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Epigenetic insights into colorectal cancer: comprehensive genome-wide DNA methylation profiling of 294 patients in Korea

  • Soobok Joe;Jinyong Kim;Jin-Young Lee;Jongbum Jeon;Iksu Byeon;Sae-Won Han;Seung-Bum Ryoo;Kyu Joo Park;Sang-Hyun Song;Sheehyun Cho;Hyeran Shim;Hoang Bao Khanh Chu;Jisun Kang;Hong Seok Lee;DongWoo Kim;Young-Joon Kim;Tae-You Kim;Seon-Young Kim
    • BMB Reports
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    • v.56 no.10
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    • pp.563-568
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    • 2023
  • DNA methylation regulates gene expression and contributes to tumorigenesis in the early stages of cancer. In colorectal cancer (CRC), CpG island methylator phenotype (CIMP) is recognized as a distinct subset that is associated with specific molecular and clinical features. In this study, we investigated the genome-wide DNA methylation patterns among patients with CRC. The methylation data of 1 unmatched normal, 142 adjacent normal, and 294 tumor samples were analyzed. We identified 40,003 differentially methylated positions with 6,933 (79.8%) hypermethylated and 16,145 (51.6%) hypomethylated probes in the genic region. Hypermethylated probes were predominantly found in promoter-like regions, CpG islands, and N shore sites; hypomethylated probes were enriched in open-sea regions. CRC tumors were categorized into three CIMP subgroups, with 90 (30.6%) in the CIMP-high (CIMP-H), 115 (39.1%) in the CIMP-low (CIMP-L), and 89 (30.3%) in the non-CIMP group. The CIMP-H group was associated with microsatellite instability-high tumors, hypermethylation of MLH1, older age, and right-sided tumors. Our results showed that genome-wide methylation analyses classified patients with CRC into three subgroups according to CIMP levels, with clinical and molecular features consistent with previous data.

Changes in Internal and External Temperature and Microbiological Contamination depending on Consumer Behavior after Purchase of Fresh-Cut Produces (대형마트 신선편의식품 소비자의 구매 후 행동에 따른 식품 내·외부 온도 및 미생물학적 오염 변화)

  • Park, Hyun-Jin;Lee, Jeong-Eun;Kim, Sol-A;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.35 no.5
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    • pp.459-467
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    • 2020
  • In this study, we investigated the changes in both ambient temperature and microbial contamination of fresh convenience foods (FCFs) caused by the behavior of consumers after purchase. According to consumer survey results, it took 0.5 to 3 h put the purchased FCF in a home refrigerator or consume it. Only aerobic bacteria and Staphylococcus aureus (below maximum permitted limit) were detected in FCFs obtained from a local market. During storage of FCFs in a vehicle trunk for up to 3 h. the external and internal temperatures of FCFs were found to be 19 and 18.5℃ in spring, 44 and 42℃ in summer, 31.3 and 29.2℃ in autumn, and 17.6 and 16.8℃ in winter, respectively. Changes in contamination levels of aerobic bacteria on FCFs stored in a vehicle trunk for up to 3 hours are as follows: 2.72 → 3.41 log CFU/g in spring, 3.11 → 4.32 log CFU/g in summer, 3.08 → 3.81 log CFU/g in autumn, 2.71 → 3.36 log CFU/g in winter. S. aureus exceeding the tolerance was detected even when the FCFs were stored in a vehicle trunk for 1 h in summer and autumn and 2 h in spring and winter. Among three boxes (corrugated box, styrofoam box, and corrugated box coated with an aluminum film), the styrofoam box maintained the lowest temperature and showed the lowest growth rate of microorganisms on FCF after storage for 3 h in the vehicle trunk depending on whether ice was added. These results indicated that the possibility of food poisoning occurs when FCFs are exposed to the external environment. It is necessary to provide guidelines regarding storage temperature and allowable time for safe consumption of FCFs after purchase.

