• Korean Journal of Soil Science and Fertilizer
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• v.39 no.6
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• pp.409-414
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• 2006

Sulfur-oxidizing bacteria were enumerated and isolated from a steady-state anaerobic master culture reactor (MCR) operated for over six months under a semi-continuous mode and nitrate-limiting conditions using thiosulfate as an electron donor. Most are Gram-negative bacteria, which have sizes up to 12 m. Strains AD1 and AD2 were isolated from the plate count agar (PCA), and strains BD1 and BD2 from the solid thiosulfate/nitrate medium. Based on the morphological, physiological, FAME and 16S rDNA sequence analyses, the two dominant strains, AD1 and AD2, were identified as Paracoccus denitrificans and Paracoccus versutus (formerly Thiobacillus versutus), respectively. From the physiological results, glucose was assimilated by both strains AD1 and AD2. Heterotrophic growth of strains AD1 and AD2 could be a more efficient way of obtaining a greater amount of biomass for use as an inoculum. Even though facultative autotrophic bacteria grow under heterotrophic conditions, autotrophic denitrification would not be reduced.

• Molecules and Cells
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• v.44 no.1
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• pp.26-37
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• 2021

Human papillomaviruses (HPVs) cause cellular hyperproliferation-associated abnormalities including cervical cancer. The HPV genome encodes two major viral oncoproteins, E6 and E7, which recruit various host proteins by direct interaction for proteasomal degradation. Recently, we reported the structure of HPV18 E7 conserved region 3 (CR3) bound to the protein tyrosine phosphatase (PTP) domain of PTPN14, a well-defined tumor suppressor, and found that this intermolecular interaction plays a key role in E7-driven transformation and tumorigenesis. In this study, we carried out a molecular analysis of the interaction between CR3 of HPV18 E7 and the PTP domain of PTPN21, a PTP protein that shares high sequence homology with PTPN14 but is putatively oncogenic rather than tumor-suppressive. Through the combined use of biochemical tools, we verified that HPV18 E7 and PTPN21 form a 2:2 complex, with a dissociation constant of 5 nM and a nearly identical binding manner with the HPV18 E7 and PTPN14 complex. Nevertheless, despite the structural similarities, the biological consequences of the E7 interaction were found to differ between the two PTP proteins. Unlike PTPN14, PTPN21 did not appear to be subjected to proteasomal degradation in HPV18-positive HeLa cervical cancer cells. Moreover, knockdown of PTPN21 led to retardation of the migration/invasion of HeLa cells and HPV18 E7-expressing HaCaT keratinocytes, which reflects its protumor activity. In conclusion, the associations of the viral oncoprotein E7 with PTPN14 and PTPN21 are similar at the molecular level but play different physiological roles.

• Journal of Veterinary Science
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• v.23 no.6
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• pp.12.01-12.10
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• 2022

Background: Staphylococcus aureus and Streptococcus agalactiae are the main cause of clinical mastitis in dairy cattle in Argentina, whereas coagulase-negative staphylococci (CNS) and environmental streptococci are the main cause of subclinical mastitis. Bacteria isolated from infected animals show increasing antimicrobial resistance. Objectives: This study aims to determine the antimicrobial resistance of staphylococci and streptococci isolated from milk with mastitis, and to genotypically characterize the methicillin-resistant (MR) staphylococci. Methods: Isolation was performed on blood agar and identification was based on biochemical reactions. Antimicrobial susceptibility was according to the Clinical and Laboratory Standards Institute guidelines. The antimicrobial resistance genes, SCCmec type and spa type were detected by the polymerase chain reaction method. Results: We isolated a total of 185 staphylococci and 28 streptococci from 148 milk samples. Among the staphylococcal isolates, 154 were identified as CNS and 31 as S. aureus. Among the 154 CNS, 24.6% (n = 38) were resistant to penicillin, 14.9% (n = 23) to erythromycin, 17.5% (n = 27) to clindamycin, 6.5% (n = 10) to cefoxitin and oxacillin. Among the S. aureus isolates, 16.1% (n = 5) were resistant to penicillin, 3.2% (n = 1) to cefoxitin and oxacillin (MRSA). Six MR isolates (5 CNS and 1 MRSA) were positive to the mecA gene, and presented the SCCmec IVa. The MRSA strain presented the sequence type 83 and the spa type 002. Among the 28 streptococcal isolates, 14.3% (n = 4) were resistant to penicillin, 10.7% (n = 3) to erythromycin and 14.3% (n = 4) to clindamycin. Conclusions: The present findings of this study indicate a development of antimicrobial resistance in main bacteria isolated from cows with mastitis in Argentina.

• Animal Bioscience
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• v.35 no.3
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• pp.399-409
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• 2022

Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC₅₀) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC₅₀ levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.

