• Title/Summary/Keyword: R-gene

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Nuclear SSU and Plastid rbcL Genes and Ultrastructure of Mallomonas caudata (Synurophyceae) from Korea (한국산 Mallomonas caudata (Synurophyceae)의 미세구조, 핵 SSU 그리고 색소체 rbcL 유전자)

  • Kim, Han-Soon;Shin, Woong-Ghi;Boo, Sung-Min
    • Korean Journal of Ecology and Environment
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    • v.40 no.3
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    • pp.387-394
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    • 2007
  • Despite geographic barriers such as oceans, many freshwater algal species inhabit different continents of the world. A unicellular freshwater alga, Mallomonas caudeata, commonly occurring in Asia, Europe, and America of the northern Hemisphere, is closely related to human life such as monitoring blooms and defecting changes in climates. In order to demonstrate its occurrence in Korea and to infer its phylogeny, we sequenced nuclear SSU and plastid rbcL genes from isolates collected in six different reservoirs. We have also investigated transmission electron microscopy of the Korean isolates. SSU sequences of the species from Korea and USA were almost identical, having pair-wise divergences of 0.06% in SSU and 0.45% in rbcL. Both gene trees revealed that the species was clearly separated from other species of the genus, while the genus was not monophyletic. Rhizoplasts are composed of microfibrils organised in striated rootlets attached to the multilayered plate of basal bodies and arranged on the surface of the nucleus at their distal ends. The rhizoplast constitutes a basal body-nucleus connector similar to that of typical Synurophyceas. The results that Mallomonas was not supported by both SSU and rbeL data sets require a further study with additional taxon sampling.

Comparison of Conventional Culture Method and Real-time PCR for Detection of Yersinia enterocolitica in Sausage and Vegetable Salad (소시지와 야채 샐러드에서 Yersinia enterocolitica 검출을 위한 배지법과 real-time PCR법의 비교)

  • Kim, Yun-Gyeong;Chon, Jung-Whan;Lee, Jae-Hoon;Kwak, Hyo-Sun;Hwang, In-Gyun;Seo, Kun-Ho
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.133-136
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    • 2013
  • The purpose of this study was to compare a conventional culture method and real-time PCR for the detection of Yersinia enterocolitica (Y. enterocolitica) in sausage and in vegetable salad. Food samples inoculated with Y. enterocolitica were enriched in peptone-sorbitol bile-broth, and swabs were then streaked onto cefsulodin-irgasan-novobiocin agar. Biochemical tests for suspected colonies were performed with an API 20E strip. In parallel, real-time PCR was performed, targeting the 16S rRNA gene using 1 mL of enrichment broth. In sausage, the number of positive samples detected by culture method (49 out of 60) was similar (p>0.05) with that of real-time PCR (50 out of 60). However, the number of positive samples of real-time PCR (26 out of 60) was significantly higher (p<0.05) than that of the conventional culture method (6 out of 60) in vegetable salad. Real-time PCR could be an effective screening tool for detecting Y. enterocolitica, particularly in food samples with high levels of background flora, such as a vegetable salad.

Characterization of Bacillus licheniformis SCK A08 with Antagonistic Property Against Bacillus cereus and Degrading Capacity of Biogenic Amines (Bacillus cereus에 대한 길항적 저해 작용과 biogenic amines 분해 능력을 지닌 Bacillus licheniformis SCK A08 균의 특성)

  • Lee, Eon Sil;Kim, Yong Sang;Ryu, Myeong Seon;Jeong, Do Yeon;Uhm, Tai Boong;Cho, Sung Ho
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.40-46
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    • 2014
  • We have screened Bacillus strains suitable for the fermentation of soybean products with respect to the control of Bacillus cereus and the reduction of biogenic amines. Of 26 isolates, a strain named as the SCK A08 carried antimicrobial activity against B. cereus and Staphylococcus aureus, major food poisoning species in soybean products. PCR analysis revealed that the SCK A08 strain did not contain genes for Bacillus cereus toxins including nonhemolytic enterotoxin, hemolytic enterotoxin, cytotoxin K, cereulide and certrax. The SCK A08 strain could degrade histamine, tyramine, putrescine, and cadaverine by 67.41%, 76.59%, 57.32%, and 50.69%, respectively, during fermentation in cooked soybeans containing 0.5% (w/w) of each biogenic amine. The morphological and biochemical properties and phylogenetic relationships based on 16S rRNA gene sequences indicated that the isolate was most closely related to Bacillus licheniformis. Use of the strain SCK A08 would be a potential measure to overcome two hygienic problems that were frequently faced during manufacture of traditionally fermented soybean products.

