• Title/Summary/Keyword: RNA sequencing

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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.

Effect of ciglitazone on adipogenic transdifferentiation of bovine skeletal muscle satellite cells

  • Zhang, Junfang;Li, Qiang;Yan, Yan;Sun, Bin;Wang, Ying;Tang, Lin;Wang, Enze;Yu Jia;Nogoy, Kim Margarette Corpuz;Li, Xiangzi;Choi, Seong-Ho
    • Journal of Animal Science and Technology
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    • v.63 no.4
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    • pp.934-953
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    • 2021
  • Ciglitazone is a member of the thiazolidinedione family, and specifically binds to peroxisome proliferator-activated receptor-γ (PPARγ), thereby promoting adipocyte differentiation. We hypothesized that ciglitazone as a PPARγ ligand in the absence of an adipocyte differentiation cocktail would increase adiponectin and adipogenic gene expression in bovine satellite cells (BSC). Muscle-derived BSCs were isolated from six, 18-month-old Yanbian Yellow Cattle. The BSC were cultured for 96 h in differentiation medium containing 5 µM ciglitazone (CL), 10 µM ciglitazone (CM), or 20 µM ciglitazone (CH). Control (CON) BSC were cultured only in a differentiation medium (containing 2% horse serum). The presence of myogenin, desmin, and paired box 7 (Pax7) proteins was confirmed in the BSC by immunofluorescence staining. The CL, CM, and CH treatments produced higher concentrations of triacylglycerol and lipid droplet accumulation in myotubes than those of the CON treatment. Ciglitazone treatments significantly increased the relative expression of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, fatty acid synthase, stearoyl-CoA desaturase, and perilipin 2. Ciglitazone treatments increased gene expression of Pax3 and Pax7 and decreased expression of myogenic differentiation-1, myogenin, myogenic regulatory factor-5, and myogenin-4 (p < 0.01). Adiponectin concentration caused by ciglitazone treatments was significantly greater than CON (p < 0.01). RNA sequencing showed that 281 differentially expressed genes (DEGs) were found in the treatments of ciglitazone. DEGs gene ontology (GO) analysis showed that the top 10 GO enrichment significantly changed the biological processes such as protein trimerization, negative regulation of cell proliferation, adipocytes differentiation, and cellular response to external stimulus. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that DEGs were involved in the p53 signaling pathway, PPAR signaling pathway, biosynthesis of amino acids, tumor necrosis factor signaling pathway, non-alcoholic fatty liver disease, PI3K-Akt signaling pathway, and Wnt signaling pathway. These results indicate that ciglitazone acts as PPARγ agonist, effectively increases the adiponectin concentration and adipogenic gene expression, and stimulates the conversion of BSC to adipocyte-like cells in the absence of adipocyte differentiation cocktail.

In Vitro Tissue Culture Frequency and Transformation of Various Cultivars of Soybean (Glycine max (L.) Merr.) (다양한 콩 자원들의 기내 조직배양 효율 및 형질전환)

  • Seo, Mi-Suk;Cho, Chuloh;Jeong, Namhee;Sung, Soon-Kee;Choi, Man-Soo;Jin, Mina;Kim, Dool-Yi
    • Korean Journal of Plant Resources
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    • v.34 no.4
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    • pp.278-286
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    • 2021
  • Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean core-collection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.

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.

Expression and secretion of CXCL12 are enhanced in autosomal dominant polycystic kidney disease

  • Kim, Hyunho;Sung, Jinmo;Kim, Hyunsuk;Ryu, Hyunjin;Park, Hayne Cho;Oh, Yun Kyu;Lee, Hyun-Seob;Oh, Kook-Hwan;Ahn, Curie
    • BMB Reports
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    • v.52 no.7
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    • pp.463-468
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    • 2019
  • Autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic diseases (frequency of 1/1000-1/400), is characterized by numerous fluid-filled renal cysts (RCs). Inactivation of the PKD1 or PKD2 gene by germline and somatic mutations is necessary for cyst formation in ADPKD. To mechanistically understand cyst formation and growth, we isolated RCs from Korean patients with ADPKD and immortalized them with human telomerase reverse transcriptase (hTERT). Three hTERT-immortalized RC cell lines were characterized as proximal epithelial cells with germline and somatic PKD1 mutations. Thus, we first established hTERT-immortalized proximal cyst cells with somatic PKD1 mutations. Through transcriptome sequencing and Gene Ontology (GO) analysis, we found that upregulated genes were related to cell division and that downregulated genes were related to cell differentiation. We wondered whether the upregulated gene for the chemokine CXCL12 is related to the mTOR signaling pathway in cyst growth in ADPKD. CXCL12 mRNA expression and secretion were increased in RC cell lines. We then examined CXCL12 levels in RC fluids from patients with ADPKD and found increased CXCL12 levels. The CXCL12 receptor CXC chemokine receptor 4 (CXCR4) was upregulated, and the mTOR signaling pathway, which is downstream of the CXCL12/CXCR4 axis, was activated in ADPKD kidney tissue. To confirm activation of the mTOR signaling pathway by CXCL12 via CXCR4, we treated the RC cell lines with recombinant CXCL12 and the CXCR4 antagonist AMD3100; CXCL12 induced the mTOR signaling pathway, but the CXCR4 antagonist AMD3100 blocked the mTOR signaling pathway. Taken together, these results suggest that enhanced CXCL12 in RC fluids activates the mTOR signaling pathway via CXCR4 in ADPKD cyst growth.

