• Asian Pacific Journal of Cancer Prevention
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• v.14 no.7
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• pp.4209-4214
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• 2013

Background: Breast cancer is among the five most common cancers and ranks first among cancers diagnosed in Iranian women. Screening and treatment of this disease with molecular methods, especially regarding high incidences at early age and advanced stage, is essential. Several genes with altered expression have been identified by cDNA microarray studies in breast cancer, with the Bcl-2 gene indicated as a likely candidate. In this study, we studied Bcl-2 gene expression levels in parallel tumor and non-tumor breast tissues. Materials and Methods: Forty samples including 21 tumor, 16 non tumor (marginal) and 3 benign breast tissues which were all pathologically diagnosed, were subjected to RNA extraction and polyA RT-PCR with the expression level of Bcl-2 quantified using real-time PCR. Results: There is higher expression levels of the Bcl-2 gene in tumor samples compared with marginal samples, but not attaining significance(p>0.05). Bcl-2 expression in 14 (66.7%) of the cases of tumor samples and 9 (56.3%) cases of the marginal samples were positive. Comparison of the expression of the Bcl-2 gene in histological grade showed that a high expression of Bcl-2 was associated with a high histological grade (p<0.41). Conclusions: Our data suggests that dysregulated Bcl-2 gene expression is potentially involved in the pathogenesis of breast cancer. Using gene expression analysis may significantly improve our ability for screening cancer patients and will prove a powerful tool in the diagnosis and prognostic evaluation of the disease whilst aiding the cooperative group trials in the Bcl-2 based therapy project.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.1
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• pp.17-27
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• 2001

A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

• Journal of dental hygiene science
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• v.15 no.3
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• pp.377-382
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• 2015

The purpose of this study was to evaluate the validity and reliability of plaque scoring system using new Qraycam (All in One Bio, Korea) device which enables plaque score without tooth disclosing. This study measured Quigley-Hein index and plaque control record by both Qraycam and disclosing agent on 64 elderly people and checked degree of congruence between the two methods. Reliability was evaluated with the mean of measured values, kappa index and intraclass correlation coefficient statistical analysis. The analysis of the plaque scores showed a high agreement between the measured values according to the method of measurement and the measured part. The mean of plaque index of anterior labial were not significantly different according to measurement method. The kappa index was higher by Qraycam and tooth disclosing method of plaque index. Therefore, it was verified that Qraycam has sufficient reliability as screening tool for plaque scoring system.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.408-420
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• 2014

Yeast surface-displayed antibody libraries provide an efficient and quantitative screening resource for given antigens, but suffer from typically modest library sizes owing to low yeast transformation efficiency. Yeast mating is an attractive method for overcoming the limit of yeast transformation to construct a large, combinatorial antibody library, but the optimal conditions have not been reported. Here, we report a large synthetic human Fab (antigen binding fragment) yeast surface-displayed library generated by stepwise optimization of yeast mating conditions. We first constructed HC (heavy chain) and LC (light chain) libraries, where all of the six CDRs (complementarity-determining regions) of the variable domains were diversified mimicking the human germline antibody repertoires by degenerate codons, onto single frameworks of VH3-23 and $V{\kappa}1$-16 germline sequences, in two haploid cells of opposite mating types. Yeast mating conditions were optimized in the order of cell density, media pH, and cell growth phase, yielding a mating efficiency of ~58% between the two haploid cells carrying HC and LC libraries. We constructed two combinatorial Fab libraries with CDR-H3 of 9 or 11 residues in length with colony diversities of more than $10^9$ by one round of yeast mating between the two haploid HC and LC libraries, with modest diversity sizes of ${\sim}10^7$. The synthetic human Fab yeast-displayed libraries exhibited relative amino acid compositions in each position of the six CDRs that were very similar to those of the designed repertoires, suggesting that they are a promising source for human Fab antibody screening.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.167-178
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• 2020