Whole genome MBD-seq and RRBS analyses reveal that hypermethylation of gastrointestinal hormone receptors is associated with gastric carcinogenesis

  • Kim, Hee-Jin;Kang, Tae-Wook;Haam, Keeok;Kim, Mirang;Kim, Seon-Kyu;Kim, Seon-Young;Lee, Sang-Il;Song, Kyu-Sang;Jeong, Hyun-Yong;Kim, Yong Sung
    • Experimental and Molecular Medicine
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    • v.50 no.12
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    • pp.1.1-1.14
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    • 2018
  • DNA methylation is a regulatory mechanism in epigenetics that is frequently altered during human carcinogenesis. To detect critical methylation events associated with gastric cancer (GC), we compared three DNA methylomes from gastric mucosa (GM), intestinal metaplasia (IM), and gastric tumor (GT) cells that were microscopically dissected from an intestinal-type early gastric cancer (EGC) using methylated DNA binding domain sequencing (MBD-seq) and reduced representation bisulfite sequencing (RRBS) analysis. In this study, we focused on differentially methylated promoters (DMPs) that could be directly associated with gene expression. We detected 2,761 and 677 DMPs between the GT and GM by MBD-seq and RRBS, respectively, and for a total of 3,035 DMPs. Then, 514 (17%) of all DMPs were detected in the IM genome, which is a precancer of GC, supporting that some DMPs might represent an early event in gastric carcinogenesis. A pathway analysis of all DMPs demonstrated that 59 G protein-coupled receptor (GPCR) genes linked to the hypermethylated DMPs were significantly enriched in a neuroactive ligand-receptor interaction pathway. Furthermore, among the 59 GPCRs, six GI hormone receptor genes (NPY1R, PPYR1, PTGDR, PTGER2, PTGER3, and SSTR2) that play an inhibitory role in the secretion of gastrin or gastric acid were selected and validated as potential biomarkers for the diagnosis or prognosis of GC patients in two cohorts. These data suggest that the loss of function of gastrointestinal (GI) hormone receptors by promoter methylation may lead to gastric carcinogenesis because gastrin and gastric acid have been known to play a role in cell differentiation and carcinogenesis in the GI tract.

Effects of Glucagon-Like Peptide-2-Expressing Saccharomyces cerevisiae Not Different from Empty Vector

  • Zhong, Xi;Liang, Guopeng;Cao, Lili;Qiao, Qi;Hu, Zhi;Fu, Min;Bo, Hong;Wu, Qin;Liang, Guanlin;Zhang, Zhongwei;Zhou, Lin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1644-1655
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    • 2019
  • Saccharomyces cerevisiae (S. cerevisiae) and glucagon-like peptide-2 (GLP-2) have been employed to improve the intestinal development of weaned animals. The goal of this study was to determine whether either exogenous S. cerevisiae or GLP-2 elicits major effects on fecal microbiotas and cytokine responses in weaned piglets. Ninety-six piglets weaned at 26 days were assigned to one of four groups: 1) Basal diet (Control), 2) empty vector-harboring S. cerevisiae (EV-SC), 3) GLP-2-expressing S. cerevisiae (GLP2-SC), and 4) recombinant human GLP-2 (rh-GLP2). At the start of the post-weaning period (day 0), and at day 28, fecal samples were collected to assess the bacterial communities via sequencing the V1-V2 region of the 16S-rRNA gene, and piglets' blood was also sampled to measure cytokine responses (i.e., IL-$1{\beta}$, TNF-${\alpha}$, and IFN-${\gamma}$). This study revealed that, on the one hand, although S. cerevisiae supplementation did not significantly alter the growth of weaned piglets, it induced increases in the relative abundances of two core genera (Ruminococcaceae_norank and Erysipelotrichaceae_norank) and decreases in the relative abundances of two other core genera (Lachnospiraceae_norank and Clostridiale_norank) and cytokine levels (IL-$1{\beta}$ and TNF-${\alpha}$) (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). On the other hand, GLP-2 supplementation had no significant influence on fecal bacterial communities and cytokine levels, but it produced better body weight and average daily gain (p < 0.05, Control vs EV-SC; p < 0.05, rh-GLP2 vs GLP2-SC). Therefore, altered fecal microbiotas and cytokine response effects in weaned piglets were due to S. cerevisiae rather than GLP-2.