Effect of commercially purified deoxynivalenol and zearalenone mycotoxins on microbial diversity of pig cecum contents

  • Reddy, Kondreddy Eswar;Kim, Minji;Kim, Ki Hyun;Ji, Sang Yun;Baek, Youlchang;Chun, Ju Lan;Jung, Hyun Jung;Choe, Changyong;Lee, Hyun Jeong;Kim, Minseok;Lee, Sung Dae
    • Animal Bioscience
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    • v.34 no.2
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    • pp.243-255
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    • 2021
  • Objective: Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that frequently contaminate maize and grain cereals, imposing risks to the health of both humans and animals and leading to economic losses. The gut microbiome has been shown to help combat the effects of such toxins, with certain microorganisms reported to contribute significantly to the detoxification process. Methods: We examined the cecum contents of three different dietary groups of pigs (control, as well as diets contaminated with 8 mg DON/kg feed or 0.8 mg ZEN/kg feed). Bacterial 16S rRNA gene amplicons were acquired from the cecum contents and evaluated by next-generation sequencing. Results: A total of 2,539,288 sequences were generated with ~500 nucleotide read lengths. Firmicutes, Bacteroidetes, and Proteobacteria were the dominant phyla, occupying more than 96% of all three groups. Lactobacillus, Bacteroides, Megasphaera, and Campylobacter showed potential as biomarkers for each group. Particularly, Lactobacillus and Bacteroides were more abundant in the DON and ZEN groups than in the control. Additionally, 52,414 operational taxonomic units were detected in the three groups; those of Bacteroides, Lactobacillus, Campylobacter, and Prevotella were most dominant and significantly varied between groups. Hence, contamination of feed by DON and ZEN affected the cecum microbiota, while Lactobacillus and Bacteroides were highly abundant and positively influenced the host physiology. Conclusion: Lactobacillus and Bacteroides play key roles in the process of detoxification and improving the immune response. We, therefore, believe that these results may be useful for determining whether disturbances in the intestinal microflora, such as the toxic effects of DON and ZEN, can be treated by modulating the intestinal bacterial flora.

Optimization of Medium to Improve Protease Production Using Response Surface Methodology by Bacillus amyloliquefaciens SRCM115785 (반응표면분석법을 이용한 Bacillus amyloliquefaciens SRCM115785의 protease 활성증가를 위한 배지 최적화)

  • Yang, Hee Gun;Ha, Gwangsu;Ryu, Myeong Seon;Park, Se Won;Jeong, Ho Jin;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.8
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    • pp.761-770
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    • 2021
  • In this study, the optimal medium composition for enhancing protease production was established by the Bacillus strain isolated from Makgeolli, a traditional fermented food, using the response surface methodology. B. amyloliquefaciens SRCM115785 was selected as the protease producer by productivity analysis and identified by 16S rRNA gene sequencing. Plackett-Burman design (PBD) was introduced to analyze the effect of each component on protease production among the 11 selected medium components. As a result, glucose, yeast extract, and beef extract were finally selected as factors for enhancing protease production. Central composite design (CCD) analysis was designed as a method to determine the optimal concentration of each component for protease production and the concentration of each medium composition for maximum protease production was predicted to glucose 6.75 g/l, yeast extract 12.42 g/l and beef extract 17.48 g/l. The suitability of the experimental model was proved using ANOVA analysis and as a result of quantitative analysis to prove this, the amount of increase was 230.47% compared to the LB medium used as a control. Through this study, the optimization of medium composition for enhancing protease production was established, and based on this, it is expected that it can be efficient use of protease as an industrial enzyme.

