• Title/Summary/Keyword: Profiling Methods

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Differential MicroRNA Expression Between Gastric Cancer Tissue and Non-cancerous Gastric Mucosa According to Helicobacter pylori Status

  • Lee, Jung Won;Kim, Nayoung;Park, Ji Hyun;Kim, Hee Jin;Chang, Hyun;Kim, Jung Min;Kim, Jin-Wook;Lee, Dong Ho
    • Journal of Cancer Prevention
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    • v.22 no.1
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    • pp.33-39
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    • 2017
  • Background: MicroRNAs (miRNAs) are key post-translational mechanisms which can regulate gene expression in gastric carcinogenesis. To identify miRNAs responsible for gastric carcinogenesis, we compared expression levels of miRNAs between gastric cancer tissue and non-cancerous gastric mucosa according to Helicobacter pylori status. Methods: Total RNA was extracted from the cancerous regions of formalin-fixed, paraffin-embedded tissues of H. pylori-positive (n = 8) or H. pylori-negative (n = 8) patients with an intestinal type of gastric cancer. RNA expression was analyzed using a 3,523 miRNA profiling microarray based on the Sanger miRBase. Validation analysis was performed using TaqMan miRNA assays for biopsy samples from 107 patients consisted of control and gastric cancer with or without H. pylori. And then, expression levels of miRNAs were compared according to subgroups. Results: A total of 156 miRNAs in the aberrant miRNA profiles across the miRNA microarray showed differential expression (at least a 2-fold change, P < 0.05) in cancer tissue, compared to noncancerous mucosa in both of H. pylori-negative and -positive samples. After 10 promising miRNAs were selected, validations by TaqMan miRNA assays confirmed that two miRNAs (hsa-miR-135b-5p and hsa-miR-196a-5p) were significantly increased and one miRNA (hsa-miR-145-5p) decreased in cancer tissue compared to non-cancerous gastric mucosa at H. pylori-negative group. For H. pylori-positive group, three miRNAs (hsa-miR-18a-5p, hsa-miR-135b-5p, and hsa-miR-196a-5p) were increased in cancer tissue. hsa-miR-135b-5p and hsa-miR-196a-5p were increased in gastric cancer in both of H. pylori-negative and -positive. Conclusions: miRNA expression of the gastric cancer implies that different but partially common gastric cancer carcinogenic mechanisms might exist according to H. pylori status.

Effects on microbial diversity of fermentation temperature (10℃ and 20℃), long-term storage at 5℃, and subsequent warming of corn silage

  • Zhou, Yiqin;Drouin, Pascal;Lafreniere, Carole
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1528-1539
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    • 2019
  • Objective: To evaluate the effects on microbial diversity and biochemical parameters of gradually increasing temperatures, from $5^{\circ}C$ to $25^{\circ}C$ on corn silage which was previously fermented at ambient or low temperature. Methods: Whole-plant corn silage was fermented in vacuum bag mini-silos at either $10^{\circ}C$ or $20^{\circ}C$ for two months and stored at $5^{\circ}C$ for two months. The mini-silos were then subjected to additional incubation from $5^{\circ}C$ to $25^{\circ}C$ in $5^{\circ}C$ increments. Bacterial and fungal diversity was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiling and biochemical analysis from mini-silos collected at each temperature. Results: A temperature of $10^{\circ}C$ during fermentation restricted silage fermentation compared to fermentation temperature of $20^{\circ}C$. As storage temperature increased from $5^{\circ}C$ to $25^{\circ}C$, little changes occurred in silages fermented at $20^{\circ}C$, in terms of most biochemical parameters as well as bacterial and fungal populations. However, a high number of enterobacteria and yeasts (4 to $5\;log_{10}$ colony forming unit/g fresh materials) were detected at $15^{\circ}C$ and above. PCR-DGGE profile showed that Candida humilis predominated the fungi flora. For silage fermented at $10^{\circ}C$, no significant changes were observed in most silage characteristics when temperature was increased from $5^{\circ}C$ to $20^{\circ}C$. However, above $20^{\circ}C$, silage fermentation resumed as observed from the significantly increased number of lactic acid bacteria colonies, acetic acid content, and the rapid decline in pH and water-soluble carbohydrates concentration. DGGE results showed that Lactobacillus buchneri started to dominate the bacterial flora as temperature increased from $20^{\circ}C$ to $25^{\circ}C$. Conclusion: Temperature during fermentation as well as temperature during storage modulates microorganism population development and fermentation patterns. Silage fermented at $20^{\circ}C$ indicated that these silages should have lower aerobic stability at opening because of better survival of yeasts and enterobacteria.

