• Title/Summary/Keyword: DNA density

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Pathological Factors Affecting DNA Quality in BRAF, EGFR, and KRAS Gene Molecular Tests (BRAF, EGFR, KRAS 유전자 분자병리검사에서 DNA 품질에 영향을 미치는 병리학적인 인자에 관한 연구)

  • Yun, Hyon-Goo;Kim, Bo-Ra;Lee, Joo-Mi;Song, Eun-Ha;Kim, Dong-Hoon
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.4
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    • pp.381-388
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    • 2020
  • The quality control of pathological specimens is important for accurate molecular pathology testing. This study evaluated that specimen factors affecting the DNA quality during tissue processing and sample types for BRAF, EGFR, and KRAS mutations tests. One thousand seven hundred and seventy-two molecular pathology tests were investigated for the factors influencing the DNA quality, such as sample type, formalin fixation time, and reexamination status. Cytology samples stored in a saline solution had better DNA quality than commercial cytology preservation. Tissue samples fixed in formalin within 24 hours had better DNA quality than the samples fixed over 24 hours. Between the types of samples, fresh tissue samples and tissue samples with a high tumor cell density had relatively better DNA quality than the formalin-fixed paraffin-embedded (FFPE) tissues and cytology specimens. Of real-time PCR, the non-PNA Ct value increased proportionally with samples held for longer than 24 hours in formalin, and that the formalin-fixed time affects the sample DNA quality. In conclusion, the appropriate tumor cellularity and 10% neutral formalin fixation time are the most important factors for maintaining the DNA quality. These factors should be managed properly for an accurate pathological molecular test to ensure optimal DNA quality.

Monitoring the presence of wild boar and land mammals using environmental DNA metabarcoding - Case study in Yangpyeong-gun, Gyeonggi-do - (환경 DNA 메타바코딩을 활용한 멧돼지 및 육상 포유류 출현 모니터링 - 경기도 양평군 일대를 중심으로 -)

  • Kim, Yong-Hwan;Han, Youn-Ha;Park, Ji-Yun;Kim, Ho Gul;Cho, Soo-Hyun;Song, Young-Keun
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.24 no.6
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    • pp.133-144
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    • 2021
  • This study aims to estimate location of land mammals habitat by analyzing spatial data and investigate how to apply environmental DNA monitoring methodology to lotic system in Yangpyeong-gun, Gyeonggi-do. Environmental DNA sampling points are selected through spatial analysis with QGIS open source program by overlaying Kernel density of wild boar(Sus scrofa), elevation, slope and land-cover map, and 81 samples are collected. After 240 mL of water was filtered in each sample, metabarcoding technique using MiMammal universal primer was applied in order to get a whole list of mammal species whose DNA particles contained in filtered water. 8 and 22 samples showed DNA of wild boar and water deer, respectively. DNA of raccoon dog, Eurasian otter, and Siberian weasel are also detected through metabarcoding analysis. This study is valuable that conducted in outdoor lotic system. The study suggests a new wildlife monitoring methodology integrating overlayed geographic data and environmental DNA.

Nanoscale Fabrication of Biomolecular Layer and Its Application to Biodevices

  • Park, Jeong-Woo;Nam, Yun-Suk;Masamichi Fujihira
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.76-85
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    • 2004
  • Biodevices composed of biomolecular layer have been developed in various fields such as medical diagnosis, pharmaceutical screening, electronic device, photonic device, environmental pollution detection device, and etc. The biomolecules such as protein, DNA and pigment, and cells have been used to construct the biodevices such as biomolecular diode, biostorage device, bioelectroluminescence device, protein chip, DNA chip, and cell chip. Substantial interest has focused upon thin film fabrication or the formation of biomaterials mono- or multi-layers on the solid surfaces to construct the biodevices. Based on the development of nanotechnology, nanoscale fabrication technology for biofilm has been emerged and applied to biodevices due to the various advantages such as high density immobilization and orientation control of immoblized biomolecules. This review described the nanoscale fabrication of biomolecular film and its application to bioelectronic devices and biochips.

