, the number of colonies and their gross characteristics, and their clinical significance. PCR-restriction fragment length polymorphism analysis was carried out to identify the NTM species. NTM pulmonary disease was defined according to the American Thoracic Society. Results : A total of 6,898 respiratory specimens for mycobacterium were cultured. Of these, 263 (3.8%) grew NTM, and 382 (5.5%) grew M. tuberculosis. Of the 263 cultured NTM specimens, 124 (47.1%) were scotochromogens. The smear-positive rate was significantly lower in these scotochromogens (4.8%) than in the non-scotochromogens (23.7%) (p<0.05). The most common isolates were M. gordonae (83/102, 81.4%) in the scotochromogens, and MAC (52/121, 43.0%) in the non-scotochromogens. Even though three out of 113 patients with a low-colony scotochromogen has been diagnosed with NTM pulmonary disease, the isolated scotochromogen was not considered to be the cause of the NTM disease but was just a contaminant. Conclusion : In this study, the most common isolate of a low-colony count scotochromogen was M. gordonae, which appeared to be contaminants and not true pathogens. Greater efforts in the quality control of a mycobacterium laboratory are needed in cases where there is a high recovery rate of low-colony count scotochromogen.
Journal of the Korea Academia-Industrial cooperation Society
/
v.18
no.12
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pp.575-580
/
2017
This study analyzed the effects of RGG box of hnRNP A1 on its subcellular localization and stabilization of hnRNP A1 over a three year period from October 2014. First, a 6R/K mutation in RGG box was generated, and pcDNA1-HA-hnRNP A1(6R/K) was constructed. The subcellular localization of hnRNP A1(6R/K) from the HeLa cells transfected with this plasmid DNA was analyzed by immunofluorescence microscopy. HA-hnRNP A1(6R/K) was found to exhibit nuclear and cytoplasmic fluorescence. The stability of hnRNP A1(6R/K) was checked by Western blot analysis using the expressed protein from the HeLa cells transfected with the pcDNA1-HA-hnRNP A1(6R/K). The results show that HA-hnRNP A1(6R/K) has a smaller size. These confirm that HA-hnRNP A1(6R/K) is localized both in the nuclear and cytoplasm, not because 6R/K mutation affects the nuclear localization of hnRNP A1, but because 6R/K mutation causes hnRNP A1(6R/K) to cleave at the mutation or near the mutation site. The cleaved protein fragment, which lacks the M9 domain (i.e. nuclear localization signal of hnRNP A1), did not exhibit nuclear fluorescence. This suggests that the arginines of RGG box in hnRNP A1 play an important role in stabilizing hnRNP A1. An analysis of the RNA-binding ability of hnRNP A1(6R/K) expressed and purified from bacteria will be a subsequent research project.
Lee, H.J.;Lee, S.H.;Cho, Y.M.;Yoon, H.B.;Jeon, B. K.;Oh, S.J.;Kwon, M.S.;Yoon, D.H.
Journal of Animal Science and Technology
/
v.46
no.6
/
pp.947-956
/
2004
The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of triglyceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cattle, we have directly sequenced the exon 5${\sim}$exon 6 region in the bovine LPL gene for discovery of single nucleotide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the polymorphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P<0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.
To evaluate the effects of chemical pretreatments of lignocellulosic biomass on enzymatic hydrolysis process, Populus euramericana was pretreated for 1 hr with 1% sulfuric acid ($H_2SO_4$) at $150^{\circ}C$ and 1% sodium hydroxide (NaOH) at $160^{\circ}C$, respectively. Before the enzymatic hydrolysis, each pretreated sample was subjected to drying process and thus finally divided into four subgroups; dried or non-dried acid pretreated samples and dried or non-dried alkali pretreated samples and chemical and physical properties of them were analyzed. Biomass degradation by acid pretreatment was determined to 6% higher compared to alkali pretreatment. By the action of acid ca. 24.5% of biomass was dissolved into solution, while alkali degraded ca. 18.6% of biomass. However, reverse results were observed in delignification rates, in which alkali pretreatment released 2% more lignin fragment from biomass to the solution than acid pretreatment. Unexpectedly, samples after both pretreatments were determined to somewhat higher crystallinity than untreated samples. This result may be explained by selective disrupture of amorphous region in cellulose during pretreatments, thus the cellulose crystallinity seems to be accumulated in the pretreated samples. SEM images revealed that pretreated samples showed relative rough and partly cracked surfaces due to the decomposition of components, but the image of acid pretreated samples which were dried was similar to that of the control. In pore size distribution, dried acid pretreated samples were similar to the control, while that in alkali pretreated samples was gradually increased as pore diameter increased. The pore volume which increased by acid pretreatment rapidly decreased by drying process. Alkali pretreatment was much more effective on enzymatic digestibility than acid pretreatment. The sample after alkali pretreatment was enzymatically hydrolyzed up to 45.8%, while only 26.9% of acid pretreated sample was digested at the same condition. The high digestibility of the sample was also influenced to the yields of monomeric sugars during enzymatic hydrolysis. In addition, drying process of pretreated samples affected detrimentally not only to digestibility but also to the yields of monomeric sugars.