• Molecules and Cells
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• v.45 no.12
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• pp.886-895
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• 2022

Malignant rhabdoid tumor (MRT) is a highly aggressive pediatric malignancy with no effective therapy. Therefore, it is necessary to identify a target for the development of novel molecule-targeting therapeutic agents. In this study, we report the importance of the runt-related transcription factor 1 (RUNX1) and RUNX1-Baculoviral IAP (inhibitor of apoptosis) Repeat-Containing 5 (BIRC5/survivin) axis in the proliferation of MRT cells, as it can be used as an ideal target for anti-tumor strategies. The mechanism of this reaction can be explained by the interaction of RUNX1 with the RUNX1-binding DNA sequence located in the survivin promoter and its positive regulation. Specific knockdown of RUNX1 led to decreased expression of survivin, which subsequently suppressed the proliferation of MRT cells in vitro and in vivo. We also found that our novel RUNX inhibitor, Chb-M, which switches off RUNX1 using alkylating agent-conjugated pyrrole-imidazole polyamides designed to specifically bind to consensus RUNX-binding sequences (5'-TGTGGT-3'), inhibited survivin expression in vivo. Taken together, we identified a novel interaction between RUNX1 and survivin in MRT. Therefore the negative regulation of RUNX1 activity may be a novel strategy for MRT treatment.

• Journal of Marine Life Science
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• v.1 no.2
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• pp.79-87
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• 2016

Heavy metals such as cadmium (Cd) are highly toxic to aquatic organisms and human, even at trace concentration. Herein we investigated the effect of Cd on the gene expression of ATP-binding cassette (ABC) transporters and glutathione S-transferase (GST) in marine ciliate Euplotes crassus. Seven ABC transporters and one GST genes were partially cloned and sequences, and thereafter, transcriptional modulation of these genes after exposure to Cd for 8 h was investigated using quantitative real time RT- PCR (qRT-PCR). As results, sequence analysis and phylogenetic study revealed that E. crassus ABCs are likely typical ABC transports, in particular, B/C family, and GST gene may be similar to GST theta isoform. A significant increase in the expression of ABCs, except for ABCB21 was observed in a concentration dependent manner after exposure to Cd (0.1 and 0.5 mg/l) for 8 h. The GST mRNA level was the highest at 0.5 mg/l Cd and then reduced until control level. These findings suggest that ABCs and GST may be involved in a protective mechanism against Cd-mediated toxicity in E. crassus.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.457-466
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• 2024

Cellobiose dehydrogenases (CDHs) are a group of enzymes belonging to the hemoflavoenzyme group, which are mostly found in fungi. They play an important role in the production of acid sugar. In this research, CDH annotated from the actinobacterium Cellulomonas palmilytica EW123 (CpCDH) was cloned and characterized. The CpCDH exhibited a domain architecture resembling class-I CDH found in Basidiomycota. The cytochrome c and flavin-containing dehydrogenase domains in CpCDH showed an extra-long evolutionary distance compared to fungal CDH. The amino acid sequence of CpCDH revealed conservative catalytic amino acids and a distinct flavin adenine dinucleotide region specific to CDH, setting it apart from closely related sequences. The physicochemical properties of CpCDH displayed optimal pH conditions similar to those of CDHs but differed in terms of optimal temperature. The CpCDH displayed excellent enzymatic activity at low temperatures (below 30℃), unlike other CDHs. Moreover, CpCDH showed the highest substrate specificity for disaccharides such as cellobiose and lactose, which contain a glucose molecule at the non-reducing end. The catalytic efficiency of CpCDH for cellobiose and lactose were 2.05 × 10⁵ and 9.06 × 10⁴ (M^-1 s^-1), respectively. The result from the Fourier-transform infrared spectroscopy (FT-IR) spectra confirmed the presence of cellobionic and lactobionic acids as the oxidative products of CpCDH. This study establishes CpCDH as a novel and attractive bacterial CDH, representing the first report of its kind in the Cellulomonas genus.

• The Journal of the Petrological Society of Korea
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• v.27 no.3
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• pp.139-151
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• 2018

The Songgang-ri area, Cheongsong-gun, which is located in the Sobaeksan province of Yeongnam Massif near the southwestern boundary of Yeongyang subbasin of Gyeongsang Basin, consists of age unknown metamorphic rocks (banded gneiss, granitic gneiss, limesilicates) and age unknown igneous rock (granite gneiss) which intrudes them. This paper researched the geological structures of the Songgang-ri area from the geometric and kinematic features and the developing sequence of multi-deformed rock structures in the geological outcrops exposed about 170 m along the riverside of Yongjeoncheon in the eastern part of Songgang village, Songgang-ri. In the Songgang-ri geological outcrops are recognized three times (Fn, Fn+1, Fn+2) of folding, three times (Dk-I, Dk-II, Dk-III) intrusion of acidic dykes, one time of faulting, which are different in deformation and intrusion timing each other. These geological structures are at least formed by five times (Dn, Dn+1, Dn+2, Dn+3, Dn+4) of deformation. The Dn deformation is recognized by Fn fold which axial surface is parallel to the regional foliation. The Dn+1 intruded the (E)NE trending Dk-I dyke in the earlier phase and formed the NW trending Fn+1 fold in the later phase under compression of (E)NE-(W)SW direction. There are tight, isoclinal, intrafolial folds, boudinage, ${\sigma}$- or ${\delta}$-type boudins, asymmetric fold, C' shear band as the major deformed rock structures. The Dn+2 intruded the (N)NW trending Dk-II dyke in the earlier phase and formed NE trending Fn+2 fold in the later phase under compression of (N)NW-(S)SE direction. There are open fold and folded boudinage as those. The Dn+2 intruded the Dk-III dyke which cuts the Dk-I and Dk-II dykes and the axial surface of Fn+2 fold. The Dn+3 formed the left-handed reverse oblique-slip fault of NNE trend in which hanging wall moves into the SSE direction. Considering in that such five times of deformation recognized in the Songgang-ri geological outcrops are closely connected to the distribution and geological structure of the constituents in the more regional area as well as Songgang-ri area, the research result is expected to play a great data in clarifying and understanding the geological structure and its development process of the surrounding and boundary constituents of the Yeongnam Massif and Gyeongsang Basin.

