• Title/Summary/Keyword: molecular distribution

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Induction of Interleukin-8 Expression in Synovial Cell by Hepatitis C Virus Core Protein (활막 세포에서 HCV Core 단백에 의한 Interleukin-8 발현 유도)

  • Wang, Jin-Sang;Her, Won-Hee;Kim, So-Yeon;Yoon, Seung-Kew
    • IMMUNE NETWORK
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    • v.6 no.1
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    • pp.20-26
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    • 2006
  • Background: Rheumatoid arthritis (RA) is a chronic and systemic inflammatory disease that is characterized by invasive synovial hyperplasia, leading to progressive joint destruction. Recent studies have described that RA is caused by virus, bacteria or outside material. Approximately 2 to 20% of RA cases arc reported to be associated with infected hepatitis C virus (HCV). However, the mechanisms underlying virus-induced RA are still unknown. Moreover, few molecular studies have addressed the inflammatory aspects of HCV-associated autoimmune RA. In this study, we aimed to determine whe ther or not another HCV core protein transactivates the IL-8 gene expression, prototypic chemokine, in synovial cell. Methods: To establish the HCV core expressing stable synovial cell line, pCI-neo-core, a plasmid encoding HCV core protein, were transfected to HIG-82 cell line that is an established cell line from rabbit periaricular soft tissue. We examined the morphological changes and cell cycle distribution of HIG-82 cells with expression of HCV core protein by inverted microscopy and flow cytometry analysis, respectively. Also, we determined the mRNA levels of Interleukin (IL)-6 and IL-8 related to the inflammation by RT-PCR and then analyzed regulation of IL-8 expression by the NF-${\kappa}B$ pathway. Results: Our study showed no significant differences in morphology and cell cycle between HIG-82 control cell line and HIG-82 expressing HCV core protein. However, expression of HCV core protein induces the IL-8 mRNA expression in HIG-82 core cells via activated NF-${\kappa}B$ pathway. Conclusion: These results suggest that HCV core protein can lead to enhanced IL-8 expression. Such a proinflammatory role may contribute to the etiologic pathogenesis in RA patients with HCV infection.

Vitamin D Sufficiency: How should it be defined and what are its functional indicators?

  • Hollis Bruce W.
    • Proceedings of the Korean Nutrition Society Conference
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    • 2004.11a
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    • pp.22-33
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    • 2004
  • It has been more than three decades since the first assay assessing circulating 25(OH)D in human subjects was performed. That publication as well as several that followed it defined 'normal' nutritional vitamin D status in human populations. Recently, the wisdom by which 'normal' circulating 25(OH)D levels in human subjects were assigned in the past has come under question. It appears that sampling human subjects, who appear to be free from disease, and assessing 'normal' circulating 25(OH)D levels by plotting a Gaussian distribution is grossly inaccurate. There are many reasons why this method is inaccurate, including race, lifestyle habits, sunscreen usage, age, latitude, and inappropriately low dietary recommendations for vitamin D. For instance, a 400IU/day. AI for vitamin D is insignificant when one considers that a 10-15 minute whole body exposure to peak summer sun will generate and release up to 20,000 IU vitamin $D_3$ into the circulation. Recent studies, which orally administered up to 10,000 IU/day vitamin $D_3$ to human subjects for several months, have successfully elevated circulating 25(OH)D levels to those observed in individuals from sun-rich environments. Further, we are now able to accurately assess sufficient circulating 25(OH)D levels utilizing specific biomarkers instead of guessing what an adequate level is. These biomarkers include intact parathyroid hormone (PTH), calcium absorption, bone mineral density (BMD), insulin resistance and pancreatic beta cell function. Using the data from these biomarkers, vitamin D deficiency should be defined as circulating levels of $25(OH)D{\leq}30ng/mL$. In certain cases, such as pregnancy and lactation, significantly higher circulating 25(OH)D levels would almost certainly be beneficial to both the mother and recipient fetus/infant.