Xiang Study: an association of breastmilk composition with maternal body mass index and infant growth during the first 3 month of life

  • Peng, Xuyi;Li, Jie;Yan, Shuyuan;Chen, Juchun;Lane, Jonathan;Malard, Patrice;Liu, Feitong
    • Nutrition Research and Practice
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    • v.15 no.3
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    • pp.367-381
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    • 2021
  • BACKGROUND/OBJECTIVES: This study aimed to establish a mother and child cohort in the Chinese population, and investigate human breastmilk (HBM) composition and its relationship with maternal body mass index (BMI) and infant growth during the first 3 mon of life. SUBJECTS/METHODS: A total of 101 Chinese mother and infant pairs were included in this prospective cohort. Alterations in the milk macronutrients of Chinese mothers at 1 mon (T1), 2 mon (T2), and 3 mon (T3) lactation were analyzed. HBM fatty acid (FA) profiles were measured by gas chromatography (GC), and HBM proteomic profiling was achieved by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). RESULTS: During the first 3 mon of lactation (P < 0.05), significant decreases were determined in the levels of total energy, fat, protein, and osteopontin (OPN), as well as ratios of long-chain saturated FA (including C16:0, C22:0 and C24:0), monounsaturated FA (including C16:1), and n-6 poly unsaturated FA (PUFA) (including C20:3n-6 and C20:4n-6, and n-6/n-3). Conversely, butyrate, C6:0 and n-3 PUFA C18:3n-3 (α-linolenic acid, ALA) were significantly increased during the first 3 mon (P < 0.05). HBM proteomic analyses distinguished compositional protein differences over time (P = 0.001). Personalized motherinfant analyses demonstrated that HBM from high BMI mothers presented increased total energy, fat, protein and OPN, and increased content of n-6 PUFA (including C18:3n-6, C20:3n-6 and n-6/n-3 ratio) as compared with low BMI mothers (P < 0.05). Furthermore, BMI of the mothers positively correlated with the head circumference (HC) of infants as well as the specific n-6 PUFA C20:3n-6 over the 3 time points examined. Infant HC was negatively associated with C18:0. CONCLUSION: This study provides additional evidence to the Chinese HBM database, and further knowledge of FA function. It also helps to establish future maternal strategies that support the healthy growth and development of Chinese infants.

Quantifying Chloride Ingress in Cracked Concrete Using Image Processing (이미지 분석을 이용한 균열 콘크리트 내 염화물 침투 정량화 평가)

  • Kim, Kun-Soo;Park, Ki-Tae;Kim, Jaehwan
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.26 no.4
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    • pp.57-64
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    • 2022
  • Chloride, which is one of the main deterioration factors in reinforced concrete structures, can degrade the performance of the structure due to chloride-induced corrosion of steel. Chloride content at steel depth or the rate of chloride penetration is necessary to determine deterioration of reinforced concrete or to calculate initiation time of steel corrosion caused by chloride attack. Chlorides in concrete are generally identified with typical two methods including chloride profiling using potentiometric titration method and discoloration method using AgNO3 solution. The former is advantageous to estimate chloride penetration rate (diffusion coefficient in general) with measured chloride contents directly, but it is laborious. In the case of latter, while the result is obtained easily with the range of discoloration, the error may occur depending on workmanship when the depth of chloride ingress is measured. This study shows that chloride penetrated depth is evaluated with the results obtained from discoloration method through image analysis, thereby the error is minimized by workmanship. In addition, the effect of micro-crack in concrete is studied on chloride penetration. In conclusion, the depth of chloride penetration was quantified with image analysis and as it was confirmed that chlorides can rapidly penetrate through micro-cracks, caution is especially required for cracks in concrete structure.

Anti-inflammatory Effect of Houttuyniae Herba Water Extract on LPS-induced RAW 264.7 Mouse Macrophages (마우스 대식세포를 이용한 어성초(魚腥草) 물추출물의 항염효능연구)

  • Hwang, In Seung;Kim, Young Jin;Park, Yun Soo;Kim, Hyun Ju;Kim, Do Hoon;Park, Wan Su
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.83-89
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    • 2014
  • Objectives : The purpose of this study was to investigate effects of Houttuyniae Herba water extract (HC) on calcium release and production of various inflammatory mediators such as nitric oxide (NO), interferon-inducible protein (IP)-10, platelet derived growth factor (PDGF)-BB, keratinocyte-derived chemokine (KC), vascular endothelial growth factor (VEGF), interleukin (IL)-4, and IL-5 in lipopolysaccharide (LPS)-induced RAW 264.7 mouse macrophages. Methods : NO production was measured by Griess reagent assay. Intracellular calcium level was measured with Fluo-4 assay. Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. Results : HC significantly decreased NO production for 24 hrs incubation at the concentrations of 10, 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC significantly decreased production of IP-10, KC, VEGF, and PDGF-BB for 24 hrs incubation at the concentrations of 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC also significantly decreased intracellular calcium release for 24 hrs incubation at the concentrations of 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). But HC did not show any significant effect on production of IL-4 and IL-5 in LPS-induced RAW 264.7. Conclusions : The results suggested that HC has anti-inflammatory property related with its inhibition on the production of NO, IP-10, KC, VEGF, and PDGF-BB in LPS-induced macrophages via calcium pathway.