Lipase-producing fungi have been isolated from environments containing lipids. The non-dairy creamer industrial waste has a high amount of lipids so it is a potential source for the isolation of lipase-producing fungi. However, the study of fungi that secrete lipase from this industrial waste has not been reported. The purpose of this study was to obtain lipase-producing filamentous fungi from non-dairy creamer industrial waste. Mineral salt and potato dextrose agar were used as media for the isolation process. The qualitative screening was conducted using phenol red agar medium and the quantitative screening using broth medium containing glucose and olive oil. Isolates producing the highest amounts of lipase were identified with molecular methods. We found that 5 out of 19 isolated filamentous fungi are lipase producers. Further analysis showed that isolate Ms.11 produced the highest amount of lipase compared to others. Based on ITS sequence Ms.11 was identified as Aspergillus aculeatus. The lipase activity in medium containing 1% glucose + 1% olive oil at pH 7.0 and 30℃ after 96 and 120 h of incubation was 5.13 ± 0.30 U/ml and 5.22 ± 0.59 U/ml, respectively. The optimum lipase activity was found at pH 7.0, 30℃ and using methanol or ethanol in the reaction tube. Lipase was more stable at 20-30℃ and maintained 85% of its activity. It was concluded that isolate Ms.11 is a potential source of lipase that catalyzes transesterification reactions. Further studies are required to optimize lipase production to make the strain suitable for industry purposes.

• Journal of the Korea Convergence Society
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• v.10 no.3
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• pp.23-30
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• 2019

The purpose of this study is to find out the structure of the substance that affects antidiabetic using the candidate drug information for diabetes treatment. A quantitative structure activity relationship model based on machine learning method was constructed and major molecular descriptors were determined for each experimental data variables from coefficient values using a partial least squares algorithm. The results of the analysis of the molecular access system fingerprint data reflecting the candidate drug structure information were higher than those of the in vitro data analysis in terms of goodness-of-fit, and the major molecular expression factors affecting the antidiabetic effect were also variously derived. If the proposed method is applied to the new drug development environment, it is possible to reduce the cost for conducting candidate screening experiment and to shorten the search time for new drug development.

• Journal of Environmental Health Sciences
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• v.48 no.5
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• pp.272-281
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• 2022

Background: The constant consumption of chemical products owing to expanding industrialization has led to an increase in public interest in chemical substances. As the production and disposal processes for these chemical products cause environmental problems, regional information on the hazard level of chemical substances is required considering their effects on humans and in order to ensure environmental safety. Objectives: This study aimed to identify hazard contribution and spatiotemporal characteristics by region and chemical by calculating a hazard-based result score using pollutant release and transfer register (PRTR) data. Methods: This study calculated the chemical discharge and hazard-based result score from the Risk-Screening Environmental Indicators (RSEI) model, analyzed their spatiotemporal patterns, and identified hotspot areas where chemical discharges and high hazard-based scores were concentrated. The amount of chemical discharge and hazard-based risk scores for 250 cities and counties across South Korea were calculated using PRTR data from 2011 to 2018. Results: The chemical discharge (high densities in Incheon, Daegu, and Busan) and hazard-based result scores (high densities in Incheon, Chungcheongnam-do, and some areas of Gyeongsangnam-do Province) showed varying spatial patterns. The chemical discharge (A, B) and hazard-based result score (C, D) hotspots were identified. Additionally, identification of the hazard-based result scores revealed differences in the type of chemicals contributing to the discharge. Ethylbenzene accounted for ≥80% of the discharged chemicals in the discharge hotspots, while chromium accounted for >90% of the discharged chemicals in the hazard-based result score hotspots. Conclusions: The RSEI hazard-based result score is a quantitative indicator that considers the degree of impact on human health as a toxicity-weighted value. It can be used for the management of industries discharging chemical substances as well as local environmental health management.