A Study on the Mechanisms by Which the Aqueous Extract of Inonotus obliquus Induces Apoptosis and Inhibits Proliferation in HT-29 Human Colon Cancer Cells (차가버섯 물추출물의 대장암세포 증식억제 및 Apoptosis 유도기전 연구)

  • Kim, Eun-Ji;Lee, Yong-Jin;Shim, Hyun-Kyung;YoonPark, Jung-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.5
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    • pp.516-523
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    • 2006
  • The mushroom Inonotus obliquue (IO) has been traditionally used for the treatment of gastrointestinal cancer in Russia, Poland, and most of Baltic countries. To explore the possibility that IO has chemoprevention effects, we examined whether or not the aqueous extract of IO inhibits HT-29 cell growth and investigated tile mechanism for this effect. Cells were incubated in the presence of increasing concentrations of the aqueous extract of IO. The extract substantially inhibited the viable HT-29 cell number in a dose-dependent manner and inhibited 5-bromo-2'-deoxyuridine incorporation into DNA of HT-29 cells. Annexin-V staining followed by flow cytometry revealed that the extract induced apoptosis of HT-29 cells in a dose-dependent manner. Western blot analysis of total cell lysates revealed that the extract induced cleavage of caspase-8, -9 and -3 and poly (ADP-ribose) polymerase, but did not affect the protein levels of Bax and Bcl-2. In addition, the extract dose-dependently increased the activity of caspase-8, -9 and -3. We have demonstrated that the aqueous extract of IO inhibits cell proliferation and induces apoptosis in HT-29 cells, which may be mediated by its ability to activate the caspase pathway.

Purification and Characterization of β-N-Acetylhexosaminidase from Rice Seeds

  • Jin, Yu-Lan;Jo, Yu-Young;Kim, Kil-Yong;Shim, Jae-Han;Kim, Yong-Woong;Park, Ro-Dong
    • BMB Reports
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    • v.35 no.3
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    • pp.313-319
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    • 2002
  • N-Acetyl-$\beta$-D-hexosaminidase ($\beta$-HexNAc'ase) (EC 3.2.1.52) was purified from rice seeds (Oryza sative L. var. Dongjin) using ammonium sulfate (80%) precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex chromatography, sequentially. The activities were separated into 7 fractions($F_1-F_7$) by CM-Sephadex chromatography. Among them, F6 was further purified to homogeneity with a 13.0% yield and 123.3 purification-fold. The molecular mass was estimated to be about 52 kDa on SDS-PAGE and 37.4 kDa on Sephacryl S-300 gel filtration. The enzyme catalyzed the hydrolysis of both p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GlcNAc) and p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GalNAc) as substrates, which are typical properties of $\beta$-HexNAc'ase. The ratio of the pNP-GlcNAc'ase activity to the pNP-GalNAc'ase activity was 4.0. However, it could not hydrolyze chitin, chitosan, pNP-$\beta$-glucopyranoside, or pNP-$\beta$-glucopyranoside. The enzyme showed $K_m$, $V_{max}$ and $K_{cat}$ for pNP-GlcNAc of 1.65 mM, $79.49\;mM\;min^{-1}$, and $4.79{\times}10^6\;min^{-1}$, respectively. The comparison of kinetic values for pNP-GlcNAc and pNP-GalNAc revealed that the two enzyme activities are associated with a single binding site. The purified enzyme exhibited optimum pH and temperature for pNP-GlcNAc of 5.0 and $50^{\circ}C$, respectively. The enzyme activity for pNP-GlcNAc was stable at pH 5.0-5.5 and $20-40^{\circ}C$. The enzyme activity was completely inhibited at a concentration of 0.1 mM $HgCl_$ and $AgNO_3$, suggesting that the intact thiol group is essential for activity. Chloramine T completely inhibited the activity, indicating the possible involvement of methionines in the mechanism of the enzyme.

Production and Characterization of Monoclonal Antibodies Specific to PAT Protein Expressed in Genetically Modified Herbicide-Resistance Maize (제초제 내성 유전자 변형 옥수수 중 PAT단백질에 특이한 단크론성 항체의 생산과 특성 확인)

  • Kim, Sol-A;Lee, Jeong-Eun;Shim, Won-Bo;Kang, Sung-Jo;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.193-199
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    • 2018
  • In this study, PAT protein of genetically modified maize was prepared from the recombinant E. coli strain BL21 (DE3), and mice were immunized with the recombinant PAT protein. After cell fusion and cloning, two hybridoma cells (PATmAb-7 and PATmAb-12) were chosen since the monoclonal antibodies (Mabs) produced by them were confirmed to be specific to PAT protein in the indirect enzyme-linked immunsorbent assay (ELISA) and western blot tests. There were no cross-reactions of either Mabs to other GM proteins or to the extracts of non-GM maize. The ELISA based on the PATmAb-7 can sensitively detect 0.3 ng/g PAT protein in corn. These results indicate that the developed Mabs can be used as bio-receptors in the development of immunosensors and biosensors for the rapid and simple detection of GM corn adulterated in foods.