Comparative transcriptome and metabolome analyses of four Panax species explore the dynamics of metabolite biosynthesis

  • Hyunjin, Koo;Yun Sun, Lee;Van Binh, Nguyen;Vo Ngoc Linh, Giang;Hyun Jo, Koo;Hyun-Seung, Park;Padmanaban, Mohanan;Young Hun, Song;Byeol, Ryu;Kyo Bin, Kang;Sang Hyun, Sung;Tae-Jin, Yang
    • Journal of Ginseng Research
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    • v.47 no.1
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    • pp.44-53
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    • 2023
  • Background: The genus Panax in the Araliaceae family has been used as traditional medicinal plants worldwide and is known to biosynthesize ginsenosides and phytosterols. However, genetic variation between Panax species has influenced their biosynthetic pathways is not fully understood. Methods: Simultaneous analysis of transcriptomes and metabolomes obtained from adventitious roots of two tetraploid species (Panax ginseng and P. quinquefolius) and two diploid species (P. notoginseng and P. vietnamensis) revealed the diversity of their metabolites and related gene expression profiles. Results: The transcriptome analysis showed that 2,3-OXIDOSQUALENE CYCLASEs (OSCs) involved in phytosterol biosynthesis are upregulated in the diploid species, while the expression of OSCs contributing to ginsenoside biosynthesis is higher in the tetraploid species. In agreement with these results, the contents of dammarenediol-type ginsenosides were higher in the tetraploid species relative to the diploid species. Conclusion: These results suggest that a whole-genome duplication event has influenced the triterpene biosynthesis pathway in tetraploid Panax species during their evolution or ecological adaptation. This study provides a basis for further efforts to explore the genetic variation of the Panax genus.

Transcriptome profiling identifies immune response genes against porcine reproductive and respiratory syndrome virus and Haemophilus parasuis co-infection in the lungs of piglets

  • Zhang, Jing;Wang, Jing;Zhang, Xiong;Zhao, Chunping;Zhou, Sixuan;Du, Chunlin;Tan, Ya;Zhang, Yu;Shi, Kaizhi
    • Journal of Veterinary Science
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    • v.23 no.1
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    • pp.2.1-2.18
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    • 2022
  • Background: Co-infections of the porcine reproductive and respiratory syndrome virus (PRRSV) and the Haemophilus parasuis (HPS) are severe in Chinese pigs, but the immune response genes against co-infected with 2 pathogens in the lungs have not been reported. Objectives: To understand the effect of PRRSV and/or HPS infection on the genes expression associated with lung immune function. Methods: The expression of the immune-related genes was analyzed using RNA-sequencing and bioinformatics. Differentially expressed genes (DEGs) were detected and identified by quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and western blotting assays. Results: All experimental pigs showed clinical symptoms and lung lesions. RNA-seq analysis showed that 922 DEGs in co-challenged pigs were more than in the HPS group (709 DEGs) and the PRRSV group (676 DEGs). Eleven DEGs validated by qRT-PCR were consistent with the RNA sequencing results. Eleven common Kyoto Encyclopedia of Genes and Genomes pathways related to infection and immune were found in single-infected and co-challenged pigs, including autophagy, cytokine-cytokine receptor interaction, and antigen processing and presentation, involving different DEGs. A model of immune response to infection with PRRSV and HPS was predicted among the DEGs in the co-challenged pigs. Dual oxidase 1 (DUOX1) and interleukin-21 (IL21) were detected by IHC and western blot and showed significant differences between the co-challenged pigs and the controls. Conclusions: These findings elucidated the transcriptome changes in the lungs after PRRSV and/or HPS infections, providing ideas for further study to inhibit ROS production and promote pulmonary fibrosis caused by co-challenging with PRRSV and HPS.

