• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.27-28
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• 2014

Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

• Journal of Yeungnam Medical Science
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• v.14 no.2
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• pp.359-369
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• 1997

We studied the expression of the cell surface antigen associated with myeloid and lymphoid leukemias on bone marrow or peripheral blood blast cells from 153 leukemic patients including 61 cases of acute myelogenous leukemias(AML), 46 of acute lymphocytic leukemias(ALL) and 12 of acute leukemias. They were analyzed by direct or indirect immunofluorescence method for reactivity with the monoclonal antibodies to B cells(CD10, CD19, SmIg), T cells(CD2, CD5, CD7, CD3, CD4, CD8), myeloid antigen(CD13, CD14, CD33, CD61) and a nonspecific antigen, HLA-DR. Lymphoid associated markers detected on AML is CD7 32.8%, CD10 14.8%, CD5 13.1%, CD2 6.6% and CD19 1.6%. TdT was positive in 4.9% of AMLs. Hybrid leukemias were 8 cases out 61 AML cases and were mainly composed of monocytic lineage, M4 and M5a. Myeloid markers detected in ALL were CD13 2.2% and CD33 2.2%. In this study, immunologically classified ALLs were composed of 65.2% of CALLA (+) B precursor type, 10.9% of CALLA (-) B precursor pattern, 8.7% of T cell type, 2.2% of B cell type, 4.5% of mixed lymphoid lineage(B&T), 2.2% of undifferentiated leukemia, and 6.5% of hybrid leukemia. Twelve cases of acute leukemias ware finally diagnosed to be 5 cases of hybrid leukemia, 3 cases of B lineage, 3 case of T lineage and 1 case of mixed lymphoid(B&T) leukemia. In summary, we think the best method for typing acute leukemias is by using a combination of FAB classification and immunophenotying.

• Journal of Dental Rehabilitation and Applied Science
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• v.33 no.3
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• pp.178-188
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• 2017

Purpose: We compared the effects of newly developed diode laser (Bison 808 nm Diode laser) on the treatment of peri-implantitis with conventional products (Picasso 810 nm Diode laser) by comparing the surface temperature of titanium disc and bacterial sterilization according to laser power. Materials and Methods: The titanium disc was irradiated for 60 seconds and 1 - 2.5 W using diode laser 808 nm and 810 nm. The surface temperature of the titanium disc was measured using a temperature measurement module and a temperature measurement program. In addition, in order to investigate the sterilizing effect according to the laser power, 808 nm laser was irradiated after application of bacteria to sandblasted large-grit acid-etched (SLA) and resorbable blast media (RBM) coated titanium discs. The irradiated disks were examined with scanning electron microscopy. Results: Both 808 nm and 810 nm lasers increased disk surface temperature as the power increased. When the 810 nm was irradiated under all conditions, the initial temperature rise rate, the descending rate, and the temperature change before and after was higher than that of 808 nm. Disk surface changes were not observed on both lasers at all conditions. Bacteria were irradiated with 808 nm, and the bactericidal effect was increased as the power increased. Conclusion: When applying these diode lasers to the treatment of peri-implantitis, 808 nm which have a bactericidal effect with less temperature fluctuation in the same power conditions would be considered safer. However, in order to apply a laser treatment in the dental clinical field, various safety and reliability should be secured.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.15-21
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• 2011

We developed 14 transgenic lines of Chinese cabbage (Brassica rapa) harboring the T-DNA border sequences and CryIAc1 transgene of the binary vector 416 using Agrobacterium tumefaciens-mediated DNA transfer. Six lines had single copy cryIAc1 gene and four of them contained no vector backbone DNA. Of the left border (LB) flanking sequences six nucleotides were deleted in transgenic lines 416-2 and 416-3, eleven nucleotides in line 416-9, and 65 nucleotides including the whole LB sequences in line 416-17, respectively. And we defined 499 bp of genomic DNA (gDNA) of transformed Chinese cabbage, and blast results showed 96% homology with Brassica oleracea sequences. PCR with specific primer for the right border (RB) franking sequence revealed 834 bp of PCR product sequence, and it was consisted of 3' end of cryIAc1, nosterminal region and 52 bp of Chinese cabbage genomic DNA near RB. RB sequences were not found and the 58 nucleotides including 21 bp of nos-terminator 3' end were deleted. Also, there were deletion of 10 bp of the known genomic sequences and insertion of 65 bp undefined genomic sequences of Chinese cabbage in the integration site. These results demonstrate that the integration of T-DNA can be accompanied by unusual deletions and insertions both in transgenic and genomic sequences.