A New Early Maturing, Lodging Resistance and High Yielding Vegetable Peanut "Seonan" (조숙 내도복 다수성 풋땅콩 품종 "선안")

  • Pae, Suk-Bok;Park, Chang-Hwan;Cheong, Young-Keun;Hwang, Chung-Dong;Lee, Myung-Hee;Shim, Kang-Bo;Jung, Chan-Sik;Kang, Churl-Whan;Park, Keum-Yong;Park, Chung-Berm;Choi, Gyu-Hwan;Lee, Jae-Chul;Kim, In-Jae;Kim, Sun-Lim;Kim, Je-Gyu
    • Korean Journal of Breeding Science
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    • v.40 no.2
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    • pp.178-183
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    • 2008
  • A new peanut variety "Seonan" (Arachis hypogaea ssp. hypogaea L.) was developed at the Yeongnam Agricultural Research Institute, NICS, in Milyang in 2007. It was developed from the cross between the erect short stem cultivar "SP9128" and the high-yielding cultivar "Daekwang". "Seanan" which is Virginia plant type has 18 branch number per plant with early maturing and long-ellipse shaped large kernel. Each pod has two grains with brown testa and 100 seed weight was 87 g in the regional yield trials (RYT). Especially this variety has resistance to lodging owing to short stem. The free sugar and tannin content of fresh peanut are 3.7% and 0.48%, respectively, similar to check variety. In the regional yield trials "Seonan" was outyielded than check variety by 14% with 8.59 MT/ha for fresh pod and by 7% with 3.75 MT/ha for grain.

Microbiological Hazard Analysis on Perilla Leaf Farms at the Harvesting Stage for the Application of the Good Agricultural Practices(GAP) (깻잎의 농산물우수관리제도(GAP) 적용을 위한 수확단계에서 미생물학적 위해요소 분석)

  • Kwon, Woo-Hyun;Lee, Won-Gyeong;Song, Jeong-Eon;Kim, Kyeong-Yeol;Shim, Won-Bo;Yoon, Yo-Han;Kim, Yun-Shik;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.295-300
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    • 2012
  • The purpose of this study was to analyze microbiological hazards for plants, cultivation environments and personal hygiene of perilla leaf farms at the harvesting stage. Samples were collected from three perilla leaf farms(A, B, C) located in Gyeongnam, Korea and tested for sanitary indications, fungi and pathogenic bacteria(Escherichia coli O157:H7, Listeria monocytogens, Salmonella spp., Staphylococcus aureus and Bacillus cereus). As a result, total bacteria and coliform in perilla leaf were detected at the levels of 4.4~5.2 and 3.4~4.3 log CFU/g, respectively, but E. coli was not detected in all samples. Among the pathogenic bacteria, B. cereus(perilla leaf: 2.0~2.4 log CFU/g, stem: 1.4~2.1 log CFU/g, water: 0.7 log CFU/ml, soil: 4.2~5.0 log CFU/g, hands: 3.0 log CFU/ hand, gloves: 2.1~2.4 log CFU/100 $cm^2$, glothes: 1.5~2.8 log CFU/100 $cm^2$) and S. aureus(3.4 log CFU/hand) were detected in all samples and worker's hand from farm A, respectively. However, other pathogenic bacteria were not detected. This study demonstrates that perilla leaf at the harvesting stage was significantly contaminated with microbial hazards.

Transforming growth factor-β promoted vascular endothelial growth factor release by human lung fibroblasts (인간 폐섬유아세포에서 TGF-β 자극에 의한 VEGF 분비)

  • Park, Sang-Uk;Shin, Joo-Hwa;Shim, Jae-Won;Kim, Deok-Soo;Jung, Hye-Lim;Park, Moon-Soo;Shim, Jung-Yeon
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.879-885
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    • 2008
  • Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.

The Correlation Analysis of Stress/Rest Ejection Fraction of $^{201}Tl$ Gated Myocardial Perfusion SPECT ($^{201}Tl$ 게이트 심근관류 스펙트에서의 휴식기와 부하기 좌심실 구혈률 상관관계 분석)