Study on the Applicability of CPT Based Soil Classification Chart (콘관입시험결과를 이용한 흙분류차트의 적용성에 관한 연구)

  • Kim, Chan-Hong;Im, Jong-Chul;Kim, Young-Sang
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.28 no.5C
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    • pp.293-301
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    • 2008
  • Soil profiling is one of the most important work among geotehnical engineering practice. Generally, soil profile is estimated from the observation of soil samples during subsurface exploration but such estimation also includes some experiencing aspects such as flushed water from the borehole, slime colour, boring speed and so on. In addition, since the capacity of hydraulic drill rig is significantly increased, thin layers might be easily missed. So, continuous soil profile is almost impossible over all depth to be bored from conventional subsurface exploration. While CPT or CPTu can serve continuous soil profile information over all depth generally in 5cm interval. Many charts or methods for soil profile from CPT result have been proposed during last several decades over the world. However they have not been verified in local ground condition in Korea. In this research, CPT results and soil classification results based on USCS were compiled from 17 sites over the Korea. Soil classification results by using 7 CPT soil classification charts were compared with those of USCS for the compiled database. Most proper CPT soil classification chart for Korean soil characteristics was evaluated and effective parameters for the soil classification from CPT were discussed. Finally interrelationship between CPT soil classification chart and USCS soil classification was evaluated.

RNA helicase DEAD-box-5 is involved in R-loop dynamics of preimplantation embryos

  • Hyeonji Lee;Dong Wook Han;Seonho Yoo;Ohbeom Kwon;Hyeonwoo La;Chanhyeok Park;Heeji Lee;Kiye Kang;Sang Jun Uhm;Hyuk Song;Jeong Tae Do;Youngsok Choi;Kwonho Hong
    • Animal Bioscience
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    • v.37 no.6
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    • pp.1021-1030
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    • 2024
  • Objective: R-loops are DNA:RNA triplex hybrids, and their metabolism is tightly regulated by transcriptional regulation, DNA damage response, and chromatin structure dynamics. R-loop homeostasis is dynamically regulated and closely associated with gene transcription in mouse zygotes. However, the factors responsible for regulating these dynamic changes in the R-loops of fertilized mouse eggs have not yet been investigated. This study examined the functions of candidate factors that interact with R-loops during zygotic gene activation. Methods: In this study, we used publicly available next-generation sequencing datasets, including low-input ribosome profiling analysis and polymerase II chromatin immunoprecipitation-sequencing (ChIP-seq), to identify potential regulators of R-loop dynamics in zygotes. These datasets were downloaded, reanalyzed, and compared with mass spectrometry data to identify candidate factors involved in regulating R-loop dynamics. To validate the functions of these candidate factors, we treated mouse zygotes with chemical inhibitors using in vitro fertilization. Immunofluorescence with an anti-R-loop antibody was then performed to quantify changes in R-loop metabolism. Results: We identified DEAD-box-5 (DDX5) and histone deacetylase-2 (HDAC2) as candidates that potentially regulate R-loop metabolism in oocytes, zygotes and two-cell embryos based on change of their gene translation. Our analysis revealed that the DDX5 inhibition of activity led to decreased R-loop accumulation in pronuclei, indicating its involvement in regulating R-loop dynamics. However, the inhibition of histone deacetylase-2 activity did not significantly affect R-loop levels in pronuclei. Conclusion: These findings suggest that dynamic changes in R-loops during mouse zygote development are likely regulated by RNA helicases, particularly DDX5, in conjunction with transcriptional processes. Our study provides compelling evidence for the involvement of these factors in regulating R-loop dynamics during early embryonic development.

Correlation between MR Image-Based Radiomics Features and Risk Scores Associated with Gene Expression Profiles in Breast Cancer (유방암에서 자기공명영상 근거 영상표현형과 유전자 발현 프로파일 근거 위험도의 관계)

  • Ga Ram Kim;You Jin Ku;Jun Ho Kim;Eun-Kyung Kim
    • Journal of the Korean Society of Radiology
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    • v.81 no.3
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    • pp.632-643
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    • 2020
  • Purpose To investigate the correlation between magnetic resonance (MR) image-based radiomics features and the genomic features of breast cancer by focusing on biomolecular intrinsic subtypes and gene expression profiles based on risk scores. Materials and Methods We used the publicly available datasets from the Cancer Genome Atlas and the Cancer Imaging Archive to extract the radiomics features of 122 breast cancers on MR images. Furthermore, PAM50 intrinsic subtypes were classified and their risk scores were determined from gene expression profiles. The relationship between radiomics features and biomolecular characteristics was analyzed. A penalized generalized regression analysis was performed to build prediction models. Results The PAM50 subtype demonstrated a statistically significant association with the maximum 2D diameter (p = 0.0189), degree of correlation (p = 0.0386), and inverse difference moment normalized (p = 0.0337). Among risk score systems, GGI and GENE70 shared 8 correlated radiomic features (p = 0.0008-0.0492) that were statistically significant. Although the maximum 2D diameter was most significantly correlated to both score systems (p = 0.0139, and p = 0.0008), the overall degree of correlation of the prediction models was weak with the highest correlation coefficient of GENE70 being 0.2171. Conclusion Maximum 2D diameter, degree of correlation, and inverse difference moment normalized demonstrated significant relationships with the PAM50 intrinsic subtypes along with gene expression profile-based risk scores such as GENE70, despite weak correlations.