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Cloning and characterization of a novel gene with alternative splicing in murine mesenchymal stem cell line C3H/10T1/2 by gene trap screening

  • Wang, Mingke;Sun, Huiqin;Jiang, Fan;Han, Jing;Ye, Feng;Wang, Tao;Su, Yongping;Zou, Zhongmin
    • BMB Reports
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    • v.43 no.12
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    • pp.789-794
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    • 2010
  • A novel gene, designated mgt-6, containing four splicing variants, was isolated from a gene trap clone library of C3H/10T1/2 cells transfected with retroviral promoterless gene-trap vector, ROSAFARY. The transcript variants were differentially expressed in murine tissues and cell lines and differentially responded to diverse stimuli including TGF-${\beta}1$ and mitogen-activated protein kinase (MAPK) inhibitors. The mgt-6 gene encoded a protein of 37 or 11 amino acid residuals with cytoplasmic distribution. However, when C3H/10T1/2 cells were treated with 5-azacytidine, the protein translocated into cell nucleus as indicated by fused LacZ or C-terminally tagged EGFP. Our preliminary results suggest that further study on the role of mgt-6 gene in cell transformation and differentiation may be of significance.

Macrolepiota in Korea: New Records and a New Species

  • Cho, Hae Jin;Lee, Hyun;Park, Myung Soo;Kim, Changmu;Wisitrassameewong, Komsit;Lupala, Abel;Park, Ki Hyeong;Kim, Min Ji;Fong, Jonathan J.;Lim, Young Woon
    • Mycobiology
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    • v.47 no.4
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    • pp.368-377
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    • 2019
  • The genus Macrolepiota (Agaricales, Basidiomycota) is easy to recognize at the genus level because of big, fleshy basidiocarps with squamules covering the pileus; a single or double annulus; and big, thick-walled basidiospores with a germ pore. However, morphological identification is often unreliable in Macrolepiota due to similar morphological features among species. Due to the uncertainty of previous morphological identification in the genus Macrolepiota, it is necessary to re-examine Korean Macrolepiota using molecular data. We reexamined 34 Macrolepiota specimens collected from 2012 to 2018 in Korea using a reverse taxonomic approach, whereby species identification was first done based on the internal transcribed spacer (ITS) region analysis, followed by morphological confirmation. We identified the presence of four species: M. detersa, M. mastoidea, M. procera, and M. umbonata sp. nov. Two species (M. detersa and M. mastoidea) were previously unrecorded from Korea and M. umbonata is a new species. Detailed descriptions of all four species and taxonomic key are provided in this study. Macrolepiota procera and M. umbonata are distributed through the country, but M. detersa and M. mastoidea are distributed only in limited areas. According to our results, the combination of ITS locus and morphology proved to be a robust approach to evaluate the taxonomic status of Macrolepiota species in Korea. Additional surveys are needed to verify the species diversity and clarify their geographic distribution.

The Effect of Camphorsulfonic Acid in TEMPO-Mediated Bulk and Dispersion Polymerization of Styrene

  • Oh Sejin;Kim Gijung;Ko Narae;Shim Sang Eun;Choe Soonja
    • Macromolecular Research
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    • v.13 no.3
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    • pp.187-193
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    • 2005
  • The TEMPO-mediated living free-radical bulk and dispersion polymerization of styrene in the presence of camphorsulfonic acid (CSA) are investigated. In the absence of TEMPO and CSA in the bulk polymerization, a conversion of $93\%$ is achieved within 6 hr of polymerization. When only TEMPO is involved in this polymerization, the pseudo-living free-radical polymerization is well achieved, however, the polymerization rate becomes quite slow. This retardation of the polymerization rate is solved by the addition of a low concentration of CSA. In the TEMPO-mediated dispersion polymerization in the presence of CSA, similar trends in the conversion, kinetics, and PDI are observed as those observed in the case of bulk polymerization. When only TEMPO is used in the dispersion polymerization, the resulting particle size becomes quite broad, due to the prolonged polymerization time. However, when a 1.0 molar ratio of CSA to TEMPO is added to the TEMPO-mediated dispersion polymerization, fairly mono-disperse PS microspheres having an average size of 5.83 $\mu$m and a CV of 3.4$\%$ are successfully obtained, due to the narrow molecular weight distribution of the intermediate oligomers and shortening of the polymerization time. This result indicates that the addition of CSA to the TEMPO-mediated bulk and the use of dispersion polymerization not only shortens the polymerization time, but also greatly improves the uniformity of the microspheres.