  • Kim, Dong-Seok;Yoo, Hee-Jae;Shim, Dong-Oh
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.3
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    • pp.3-9
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    • 2009
  • Purpose: It is well-known that stress-induced stunning and reversible perfusion defect have impact on ejection fraction (EF) when performing myocardial perfusion SPECT. Due to these reasons, gated SPECT is recommended at stress and rest studies. And there was many experiments to analyze between Stress and Rest EF by using $^{99m}Tc$-MIBI. The aim of this study is to analyze between stress EF and rest EF at myocardial perfusion SPECT by using $^{201}Tl$ and define possible predictors of EF variability. Materials and Methods: From 2008 June to 2009 February, we analyzed 144 patients undergoing $^{201}Tl$ gated myocardial perfusion SPECT in ASAN medical center. To analyze the data, we use QGS (Quantitative gated SPECT) software, and derived End-systolic volume (ESV), End-diastolic volume (EDV), EF from the result. In this study, we comparatively analyzed stress/rest EF correlation based on stress/rest EF, EDV, ESV and reversibility of myocardial perfusion defect by using paired t-test, Bland-Altman analysis. Results: Mached pairs of stress EF and rest EF demonstrated excellent correlation (r=0.92) with no statistically significant difference (p=0.11). Bland-Altman analysis demonstrated a mean ${\Delta}EF$ was 0.52% (95% confidential interval[CI], -1.17~0.12%). No statistically significant difference between a mean ${\Delta}EF$ and hypothetic mean of 0 (${\Delta}EF$=0) (p=0.10). In the correlation of ${\Delta}EF$ according to stress/rest EDV and ESV, except rest ESV of <28mL (p<0.05), there was no statistically significant difference. In the correlation of ${\Delta}EF$ according to reversibility of perfusion defect, patients with reversible perfusion defect has statistically significant difference of ${\Delta}EF$ (p<0.05). ${\Delta}EF$ of stress/rest EF showed no statistically significant difference except 55% of rest EF (p<0.05). Conclusion: Like studies with $^{99m}Tc$-MIBI, there was generally no statistically significant difference between stress and rest EF in this study results. However a stress EF of <55%, a rest ESV of <28mL and patients with reversible perfusion defect showed statistically significant difference in ${\Delta}EF$. If performing $^{201}T$ myocardial perfusion SPECT to patients with abnormal cardiac function or reversible perfusion defect, consider this study results and apply it. We expect this study results could be useful predictors of ${\Delta}EF$ variability.

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Association between cord blood 25-hydroxyvitamin D concentrations and respiratory tract infections in the first 6 months of age in a Korean population: a birth cohort study (COCOA)

  • Shin, Youn Ho;Yu, Jinho;Kim, Kyung Won;Ahn, Kangmo;Hong, Seo-Ah;Lee, Eun;Yang, Song-I;Jung, Young-Ho;Kim, Hyung Young;Seo, Ju-Hee;Kwon, Ji-Won;Kim, Byoung-Ju;Kim, Hyo-Bin;Shim, Jung Yeon;Kim, Woo Kyung;Song, Dae Jin;Lee, So-Yeon;Lee, Soo Young;Jang, Gwang Cheon;Suh, Dong In;Yang, Hyeon-Jong;Kim, Bong Sung;Hong, Soo-Jong
    • Clinical and Experimental Pediatrics
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    • v.56 no.10
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    • pp.439-445
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    • 2013
  • Purpose: Previous studies suggest that the concentration of 25-hydroxyvitamin D [25(OH)D] in cord blood may show an inverse association with respiratory tract infections (RTI) during childhood. The aim of the present study was to examine the influence of 25(OH)D concentrations in cord blood on infant RTI in a Korean birth cohort. Methods: The levels of 25(OH)D in cord blood obtained from 525 Korean newborns in the prospective COhort for Childhood Origin of Asthma and allergic diseases were examined. The primary outcome variable of interest was the prevalence of RTI at 6-month follow-up, as diagnosed by pediatricians and pediatric allergy and pulmonology specialists. RTI included acute nasopharyngitis, rhinosinusitis, otitis media, croup, tracheobronchitis, bronchiolitis, and pneumonia. Results: The median concentration of 25(OH)D in cord blood was 32.0 nmol/L (interquartile range, 21.4 to 53.2). One hundred and eighty neonates (34.3%) showed 25(OH)D concentrations less than 25.0 nmol/L, 292 (55.6%) showed 25(OH)D concentrations of 25.0-74.9 nmol/L, and 53 (10.1%) showed concentrations of ${\geq}75.0$ nmol/L. Adjusting for the season of birth, multivitamin intake during pregnancy, and exposure to passive smoking during pregnancy, 25(OH)D concentrations showed an inverse association with the risk of acquiring acute nasopharyngitis by 6 months of age (P for trend=0.0004). Conclusion: The results show that 89.9% of healthy newborns in Korea are born with vitamin D insufficiency or deficiency (55.6% and 34.3%, respectively). Cord blood vitamin D insufficiency or deficiency in healthy neonates is associated with an increased risk of acute nasopharyngitis by 6 months of age. More time spent outdoors and more intensified vitamin D supplementation for pregnant women may be needed to prevent the onset of acute nasopharyngitis in infants.