• Tuberculosis and Respiratory Diseases
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• v.62 no.5
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• pp.406-416
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• 2007

Background: Single nucleotide polymorphisms (SNPs), which consist of a substitution of a single nucleotide pair, are the most abundant form of genetic variations occurring with a frequency of approximately 1 per 1000 base pairs. SNPs by themselves do not cause disease but can predispose humans to disease, modify the extent or severity of the disease or influence the drug response and treatment efficacy. Single nucleotide polymorphisms (SNPs), particularly those within the regulatory regions of the genes often influence the expression levels and can modify the disease. Studies examining the associations between SNP and the disease outcome have provided valuable insight into the disease etiology and potential therapeutic intervention. Traditionally, the genotyping of SNPs has been carried out using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP), which is a low throughput technique not amenable for use in large-scale SNP studies. Recently, TaqMan real-time PCR chemistry was adapted for use in allelic discrimination assays. This study validated the accuracy and utility of real-time PCR technology for SNPs genotyping Methods: The SNPs in promoter sequence (-37 and -524) of lung cancer suppressor gene, RRM1 (ribonucleotide reductase M1 subunit) with the genomic DNA samples of 89 subjects were genotyped using both real-time PCR and PCR-RFLP. Results: The discordance rates were 2.2% (2 mismatches) in -37 and 16.3% (15 mismatches) in -524. Auto-direct sequencing of all the mismatched samples(17 cases) were in accord with the genotypes read by real-time PCR. In addition, 138 genomic DNAs were genotyped using real-time PCR in a duplicate manner (two separated assays). Ninety-eight percent of the samples showed concordance between the two assays. Conclusion: Real-time PCR allelic discrimination assays are amenable to high-throughput genotyping and overcome many of the problematic features associated with PCR-RFLP.

• The Korean Journal of Quaternary Research
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• v.16 no.2
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• pp.9-21
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• 2002

In Korea, many open-air upper palaeolithic sites are located at the river valley, particularly exposed in gently rotting terrain along the river course. They are situated at an altitude less trail 30 m above present river bottom, and covered with the blankets of slope deposits of several meters in thickness. The purpose of this research is to eluridate depositional and vegetational environment of the alluvial upper palaeolithic Jangheung-ri sites on the basis of analytical properties of grain size population, chronology, palynology, soil chemistry and clay mineralogy and magnetic susceptibility of the Jangheung-ri Quaternary formations. The lithostratograpy of Jangheung-ri sit is subdivided into 3 layers based on the depositional sequence and radiocarbon ages. From bottom to top, they are composed of slope deposits with lower paleosol layers, young fluvial sand and gravel with backswamp organic muds, and upper paleosol layers. The upper paleosol was formed under rather dry climatic condition between each flooding period. Dessication cracks were prevalent in the soil solum which was filled with secondarily minuted fragments due to pedogenetic process. The soil structure shows typical braided-typed cracks in the root part of cracking texture, and more diversified pattern of crackings downward. The young fluvial sand gravel were formed by rather perennial streams after LGM. The main part of organic muds was particularly formed after 15Ka. Local backswamp were flourished with organic muds and graded suspension materials in the flooding muds were intermittently accumulated in the organic muds until ca. 11Ka. This episode was associated with migration of Nam River toward present course. Organic muds were formed in backswamp or local pond. Abies/Picea-Betula with Ranunculaceae, Compositae, Cyperaceae were prevalent. This period is characterized with B$\Phi$lling, Older Dryas, Allerod, and Younger Dryas (MIS-1). Stone artefacts were found in the lower paleosol layers formed as old as 18Ka-22Ka. Based on the artefacts and landscape settings of the Jangheung-ri site, it is presumed that settlement grounds of old people were buried by frequent floodings of old Nam River, the river-beds of which were heavily fluctuated laterally and river-bed erosions were activated from south to north in Jangheung-ri site until the terminal of LGM9ca 17Ka).