• Biomolecules & Therapeutics
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• v.22 no.2
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• pp.114-121
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• 2014

Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.

• The Journal of Natural Sciences
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• v.14 no.1
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• pp.7-24
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• 2004

We found new type of thiol peroxidase, fused with GRX.(TPx-GRX) The TPx-GRX exists in pathogenic bacteria including -. This protein was homogeneously purified from the E.coli recombinant overexpressing TPx-GRX. In the presence of a thiol-containing electron donor such as DTT, the purified TPx-GRX has potent the antioxidant to prevent the inactivation of GS by the MCO system, which is comprised of DTT, $Fe^{3+}$, and $O^2$. The antioxidant activity is much higher that other thiol peroxidase. The investigate the peroxidase activity of TPx-GRX, we directly measured the peroxidase activity of TPx-GRX toward peroxides in terms of the removal of peroxides in the presence of GSH. This result demonstrates that the peroxidase activity of TPx-GRX. These taken together results suggest that TPx-GRX is a new member of thiol peroxidase. These observations also suggest that in the pathogenic bacteria, TPx-GRX plays an important antioxidative role as a multiple array defence mechanism against oxidative stress.

• ALGAE
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• v.35 no.4
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• pp.389-404
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• 2020

Red algal fertilization is unusual and offers a different model to the mechanism of intracellular transport of nuclei and polyspermy blocking. A female carpogonium (egg) undergoes plasmogamy with many spermatia (sperm) simultaneously at the receptive structure, trichogyne, which often contains numerous male nuclei. The pattern of selective transport of a male nucleus to the female nucleus, located in the cell body of the carpogonium, remain largely unknown. We tracked the movement of spermatial nuclei and cell organelles in the trichogyne after plasmogamy using time-lapse videography and fluorescent probes. The fertilization process of Bostrychia moritziana is composed of five distinctive stages: 1) gamete-gamete binding; 2) mitosis in the attached spermatia; 3) formation of a fertilization channel; 4) migration of spermatial nuclei into the trichogyne; and 5) cutting off of the trichogyne cytoplasm from the rest of the cell after karyogamy. Our results showed that actin microfilaments were involved in the above steps of fertilization, microtubules are involved only in spermatial mitosis. Time-lapse videography showed that the first ("primary") nucleus which entered to trichogyne moved quickly to the base of carpogonium and fused with the female nucleus. The transport of the primary male nucleus to the egg nucleus was complete before its second nucleus migrated into the trichogyne. Male nuclei from other spermatia stopped directional movement soon after the first one entered the carpogonial base and oscillated near where they entered trichogyne. The cytoplasm of the trichogyne was cut off at a narrow neck connecting the trichogyne and carpogonial base after gamete nuclear fusion but gamete binding and plasmogamy continued on the trichogyne. Spermatial organelles, including mitochondria, entered the trichogyne together with the nuclei but did not show any directional movement and remained close to where they entered. These results suggest that polyspermy blocking in B. moritziana is achieved by the selective and rapid transport of the first nucleus entered trichogyne and the rupture of the trichogyne after gamete karyogamy.

• Clinical and Experimental Pediatrics
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• v.49 no.4
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• pp.401-409
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• 2006

Purpose : Human metapneumovirus(hMPV) is a respiratory viral pathogen that causes a wide spectrum of illnesses, ranging from asymptomatic infection to severe bronchiolitis. The virus has been identified world widely, but so far it has not been published in Korea. Methods : We obtained clinical samples by nasopharyngeal aspiration from 218 children hospitalized due to acute lower respiratory tract infections at Soonchunhyang University Hospital in Cheonan from October, 2004 to April, 2005. We designed specific primers from conserved region of fusion glycoprotein of hMPV. Total RNA was extracted and RT-PCR was performed, and single specific 423 bp product was obtained. The PCR product was confirmed to be fusion glycoprotein RNA by sequencing. Results : We detected hMPV in 15(6.9 percent) of the 218 hospitalized children. The infected children comprised nine boys and six girls; their mean age was 2.8 years(5 mo-12 yrs) and they were diagnosed with pneumonia(60 percent), bronchiolitis(33.3 percent), croup(6.6 percent). The number of cases of detected hMPV in Korea increased dramatically during the period from March to May 2005. Conclusion : hMPV is circulating in Korean children and is associated with respiratory tract infection. Additional studies are required to define the epidemiology and the extent of diseases in the general population caused by hMPV.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1636-1643
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• 2012