High Frequency Electroporation-transformation of Coryneform Bacteria Grown in the Medium with Penicillin-G (Penicillin-G 첨가 배지에서 배양한 코리네형 세균의 전기장 충격법에 의한 고효율 형질전환)

  • 노갑수;김성준
    • KSBB Journal
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    • v.6 no.3
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    • pp.223-230
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    • 1991
  • Using the shuttle vector pECCGl between Escherichia coli and Corynebacterium glutamicum and C. glutamicum strain JS231 grown in the medium supplemented with penicillin-G, which inhibits the formation of cross-links in the peptidoglycan of bacterial cell wall, various parameters involved in electroporation system including resistance, electric field strength, capacitance, DNA concentration, and cell density were investigated independently and optimized for the high efficiency transformation of coryneform bacteria. Using cells grown with 0.3U/ml of penicillin-G and harvested at A600 of 0.7-0.8, transformation efficiencies of 107-l08 transformants/$\mu\textrm{g}$ of DNA with Corynebcctertum glutamicum strain JS231 and wild type ATCC13032 were achieved under conditions of 12.5kV/cm of electric field strength, 400 ohms of resistance, $25\mu$F of capacitance, 3$\times$108 cells per transformation(1.2$\times$1010 cells/ml) and 100ng of plasmid DNA per transformation.

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Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Effects of Cover Plants on Soil Microbial Community in a Organic Pear Orchard

  • Oh, Young-Ju;Sohn, Soo-In;Song, Yang-Ik;Kang, Seok-Boem;Choi, Jin-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.1
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    • pp.28-35
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    • 2014
  • Due to recent interest of the consumers on safe farm products and the government's political support for eco-friendly agriculture, organic fruit production has been growing continuously. This research was conducted in order to study the effect of cover plants on soil microbial community on cover plants and establish an organic fruit cultivation method through choosing optimal cover plant. As a result of investigating soil microbial population density, the bacterial density in soil showed an increasing trend in June compared to April, and there was a decreasing trend in bacterial density of the soil in August compared to June. The density of actinomycetes in soil increased around 1.6 times in June compared to April when the soil was covered with hairy vetch. The increase of filamentous fungus in crimson clover group was 6.1 times higher in June compared to April and in hairy vetch group, the increase was 4.9 times higher in June compared to April. As a result of analyzing DNA extracted from the soil categorized by different types of cover plants using DGGE method, soil collected from April had higher number of bands detected from different locations according to different types of cover plants. Diversity of the bands from the soil collected from August showed higher range of reduction. As a result of analyzing soil microbial community by different period and the types of cover plants using Pyrosequencing method, microbes were detected in the order of Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, and Firmicutes. Distribution rate of Firmicutes increased in the soil collected in August compared to June and this was shown in all types of cover plants by twice the amount.

Molecular Cloning of a cDNA Encoding Putative Apolipophorin from the Silkworm, Bombyx mori

  • Yun, Eun-Young;Goo, Tae-Won;Kim, Sung-Wan;Hwang, Jae-Sam;Park, Kwang-Ho;Kwon, O-Yu;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.2
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    • pp.145-149
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    • 2003
  • ApolipophorinIII (apoLp-III) is a protypical exchangeable apolipoprotein that is abundant in hemolymph of many insect species. Its function lies in the stabilization of low-density lipophorin particles (LDLp) crossing the hemocoel in phases of high energy consumption to deliver lipids from the fat body to the flight muscle cells. But, recent studies with naive Galleria mellonella-apoLp-III gave first indication of an unexpected role of that protein in insect immune activation. In this research, we cloned a cDNA encoding putative apoLp-III from the silkworm, Bombyx mori injected with E. coli and characterized its role. We constructed a cDNA library using whole bodies of B. mori larvae injected with E. coli, carried out the differential screening, and selected the up-regulated clones. Among these clones, we focused on a cDNA showing a high sequence similarity to the apolipophorinIII from other insects and analyzed the nucleotide and deduced amino acid sequences. The pupative B. mori Jam123 apoLp-III cDNA contained 1,131 bp encoding 186 amino acid residues. Phylogenetic analysis revealed that the nucleotide and amino acid sequences of the B. mori apoLp-III cDNA formed a highly inclusive subgroup with Bombycidae. But, it was interesting that B. mori Jam123 is closer to B. mandarina than B. mori P50 and B. mori N4. Northern blot analysis showed a signal in the fat body, posterior silkgland and midgut.