• Journal of Genetic Medicine
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• v.7 no.1
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• pp.59-66
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• 2010

Purpose: Quantitative fluorescent polymerase chain reaction (QF-PCR) allows for the rapid prenatal diagnosis of common aneuploidies. The main advantages of this assay are its low cost, speed, and automation, allowing for large-scale application. However, despite these advantages, it is not a routine method for prenatal aneuploidy screening in Korea. Our objective in the present study was to validate the performance of QF-PCR using short tandem repeat (STR) markers in a Korean population as a means for rapid prenatal diagnosis. Material and Methods: A QF-PCR assay using an Elucigene kit (Gen-Probe, Abingdon, UK), containing 20 STR markers located on chromosomes 13, 18, 21, X and Y, was performed on 847 amniotic fluid (AF) samples for prenatal aneuploidy screening referred for prenatal aneuploidy screening from 2007 to 2009. The results were then compared to those obtained using conventional cytogenetic analysis. To evaluate the informativity of STR markers, the heterozygosity index of each marker was determined in all the samples. Results: Three autosomes (13, 18, and 21) and X and Y chromosome aneuploidies were detected in 19 cases (2.2%, 19/847) after QF-PCR analysis of the 847 AF samples. Their results are identical to those of conventional cytogenetic analysis, with 100% positive predictive value. However, after cytogenetic analysis, 7 cases (0.8%, 7/847) were found to have 5 balanced and 2 unbalanced chromosomal abnormalities that were not detected by QF-PCR. The STR markers had a slightly low heterozygosity index (average: 0.76) compared to those reported in Caucasians (average: 0.80). Submicroscopic duplication of D13S634 marker, which might be a unique finding in Koreans, was detected in 1.4% (12/847) of the samples in the present study. Conclusion: A QF-PCR assay for prenatal aneuploidy screening was validated in our institution and proved to be efficient and reliable. However, we suggest that each laboratory must perform an independent validation test for each STR marker in order to develop interpretation guidelines of the results and must integrate QF-PCR into the routine cytogenetic laboratory workflow.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.12
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• pp.1665-1671
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• 2013

Real-time quantitative PCR (qRT-PCR) is one of the important methods for investigating the changes in mRNA expression levels in cells and tissues. Selection of the proper reference genes is very important when calibrating the results of real-time quantitative PCR. Studies on the selection of reference genes in goat tissues are limited, despite the economic importance of their meat and dairy products. We used real-time quantitative PCR to detect the expression levels of eight reference gene candidates (18S, TBP, HMBS, YWHAZ, ACTB, HPRT1, GAPDH and EEF1A2) in ten tissues types sourced from Boer goats. The optimal reference gene combination was selected according to the results determined by geNorm, NormFinder and Bestkeeper software packages. The analyses showed that tissue is an important variability factor in genes expression stability. When all tissues were considered, 18S, TBP and HMBS is the optimal reference combination for calibrating quantitative PCR analysis of gene expression from goat tissues. Dividing data set by tissues, ACTB was the most stable in stomach, small intestine and ovary, 18S in heart and spleen, HMBS in uterus and lung, TBP in liver, HPRT1 in kidney and GAPDH in muscle. Overall, this study provided valuable information about the goat reference genes that can be used in order to perform a proper normalisation when relative quantification by qRT-PCR studies is undertaken.

• Journal of Marine Life Science
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• v.8 no.1
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• pp.68-77
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• 2023

In this study, we investigated the quantitative distribution of microplastics in the surface seawater at 8 points near the Taean Peninsula using fluorescence staining. The study revealed a detection range of microplastics from 0 to 360.5 particles/l, with an average of 149.7 ± 46.0 particles/l. When classifying the microplastics by size, it was found that particles smaller than 50 ㎛ were dominant, although there were differences at Site 3. Moreover, it was not possible to identify clear correlations when comparing the number of microplastics based on collection area and particle size. Various physical and chemical factors, including plastic material, dynamic ocean conditions (such as currents, wind, waves, tides), geological characteristics (topography, slope), sediment materials including coastal organisms, human activities (fishing, development, tourism), and weather conditions (floods, rainfall), affect the behavior of microplastics. Therefore, future efforts should focus on standardizing quantitative analysis methods and conducting fundamental research on microplastic monitoring, including the analysis of environmental factors.