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

• Journal of Korean Neurosurgical Society
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• v.65 no.6
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• pp.834-840
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• 2022

Objective : C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR), and white blood cell (WBC) count are inflammatory markers used to evaluate postoperative infections. Although these markers are non-specific, understanding their normal kinetics after surgery may be helpful in the early detection of postoperative infections. To compliment the recent trend of reducing the duration of antibiotic use, this retrospective study investigated the inflammatory markers of patients who had received antibiotics within 24 hours after surgery according to the Health Insurance Review & Assessment Service guidelines and compared them with those of patients who had received antibiotics for 5 days, which was proven to be non-infectious. Methods : We enrolled 74 patients, divided into two groups. Patients underwent posterior lumbar interbody fusion (PLIF) at a single institution between 2019 and 2020. Group A included 37 patients who received antibiotics within 24 hours after the PLIF procedure, and group B comprised 37 patients who had used antibiotics for 5 days. A 1 : 1 nearest-neighbor propensity-matched analysis was used. The clinical variables included age, sex, medical history, body mass index, estimated blood loss, and operation time. Laboratory data included CRP, ESR, and WBC, which were measured preoperatively and on postoperative days (POD) 1, 3, 5, and 7. Results : CRP dynamics tended to decrease after peaking on POD 3, with a similar trend in both groups. The average CRP level in group B was slightly higher than that in group A; however, the difference was not statistically significant. Multiple linear regression analysis revealed operation time, number of fused levels, and estimated blood loss as significant predictors of a greater CRP peak value (r²=0.473, p<0.001) in patients. No trend (a tendency to decrease from the peak value) could be determined for ESR and WBC count on POD 7. Conclusion : Although slight differences were observed in numerical values and kinetics, sequential changes in inflammatory markers according to the duration of antibiotic administration showed similar patterns. Knowledge of CRP kinetics allows the assessment of the degree of difference between the clinical and expected values.

• Journal of the Korea Society of Computer and Information
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• v.28 no.1
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• pp.49-54
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• 2023

The Global Positioning System (GPS) was developed for military purposes and developed as it is today by opening civilian signals (GPS L1 frequency C/A signals). The current satellite orbits the earth about twice a day to measure the position, and receives more than 3 satellite signals (initially, 4 to calculate even the time error). The three-dimensional position of the ground receiver is determined using the data from the radio wave departure time to the radio wave Time of Arrival(TOA) of the received satellite signal through trilateration. In the case of navigation using GPS in recent years, a location error of 5 to 10 m usually occurs, and quite a lot of areas, such as apartments, indoors, tunnels, factory areas, and mountainous areas, exist as blind spots or neutralized areas outside the error range of GPS. Therefore, in order to acquire one's own location information in an area where GPS satellite signal reception is impossible, another method should be proposed. In this study, IMU(Inertial Measurement Unit) combined with an acceleration and gyro sensor and a geomagnetic sensor were used to design a system to enable location recognition even in terrain where GPS signal reception is impossible. A method to track the current position by calculating the instantaneous velocity value using a 9-DOF IMU and a geomagnetic sensor was studied, and its feasibility was verified through production and experimentation.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.14
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• pp.6073-6080
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• 2015