Biochemical properties and gluten degradation of Lactobacillus paracasei strain GLU70 isolated from salted seafood (젓갈에서 분리한 락토바실러스 파라카제이 GLU70 균주의 생화학적 특성 및 글루텐 분해능)

  • Park, Hyein;Yoon, Seul Gi;Jang, Junho;Byun, Ji Young;Yoon, Bok Kun
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.203-208
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    • 2022
  • Gluten is an insoluble protein present in cereals such as wheat. Gluten consumed through food is not digested and accumulates in the body; this has been linked to digestive discomfort, irritation, and various digestive disorders, including intestinal inflammation. In this study, the Lactobacillus paracasei strain GLU70, which exhibits a glutendegrading ability, was isolated from salted seafood. At a pH of 3.0, GLU70 showed a survival rate of approximately 84%, and at 0.3% oxgall, it showed a survival rate of approximately 53%. When the culture supernatant collected after 12 h of incubation was added to flour dough, approximately 50% gluten degradation was observed. Moreover, among several probiotic isolates exhibiting proteolytic activity selected to assess the gluten-degrading ability, GLU70 showed superior results regardless of the dough fermentation temperature. Although further research is required, GLU70 is expected to be of value in manufacturing gluten-reduced products and the food industry as an ingredient or additive.

Appropriate Soil Heat Treatment Promotes Growth and Disease Suppression of Panax notoginseng by Interfering with the Bacterial Community

  • Li, Ying-Bin;Zhang, Zhi-Ping;Yuan, Ye;Huang, Hui-Chuan;Mei, Xin-Yue;Du, Fen;Yang, Min;Liu, Yi-Xiang;Zhu, Shu-Sheng
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.294-301
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    • 2022
  • In our greenhouse experiment, soil heat treatment groups (50, 80, and 121℃) significantly promoted growth and disease suppression of Panax notoginseng in consecutively cultivated soil (CCS) samples (p < 0.01), and 80℃ worked better than 50℃ and 121℃ (p < 0.01). Furthermore, we found that heat treatment at 80℃ changes the microbial diversity in CCS, and the inhibition ratios of culturable microorganisms, such as fungi and actinomycetes, were nearly 100%. However, the heat-tolerant bacterial community was preserved. The 16S rRNA gene and internal transcribed spacer (ITS) sequencing analyses indicated that the soil heat treatment had a greater effect on the Chao1 index and Shannon's diversity index of bacteria than fungi, and the relative abundances of Firmicutes and Proteobacteria were significantly higher than without heating (80 and 121℃, p < 0.05). Soil probiotic bacteria, such as Bacillus (67%), Sporosarcina (9%), Paenibacillus (6%), Paenisporosarcina (6%), and Cohnella (4%), remained in the soil after the 80℃ and 121℃ heat treatments. Although steam increased the relative abundances of most of the heat-tolerant microbes before sowing, richness and diversity gradually recovered to the level of CCS, regardless of fungi or bacteria, after replanting. Thus, we added heat-tolerant microbes (such as Bacillus) after steaming, which reduced the relative abundance of pathogens, recruited antagonistic bacteria, and provided a long-term protective effect compared to the steaming and Bacillus alone (p < 0.05). Taken together, the current study provides novel insight into sustainable agriculture in a consecutively cultivated system.

Simotang Alleviates the Gastrointestinal Side Effects of Chemotherapy by Altering Gut Microbiota

  • Deng, Lijing;Zhou, Xingyi;Lan, Zhifang;Tang, Kairui;Zhu, Xiaoxu;Mo, Xiaowei;Zhao, Zongyao;Zhao, Zhiqiang;Wu, Mansi
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.405-418
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    • 2022
  • Simotang oral liquid (SMT) is a traditional Chinese medicine (TCM) consisting of four natural plants and is used to alleviate gastrointestinal side effects after chemotherapy and functional dyspepsia (FD). However, the mechanism by which SMT helps cure these gastrointestinal diseases is still unknown. Here, we discovered that SMT could alleviate gastrointestinal side effects after chemotherapy by altering gut microbiota. C57BL/6J mice were treated with cisplatin (DDP) and SMT, and biological samples were collected. Pathological changes in the small intestine were observed, and the intestinal injury score was assessed. The expression levels of the inflammatory factors IL-1β and IL-6 and the adhesive factors Occludin and ZO-1 in mouse blood or small intestine tissue were also detected. Moreover, the gut microbiota was analyzed by high-throughput sequencing of 16S rRNA amplicons. SMT was found to effectively reduce gastrointestinal mucositis after DDP injection, which lowered inflammation and tightened the intestinal epithelial cells. Gut microbiota analysis showed that the abundance of the anti-inflammatory microbiota was downregulated and that the inflammatory microbiota was upregulated in DDP-treated mice. SMT upregulated anti-inflammatory and anticancer microbiota abundance, while the inflammatory microbiota was downregulated. An antibiotic cocktail (ABX) was also used to delete mice gut microbiota to test the importance of gut microbiota, and we found that SMT could not alleviate gastrointestinal mucositis after DDP injection, showing that gut microbiota might be an important mediator of SMT treatment. Our study provides evidence that SMT might moderate gastrointestinal mucositis after chemotherapy by altering gut microbiota.