Genomic DNA Extracted from Ancient Antarctic Glacier Ice for Molecular Analyses on the Indigenous Microbial Communities

  • Lee, Sang-Hoon;Bidle, Kay;Falkowski, Paul;Marchant, David
    • Ocean and Polar Research
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    • v.27 no.2
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    • pp.205-214
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    • 2005
  • From ancient Antarctic glacier ice, we extracted total genomic DNA that was suitable for prokaryotic 16S rDNA gene cloning and sequencing, and bacterial artificial chromosome (BAC) library and end-sequencing. The ice samples were from the Dry Valley region. Age dating by $^{40}Ar/^{39}Ar$ analysis on the volcanic ashes deposited in situ indicated the ice samples are minimum 100,000-300,000 yr (sample DLE) and 8 million years (sample EME) old. Further assay proved the ice survived freeze-thaw cycles or other re-working processes. EME, which was from a small lobe of the basal Taylor glacier, is the oldest known ice on Earth. Microorganisms, preserved frozen in glacier ice and isolated from the rest of the world over a geological time scale, can provide valuable data or insight for the diversity, distribution, survival strategy, and evolutionary relationships to the extant relatives. From the 16S gene cloning study, we detected no PCR amplicons with Archaea-specific primers, however we found many phylotypes belonging to Bacteria divisions, such as Actinobacteria, Acidobacteria, Proteobacteria $({\alpha},\;{\beta},\;and\;{\gamma})$, Firmicutes, and Cytophaga-Flavobacterium-Bacteroid$. BAC cloning and sequencing revealed protein codings highly identical to phenylacetic acid degradation protein paaA, chromosome segregation ATPases, or cold shock protein B of present day bacteria. Throughput sequencing of the BAC clones is underway. Viable and culturable cells were recovered from the DLE sample, and characterized by their 16S rDNA sequences. Further investigation on the survivorship and functional genes from the past should help unveil the evolution of life on Earth, or elsewhere, if any.

Analysis of Spiramycin in Fish Using High Performance Liquid Chromatography (HPLC를 이용한 어류 중의 Spiramycin 분석)

  • Lee Tae-Seek;Lee Hee-Jung;Jo Mi-Ra;Byun Han-Seok;Son Kwang-Tae;Park Mi-Jung;Yi Young-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.2
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    • pp.78-84
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    • 2006
  • A high performance liquid chromatography assay method for spiramycin in fish muscle was developed. The developed method was evaluated and validated by monitoring spiramycin in olive flounder (Paralichthys olivaceus), black rock fish (Sebastes schlegeli) and in live conger eel (Anguilla japonica) in fish farms and distribution centers. Using the developed method, the recovery rate was up to 82.4-88.8%, which was higher than that of conventional methods (77.6-87.1%). In particular, the proposed sample treatment protocol was suitable for use with fish samples to remove low molecular weight materials and pigments that could interfere an accurate analysis. The prepared stock solution was very stable, and it remained chemically stable for 5 weeks at $4^{\circ}C$. The performance limit of the developed method for spiramycin acid in fish muscle was 0.05 ppm. One hundred thirty-four fish samples including olive flounder, black rock fish and live conger eel were analyzed to evaluate the overall efficiency of the modified method and to monitor the actual condition of spiramycin usage in fish farms. Olive flounder and black rock fish were collected from fish farms in coastal areas of Korea, and live conger eels were purchased from a fish market in the Busan area from September 2001 to March 2002. According to the overall performance of the developed method, it was considered suitable for the monitoring of spiramycin in fish muscle. The suggested method of analysis for spiramycin in fish muscle is registered in the Korean Official Methods of Food Analysis and is currently tieing utilized for fishery products inspection by the Korea Food and Drug Administration and the National Fisheries Products Quality Inspection Service.