AN IN VITRO STUDY ON CELLULAR RESPONSE OF SEVERAL DENTURE BASE RESINS (수종 의치상 레진의 세포반응에 관한 연구)

  • Jun Chul-Oh;Vang Mong-Sook
    • The Journal of Korean Academy of Prosthodontics
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    • v.30 no.2
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    • pp.247-257
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    • 1992
  • The present study quantitates the in vitro cytotoxicity of a variety of denture base acrylic resins using cell culture techniques combined with image analysis to measure nuclear area and DNA contents. In this study, a comparison was made among direct curing, heat curing and microwave curing resins. The results obtained from this study were as follows : 1. Morphologically, cell process and nucleus became prominent but macroscopic difference according to the resins were nit observed. In addition, increased cellular density around the specimen were observed. 2. In DNA contents measurements, $S-G_2M$ phase cell was 15.47%, 14.58% in control and heat curing resin on 1st day and the others group $21.39\sim33.36%$ were measured. 3. Nuclear area and DNA contents were increased on 3rd day except DNA content of the microwave curing resin group. These results suggest that denture base acrylic resins stimulate gingival fibroblasts in vitro, especially stimulation of direct curing resin is larger and longer than the others.

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Electrochemical, Antifungal, Antibacterial and DNA Cleavage Studies of Some Co(II), Ni(II), Cu(II) and Zn(II)-Copolymer Complexes

  • Dhanaraj, C. Justin;Nair, M. Sivasankaran
    • Mycobiology
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    • v.36 no.4
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    • pp.260-265
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    • 2008
  • Cyclic voltammetric measurements were performed for Co(II), Ni(II), Cu(II) and Zn(II) complexes of 1 : 1 alternating copolymer, poly(3-nitrobenzylidene-1-naphthylamine-co-succinic anhydride) (L) and Ni(II) and Cu(II) complexes of 1 : 1 alternating copolymer, poly(3-nitrobenzylidene-1-naphthylamine-co-methacrylic acid) ($L^1$). The in vitro biological screening effects of the investigated compounds were tested against the fungal species including Aspergillus niger, Rhizopus stolonifer, Aspergillus flavus, Rhizoctonia bataicola and Candida albicans and bacterial species including Staphylococcus aureus, Escherichia coli, Klebsiella pneumaniae, Proteus vulgaris and Pseudomonas aeruginosa by well diffusion method. A comparative study of inhibition values of the copolymers and their complexes indicates that the complexes exhibit higher antimicrobial activity. Copper ions are proven to be essential for the growth-inhibitor effect. The extent of inhibition appeared to be strongly dependent on the initial cell density and on the growth medium. The nuclease activity of the above metal complexes were assessed by gel electrophoresis assay and the results show that the copper complexes can cleave pUC18 DNA effectively in presence of hydrogen peroxide compared to other metal complexes. The degradation experiments using Rhodamine B dye indicate that the hydroxyl radical species are involved in the DNA cleavage reactions.

Radical Scavenging Activities and Protective Effects against Oxidative Damage to DNA of Extracts from Medicinal Plants with known Osteoprotective Effects (골질 보호 작용이 있는 수종 생약 추출액의 래디칼 소거능 및 DNA 보호 효과)

  • Choi, Sung-Sook;Yim, Dong-Sool;Lee, Sook-Yeon
    • Korean Journal of Pharmacognosy
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    • v.40 no.2
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    • pp.143-149
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    • 2009
  • Among aging diseases, the most basic problem is a decrease in bone mineral density. Patients with osteoporosis are steadily increasing in the world. This study was investigated to show the relationship between osteoporotic effects and oxidative damage. Water extracts of 15 medicinal plants and ethanol extracts of 14 medicinal plants with known anti-osteoporotic effects, were tested for their radical scavenging activity using DPPH, ABTS, SRSA and FRAP assay. Water extract of Cornus officinalis, Rubus coreanus and ethanol extract of Rubus coreanus, Viscium album var. coloratum, Cimicifuga heracleifolia showed about 15-20 mg/g of total phenolic contents. Water extract of Cornus officinalis, Rubus coreanus and Epimedium koreanum showed high radical scavenging activity. Ethanol extract of Drynaria fortunei, Cornus officinalis, Rubus coreanus, Gentiana scabra and Astragalus membranaceus showed high radical scavenging activity. Water extract of Drynaria fortunei, Cornus officinalis, Nelumbo nucifera, Epimedium koreanum, and Gentiana scabra showed very strong protective effect against oxidative damage to DNA. These results suggest the correlation between anti-osteoporotic effects and antioxidative effects.