Deacidification Effect of Campbell Early Must through Carbonic-Maceration Treatment: Isolation and Properties of the Bacteria Associated with Deacidification (Carbonic Maceration처리에 의한 Campbell Early 발효액의 감산 효과: 감산 관련 미생물의 분리 및 특성)

  • Chang, Eun-Ha;Jeong, Seok-Tae;Jeong, Sung-Min;Lim, Byung-Sun;Noh, Jung-Ho;Park, Kyo-Sun;Park, Seo-Jun;Choi, Jong-Uck
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.973-979
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    • 2011
  • The grape cultivar Campbell Early has high levels of malic acid as well as tartaric acid. The high concentration of total acid in the Campbell Early wine is a critical aspect of the wine's sensory characteristics. To prevent the deterioration of the wine's quality, which is caused by the strong sour taste derived from the raw material in wine making, the deacidification factor was investigated via carbonic maceration under different temperature conditions, especially in the presence or absence of malolactic bacteria. Based on the results of the presence test of the malolactic bacteria during carbonic-maceration treatment, Lactobacillus brevis, Lactobacillus plantarum, and Streptococcus thermophilus were characterized morphologically and were identified via biochemical tests and 16S-rRNA-gene-sequencing analysis. The isolated strains were found not to consume malic acid and to produce lactic acid. Moreover, these strains were consumed as soluble solids. The isolated strains are popularly known as lactic-acid bacteria and should have produced lactic acid from glucose. The Oenococcus oeni of the malolactic bacteria was not isolated. These results showed that the isolated strains are not deacidified during carbonic-maceration treatment.

Microbial Diversity and Community Analysis in Lettuce or Cucumber Cultivated Greenhouse Soil in Korea (상추 및 오이 시설재배 토양의 미생물 다양성 분석)

  • Kim, Byung-Yong;Weon, Hang-Yeon;Park, In-Cheol;Lee, Sang-Yeob;Kim, Wan-Gyu;Song, Jae-Kyeong
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.6
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    • pp.1169-1175
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    • 2011
  • The soil chemical properties, microbial community structures and biochemical properties of lettuce or cucumber-cultivated greenhouse soil samples were analyzed to assess soil health and characterize microbial distribution in 8 locations in Korea. Although most of chemical properties were within the soil management guidelines, the available phosphate, and the contents of exchangeable potassium and calcium were higher than those of recommended levels. In the culture-dependent analysis, 841 bacterial strains were isolated from the greenhouse soils and were identified at the genus level by 16S rRNA gene sequences analysis. The dominant bacterial genera were Bacillus (35.7%), Microbacterium (9.3%), Arthrobacter (5.7%) and Lysobacter (5.1%). The abundance of pseudomonads was highly variable depending on the soil samples. In the culture-independent analysis, soil microbial community was investigated by using phospholipid fatty acid (PLFA) method. Principal component analysis (PCA) showed that a specific grouping for microbial community structure in the greenhouse soils was not observed based on cultivated crops and investigated sites. The results revealed that the greenhouses soils examined are relatively sound managed in terms of soil chemical contents and microbial properties.