Molecular Diversity of Bacterial Communities from Subseafloor Rock Samples in a Deep-Water Production Basin in Brazil

  • Von Der Weid, Irene;Korenblum, Elisa;Jurelevicius, Diogo;Rosado, Alexandre Soares;Dino, Rodolfo;Sebastian, Gina Vasquez;Seldin, Lucy
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.5-14
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    • 2008
  • The deep subseafloor rock in oil reservoirs represents a unique environment in which a high oil contamination and a very low biomass can be observed. Sampling this environment has been a challenge owing to the techniques used for drilling and coring. In this study, the facilities developed by the Brazilian oil company PETROBRAS for accessing deep subsurface oil reservoirs were used to obtain rock samples at 2,822-2,828 m below the ocean floor surface from a virgin field located in the Atlantic Ocean, Rio de Janeiro. To address the bacterial diversity of these rock samples, PCR amplicons were obtained using the DNA from four core sections and universal primers for 16S rRNA and for APS reductase (aps) genes. Clone libraries were generated from these PCR fragments and 87 clones were sequenced. The phylogenetic analyses of the 16S rDNA clone libraries showed a wide distribution of types in the domain bacteria in the four core samples, and the majority of the clones were identified as belonging to Betaproteobacteria. The sulfate-reducing bacteria community could only be amplified by PCR in one sample, and all clones were identified as belonging to Gammaproteobacteria. For the first time, the bacterial community was assessed in such a deep subsurface environment.

Seasonal and Spatial Diversity of Picocyanobacteria Community in the Great Mazurian Lakes Derived from DGGE Analyses of 16S rDNA and cpcBA-IGS Markers

  • Jasser, Iwona;Krolicka, Adriana;Jakubiec, Katarzyna;Chrost, Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.739-749
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    • 2013
  • The seasonal and spatial diversity of picocyanobacteria (Pcy) in lakes of the Great Mazurian Lakes (GLM) system was examined by DGGE analysis of molecular markers derived from the 16S-23S internal transcribed spacer (ITS) of the ribosomal operon and the phycocyanin operon (cpcBA-IGS). The study of nine lakes, ranging from mesotrophy to hypereutrophy, demonstrated seasonal variance of Pcy. The richness and Shannon diversity index calculated on the basis of both markers were higher in spring and lower in early and late summer. No statistically significant relationships were found between the markers and trophic status of the studied lakes or Pcy abundance. There were, however, statistically significant relationships between the diversity indices and sampling time. The analysis pointed to a different distribution of the two markers. The ITS marker exhibited more unique sequences in time and space, whereas a greater role for common and ubiquitous sequences was indicated by the cpcBA-IGS data. Examination of the Pcy community structure demonstrated that communities were grouped in highly similar clusters according to sampling season/time rather than to the trophic status of the lake. Our results suggest that time is more important than trophic status in shaping the diversity and structure of Pcy communities. The seasonal changes in picocyanobacteria and differences in diversity and community structures are discussed in the context of well-established ecological hypotheses: the PEG model, intermediate disturbance hypothesis (IDH), and horizontal gene transfer (HGT).

Isolation of High-molecular-weight-compound degrading microorganisms and sulfate reducing Bacteria involved in Composting Process (퇴비화 과정에 관여하는 생체 고분자 분해 미생물 및 황산 환원균의 분리)

  • Lee, Seong-Taek;Lee, Jae-Jeong;Na, Hyun-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.2 no.2
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    • pp.31-37
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    • 1994
  • For a microbiological study of composting process, screening and assay method for biopolymer degrading enzymes and microorganisms were developed and for the study of the possibility of composting in anaerobic state, distribution of sulfate reducing bacteria which plays a final role in anaerobic degradation was investigated. Substrates used for the development of assay methods for biopolymer degradation are ${\beta}-glucan$, xylan, dextran, CMC(carboxy methly cellulose), casein, and collagen. These substrates were made insoluble by a cross-linking agent and linked with dye to make chromogenic substrates. ${\beta}-glucan$ and xylan substrates could substitute congo-red method for screening of polymer degrading microorganisms without damaging the colonies. Sulfate reducing bacteria contained in the sample sludge showed preference to lactic acid, propionic acid, butyric acid and formic acid and could use acetic acid and valeric acid.

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