Background: Melanoma differentiation-associated gene-7 (MDA-7)/interleukin-24 (IL-24), a unique tumor suppressor gene, has killing activity in a broad spectrum of cancer cells. Herein, plasmids producing mda-7 proteins fused to different RGD peptides (full RGD4C and shortened RGD, tRGD) were evaluated for apoptosis induction with a hepatocellular carcinoma cell line, Hep-G2. The study aim was to improve the apoptosis potency of mda-7 by tethering to RGD peptides. Materials and Methods: Three plasmids including mda-7, mda-7-RGD and mda-7-tRGD genes beside a control vector were transfected into Hep-G2 cells. After 72 hours incubation, cell viability was evaluated by MTT assay. In addition, the rate of apoptosis was analyzed by flow cytometry using PI/annexin staining. To detect early events in apoptosis, 18 hours after transfection, expression of the BAX gene was quantified by real time PCR. Modeling of proteins was also performed to extrapolate possible consequences of RGD modification on their structures and subsequent attachment to receptors. Results and Conclusions: In MTT assays, while all mda-7 forms showed measurable inhibition of proliferation, unmodified mda-7 protein exhibited most significant effect compared to control plasmid (P<0.001). Again, flow cytometry analysis showed a significant apoptosis induction by simple mda-7 gene but not for those RGD-fused mda-7 proteins. These findings were also supported by expression analysis of BAX gene (P<0.001). Protein modelling analysis revealed that tethering RGD at the end of IL-24/Mda7 disrupt attachment to cognate receptor, IL-20R1/IL-20R2. In conclusion, fusion of RGD4C and shortened RGD peptides to carboxyl terminal of mda7, not only reduce apoptosis property in vitro but also disrupt receptor attachment as demonstrated by protein modelling.

• 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.

• The Journal of Korean Society for School & Community Health Education
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• v.22 no.4
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• pp.49-60
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• 2021

Objectives: Laboratories have various latent physical, chemical, biological, and ergonomical factors according to the diversification and fusion of research and development activities. This study aims to investigate the chemical exposure concentrations of college laboratories and evaluate their health risks, and use them as basic data to promote the health of college students. Methods: The sampling and analysis of harmful chemicals in the air in laboratories were performed using Method 1500 of the U.S. National Institute for Occupational Safety and Health (NIOSH)의 Method 1500. The harmful chemicals in the laboratories were divided into carcinogenic and non-carcinogenic chemicals. Risk assessment was performed using the cancer risk (CR) for carcinogenic chemicals and using the hazard index (HI) for non-carcinogenic chemicals. Results: The harmful chemicals in college laboratories consisted of acetone, diethyl ether, methylene chloride, n-hexane, ethyl acetate, chloroform, tetrahydrofuran, toluene, and xylenes. They showed the highest concentrations in laboratories A (acetone 0.001~2.34ppm), B (chloroform 0.95~6.35ppm), C (diethyl ether 0.08~8.68ppm), and D (acetone 0.07~14.96ppm). The risk assessment result for non-carcinogenic chemicals showed that the HI of methylene chloride was 2.052 for men and 2.333 for women, the HI of N-hexane was 4.442 for men and 5.05 for women. Thus, the HI values were higher than 1. The risk of carcinogenic chemicals is determined by an excess cancer risk (ECR) value of 1.0×10^-5, which means that one in 100,000 people has a cancer risk. The ECRs of chloroform exceeded 1.0×10^-5 for both men and women, indicating the possibility of cancer risk. Conclusion: College laboratories showed the possibility of non-carcinogenic health risks for methylene chloride, n-hexane, tetrahydrofuran (THF), toluene, and xylenes, and carcinogenic health risks for chloroform, methylene chloride. However, this study used the maximum values of measurements to determine the worst case, and assumed that the subjects were exposed to the corresponding concentrations continuously for 8 hours per day for 300 days per year. In consideration of the nature of laboratory environment in which people are intermittently exposed, rather than continuously, to the chemicals, the results of this study has an element of overestimation.