• Journal of Life Science
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• v.14 no.4
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• pp.650-656
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• 2004

Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

• Applied Chemistry for Engineering
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• v.28 no.1
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• pp.101-111
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• 2017

Mineral carbonation is a technology in which carbonates are synthesized from minerals including serpentine and olivine, and industrial wastes such as slag and cement, of which all contain calcium or magnesium when reacted with carbon dioxide. This study aims to develop the mineral carbonation technology for commercialization, which can reduce environmental burden and process cost through the reduction of carbon dioxide using steel slag and the slag reuse after calcium extraction. Calcium extraction was conducted using NH4Cl solution for air-cooled slag and convert slag, and ${\geq}98%$ purity calcium carbonate was synthesized by reaction with calcium-extracted solution and carbon dioxide. And we conducted experimentally to minimize the quantity of by-product, the slag residue after calcium extraction, which has occupied large amount of weight ratio (about 80-90%) at the point of mineral carbonation process using slag. The slag residue was used to replace silica sand in the manufacture of cement panel, and physical properties including compressive strength and flexible strength of panel using the slag residue and normal cement panel, respectively, were analyzed. The calcium concentration in extraction solution was analyzed by inductively coupled plasma optical emission spectrometer (ICP-OES). Field-emission scanning electron microscope (FE-SEM) was also used to identify the surface morphology of calcium carbonate, and XRD was used to analyze the crystallinity and the quantitative analysis of calcium carbonate. In addition, the cement panel evaluation was carried out according to KS L ISO 679, and the compressive strength and flexural strength of the panels were measured.

• Journal of the Korea institute for structural maintenance and inspection
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• v.19 no.5
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• pp.104-111
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• 2015

This study shows the mechanical properties of alkali-activated slag cement (AASC) synthesized using sulfate with NaOH solution. The used sulfates were calcium sulfate ($CaSO_4$, denoted CS) and sodium sulfate ($Na_2SO_4$, denoted SS). The replacement ratio of sulfates was 2.5, 5.0, 7.5 and 10.0% by weight of slag. NaOH solution of 2M and 4M concentration was used. A sample was activated with sulfate and activated with blended activator (blending NaOH solution with sulfate) respectively. 24 mix ratios were used and the water-binder weight ratio for the test was set 0.5. This research carried out the compressive strength, flexural strength, ultrasonic pulse velocity (UPV), absorption and X-ray diffraction (XRD). In the case of samples with CS, sample with 7.5% CS, sample with 2M NaOH+5.0% CS and sample with 4M NaOH+5.0% CS showed the good performance in the strength development. In the case of samples with SS, sample with 10.0% SS, sample with 2M NaOH+7.5% SS and sample with 4M NaOH+2.5% SS obtained good performance in strength. The results of UPV and water absorption showed a similar tendency to the strength properties. The XRD analysis of samples indicated that the hydration products formed in samples were ettringite, CSH and silicate phases. In this study, it is indicated that when compared to the use of sulfate only, the use of both sulfate and NaOH solution makes mechanical properties of AASC better.

• Journal of the Korean Orthopaedic Association
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• v.56 no.3
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• pp.234-244
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• 2021

Purpose: To date, studies of firearm and explosive injuries in the Korean military have been limited compared to its importance. To overcome this, this study examined the characteristics of musculoskeletal damages in soldiers who have suffered firearm and explosive injuries over the past four years. Materials and Methods: From January 2015 to July 2019, military forces who had suffered musculoskeletal injuries from firearms or explosive substances were included. The medical records and radiographs were reviewed retrospectively, and telephone surveys about Short Musculoskeletal Functional Assessment (SMFA) for this group were conducted. To compare the functional outcomes, statistical analysis was performed using a t-test for the types of weapons, and ANOVA for others. Results: Of the 61 patients treated for firearms and explosives injuries, 30 patients (49.2%) were included after undergoing orthopedic treatment due to musculoskeletal injury. The average age at injury was 26.4 years old (21-52 years old). The number of officers and soldiers was similar. Eleven were injured by gunshot and 19 by an explosive device. Sixteen were treated in the Armed Forces Capital Hospital and 10 at private hospitals. More than half of the 16 patients (53.3%) with a fracture had multiple fractures. The most common injury site was the hand (33.3%), followed by the lower leg (30.0%). There were 14 patients (46.7%) with Gustilo-Anderson classification 3B or higher who required a soft tissue reconstruction. Fifteen patients agreed to join the SMFA survey for the functional outcomes. Between officers and soldiers, officers had better scores in the Bother Index compared to soldiers (p=0.0045). Patients treated in the Armed Forces Capital Hospital had better scores in both the Dysfunction and Bother Index compared to private hospitals (p=0.0008, p=0.0149). Conclusion: This is the first study to analyze of weapons injuries in the Korean military. As a result of the study, the orthopedic burden was high in the treating patients with military weapon injuries. In addition, it is necessary to build a military trauma registry, including firearm and explosive injuries, for trauma treatment evaluation and development of military trauma system.