Microcystin produced by cyanobacteria in surface waters, such as eutrophic lake and river, is a kind of serious environmental problems due to its toxicity to human and wild animals. Microcystin is synthesized nonribosomally by the large modular multi-functional enzyme complex known as microcystin synthetase encoded by the mcy gene cluster. Amplification of mcy genes by PCR from cultures and environmental samples is a simple and efficient method to detect the toxigenic Microcystis. In order to evaluate primers designed to detect toxic microcystin-producing strains, 17 cyanobacterial strains and 20 environmental samples were examined by PCR with 7 pairs of primers. Some microcystin-producing cyanobacteria were not detected with FAA-RAA, TOX4F-TOX4R and FP-RP primers. The fragment of unexpected size was amplified with NSZW2-NSZW1 primers in Microcystis strains isolated from the lakes in Korea. TOX1P-TOX1F primers failed in amplification of toxin-producing strains. Only MSF-MSR and TOX2P- TOX2F primers amplified the fragments of mcy genes from 11 strains of microcystin-producing Microcystis. The water samples taken from 20 lakes in Korea were analyzed by PCR using each of the primers. In all the water samples, cyanobacteria capable of producing microcystin were detected by the PCR with TOX2P-TOX2F primers. These results indicate that TOX2P-TOX2F primers are better than the other primers for detection of microcystin-producing Microcystis strains in Korea. The nucleotide sequences of mcy gene in Microcystis aeruginosa NIER10010 suggest genetic diversity of Korean isolates.
This paper investigates Buddhist sculptures of the Gulsan-sa School of Seon Buddhism, which was established by the priest Beomil(梵日) at Myeongju(溟州), present-day Gangneung(江陵) area, and prospered until the late Goryeo period. There are very few surviving Buddhist images at Gulsan2 temple-site showing the Buddhist art when Beomil or his disciples, Gaecheong(開淸) and Haengjeok(行寂), were active, except a set of huge stone pillars for temple-banners and a stone monk stupa. It is therefore necessary to focus on pieces of artworks in Gangneung and nearby areas to explore the characteristics of Buddhist art of Gulsan-sa from the late Silla to early Goryeo period. For example, the stone relief seated Buddha image, one of Four Buddhas of Four Directions, and a stone relief of standing Guardian King both from the stone pagoda at Mujin temple-site(無盡寺址) can be compared with stupa reliefs at the capital Gyeongju area in their high quality of carving. The stone octagonal base showing seven lions on each of seven sides at Boheon-sa(普賢寺) demonstrates that it was made as a part of an octagonal lotus pedestal for a Vairocana Buddha now lost. Since Boheon-sa was erected by Gaechung, a disciple of Beomil as a branch of Gulsan-sa, it is fair to assume that the Vairocana image at Boheon-sa might have been closely related to the Buddhist images at Gulsan-sa in its style and iconography. The stone seated Bodhisattva from Hansong temple-site(寒松寺址) displays a benign face, exquisite necklace, and exotic iconography in its hand gesture and high cylindrical crown. The stone seated Buddha at Cheonghak-sa(靑鶴寺), brought from a temple-site where fragments of roof-tiles with the inscription of Heukam-sa(黑岩寺) were discovered, displays the late Silla and early Goryeo period. Heukam-sa seems to have been related to Gulsan-sa or have been one of the branches of Gulsan-sa. Extant fragments of artworks at Myeongju implicate the high quality of Buddhist art of Gulsan-sa in its iconography and style as well as unique features of Gulsan-sa Seon School.
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