Enhancing Butyrate Production, Ruminal Fermentation and Microbial Population through Supplementation with Clostridium saccharobutylicum

  • Miguel, Michelle A.;Lee, Sung Sill;Mamuad, Lovelia L.;Choi, Yeon Jae;Jeong, Chang Dae;Son, Arang;Cho, Kwang Keun;Kim, Eun Tae;Kim, Sang Bum;Lee, Sang Suk
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1083-1095
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    • 2019
  • Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on $NH_3-N$ at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, $NH_3-N$ and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with $10^6CFU/ml$ C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

Walnut phenolic extracts reduce telomere length and telomerase activity in a colon cancer stem cell model

  • Shin, Phil-Kyung;Zoh, Yoonchae;Choi, Jina;Kim, Myung-Sunny;Kim, Yuri;Choi, Sang-Woon
    • Nutrition Research and Practice
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    • v.13 no.1
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    • pp.58-63
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    • 2019
  • BACKGROUND/OBJECTIVES: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. MATERIALS AND METHODS: $CD133^+CD44^+$ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and $40{\mu}g/mL$ for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA ($ddC_t$). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA ($dC_t$). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. RESULTS: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner ($5.16{\pm}0.13$ at $0{\mu}g/mL$, $4.79{\pm}0.12$ at $10{\mu}g/mL$, $3.24{\pm}0.08$ at $20{\mu}g/mL$ and $3.99{\pm}0.09$ at $40{\mu}g/mL$; P = 0.0276). Telomerase activities concurrently decreased with telomere length ($1.47{\pm}0.04$, $1.09{\pm}0.01$, $0.76{\pm}0.08$, and $0.88{\pm}0.06$; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. CONCLUSION: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.

Clinical Characteristics of Korean Patients with Lung Cancer Who Have Programmed Death-Ligand 1 Expression

  • Park, Ha-Young;Oh, In-Jae;Kho, Bo Gun;Kim, Tae-Ok;Shin, Hong-Joon;Park, Cheol Kyu;Kwon, Yong-Soo;Kim, Yu-Il;Lim, Sung-Chul;Kim, Young-Chul;Choi, Yoo-Duk
    • Tuberculosis and Respiratory Diseases
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    • v.82 no.3
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    • pp.227-233
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    • 2019
  • Background: Programmed death-ligand 1 (PD-L1), a transmembrane protein, binds to the programmed death-1 (PD-1) receptor, and anti-PD-1 therapy enables immune responses against tumors. This study aimed to assess clinical characteristics of PD-L1 expression using immunohistochemistry among Korean patients with lung cancer. Methods: We retrospectively reviewed the data of patients with pathologically proven lung cancer from a single institution. PD-L1 expression determined by Tumor Proportion Score (TPS) was detected using 22C3 pharmDx (Agilent Technologies) and SP263 (Ventana Medical Systems) assays. Results: From July 2016 to July 2017, 267 patients were enrolled. The main histologic type was adenocarcinoma (69.3%). Most participants were smokers (67.4%) and had clinical stage IV disease (60.7%). In total, 116 (42%) and 58 (21%) patients had TPS ${\geq}1%$ and ${\geq}50%$, respectively. The patients were significantly older in TPS ${\geq}1%$ group than in TPS <1% group ($64.83{\pm}9.38years$ vs. $61.73{\pm}10.78years$, p=0.014), not in TPS ${\geq}50%$ cutoff value ($64.69{\pm}9.39$ vs. $62.36{\pm}10.51$, p=0.178). Regarding histologic grade, higher proportions of poorly differentiated tumor were observed in the TPS ${\geq}1%$ (40.8% vs. 25.8%, p=0.020) and TPS ${\geq}50%$ groups (53.2% vs. 27.2%, p=0.004). Among 34 patients examined with 22C3 and SP263 assays, 27 had positive results in both assays, with a cutoff of TPS ${\geq}1%$ (r=0.826; 95% confidence interval, 0.736-0.916). Conclusion: PD-L1 expression, defined as TPS ${\geq}1%$, was related to older age and poorly differentiated histology. There was a similar distribution of PD-L1 expression in both 22C3 and SP263 results.