• 생명과학회지
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• 제22권5호
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• pp.621-626
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• 2012

초파리는 발생과 질병연구를 위한 모델 곤충으로 널리 이용되어 왔다. 본 연구에서도 초파리를 모델곤충으로 한 장질환 연구의 일환으로 다양한 병원균 감염 및 장질환 유발시 어떻게 장줄기세포가 작용하는지를 이해하기 위해 초파리 장세포의 일차배양 방법을 확립하였다. 초파리 성충으로부터 장을 해부하고 다양한 효소를 처리하여 장세포를 분리한 후 배양하였다. 배양세포의 생존여부는 현미경 검경 및 MTS assay에 의해 확인한 결과 배양 후 9일째 최대 증식되었고 14일까지 생존함을 확인할 수 있었다. 또한 장줄기세포 및 장내분비세포의 존재도 immunostaining에 의해 확인하였다. 따라서 본 연구에서 구축된 초파리 일차배양 장세포는 다양한 유전자에 의한 장줄기 세포 조절연구뿐만 아니라 장에서 발생하는 다양한 질병을 연구하는 도구로 매우 유용할 것으로 추측된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권11호
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• pp.1659-1664
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• 2008

To study the working mechanisms for non-starch polysaccharidases to improve the growth performance of broiler chickens, a 21-day feeding trial was conducted. Two dietary treatments were included: 1) wheat diet (the control); 2) wheat+xylanase diet (xylanase, Allzyme PT, Alltech, Kentucky, USA). There were 8 replicates with 8 birds each for each treatment and the experimental diets were given to birds from hatch. Feed intake and body weight were measured on days 7 and 21. At the same ages, samples were taken for the determination of selected groups of luminal and mucosa-associated bacteria, mucosal morphology, brush-border membrane (BBM) bound enzyme activity and ileal nutrient digestibility. The xylanase supplement increased (p<0.05) body weight gain (BWG) and improved feed conversion ratio (FCR) at the end of the experiment but protein and starch digestibilities were not affected (p>0.05) by xylanase. Up to day 7, xylanase increased the counts of C. perfringens in the ileum and total anaerobic bacteria (TAB) in the caeca (p<0.05, p=0.07, respectively). By day 21, the counts of ileal lactobacilli (p<0.05) and TAB (p=0.07) were lower in birds given the xylanase-supplemented diet than in those on the control diet. No significant differences were observed in the counts of mucosa-associated lactobacilli and coliforms between xylanase treatment and the control at both ages. Villus height at the jejunum was not affected (p>0.05) by the supplement but crypt depth at the same site was reduced at day 7. Also, xylanase tended to increase the concentration of BBM protein (p = 0.09) and the specific activity of sucrase (p = 0.07) at day 21.

• Archives of Pharmacal Research
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• 제24권3호
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• pp.229-233
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• 2001

This Paper deals with the development of a technology for making a hydrophilic gel of Polyethylene oxide reception in which radiating ability is employed to cause cross-linking of Polymers in a water solution. The gel of polyethylene oxide was shown to be nontoxic contain 5-50% of polymer and be useful in composite medicinal forms along with biologically active substances including Bac. subtilis proteases. Proteases immobilized in the gel possess high thermal stability and proteolytic activity and are readily applied in medicine. The effect of immobilized proteolytic and glucolytic enzymes of Bac. subtillis (Immozimase) on the warm ischemia-reperfusion (I/R) which can cause hepatic and jejunum injury was also studied. These enzymes were immobilized on water-soluble polymer polyethylene glycol by means of an electron beam. The number of degraanulated mast cells as well as serum ALT after I/R in the group with Immozimase was decreased to almost half as compared with the control group. Pretreatment with Immozimase resulted in significant reduction of hepatic and gut neutrophil accumulation as compared with control animals. It was concluded that Immozimase has a protective effect for hepatic and gut ischemia/reperfusion, and this effect seems to be associated with prevention of leukocyte accumulation .

• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1144-1172
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• 2022

Salmonella enterica serovar Typhimurium isolate HJL777 is a virulent bacterial strain in pigs. The high rate of salmonella infection are at high risk of non-typhoidal salmonella gastroenteritis development. Salmonellosis is most common in young pigs. We investigated changes in gut microbiota and biological function in piglets infected with salmonella via analysis of rectal fecal metagenome and intestinal transcriptome using 16S rRNA and RNA sequencing. We identified a decrease in Bacteroides and increase in harmful bacteria such as Spirochaetes and Proteobacteria by microbial community analysis. We predicted that reduction of Bacteroides by salmonella infection causes proliferation of salmonella and harmful bacteria that can cause an intestinal inflammatory response. Functional profiling of microbial communities in piglets with salmonella infection showed increasing lipid metabolism associated with proliferation of harmful bacteria and inflammatory responses. Transcriptome analysis identified 31 differentially expressed genes. Using gene ontology and Innate Immune Database analysis, we identified that BGN, DCN, ZFPM2 and BPI genes were involved in extracellular and immune mechanisms, specifically salmonella adhesion to host cells and inflammatory responses during infection. We confirmed alterations in gut microbiota and biological function during salmonella infection in piglets. Our findings will help prevent disease and improve productivity in the swine industry.

• Journal of Microbiology and Biotechnology
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• 제32권11호
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• pp.1416-1426
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• 2022

The need to discover new types of antimicrobial agents has grown since the emergence of antibiotic-resistant pathogens that threaten human health. The world's oceans, comprising complex niches of biodiversity, are a promising environment from which to extract new antibiotics-like compounds. In this study, we newly isolated Pseudomonas sp. NIBR-H-19 from the gut of the sea roach Ligia exotica and present both phenotypes and genomic information consisting of 6,184,379 bp in a single chromosome possessing a total of 5,644 protein-coding genes. Genomic analysis of the isolated species revealed that numerous genes involved in antimicrobial secondary metabolites are predicted throughout the whole genome. Moreover, our analysis showed that among twenty-five pathogenic bacteria, the growth of three pathogens, including Staphylococcus aureus, Streptococcus hominis and Rhodococcus equi, was significantly inhibited by the culture of Pseudomonas sp. NIBR-H-19. The characterization of marine microorganisms with biochemical assays and genomics tools will help uncover the biosynthesis and action mechanism of antimicrobial metabolites for development as antagonistic probiotics against fish pathogens in an aquatic culture system.

• Journal of Microbiology and Biotechnology
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• 제33권8호
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• pp.1057-1065
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• 2023

Inflammatory bowel disease (IBD), a chronic inflammatory disease, results from dysregulation of the immune responses. Some lactic acid bacteria (LAB), including Lactobacillus, alleviate IBD through immunomodulation. In this study, the anti-colitis effect of LAB isolated from human breast milk was investigated in a mouse model induced acute colitis with 2,4,6-trinitrobenzene sulfonic acid (TNBS). TNBS remarkably increased weight loss, colon shortening, and colonic mucosal proliferation, as well as the expression levels of inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-1β. Oral administration of LAB isolated from human breast milk resulted in a reduction in TNBS-induced colon shortening, as well as induced cyclooxygenase (COX)-2, nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB). In addition, LAB suppressed inflammatory cytokines such as TNF-α, IL-6, and IL-1β, and thus showed an effect of suppressing the level of inflammation induced by TNBS. Furthermore, LAB alleviated gut microbiota dysbiosis, and inhibited intestinal permeability by increasing the expression of intestinal tight junction protein including ZO-1. Collectively, these results suggest that LAB isolated from human breast milk can be used as a functional food for colitis treatment by regulating NF-κB signaling, gut microbiota and increasing expression of intestinal tight junction protein.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1149-1161
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• 2023

Changes in the gut microbiome cause recolonization by pathogens and inflammatory responses, leading to the development of intestinal disorders. Probiotics administration has been proposed for many years to reverse the intestinal dysbiosis and to enhance intestinal health. This study aimed to evaluate the inhibitory effects of two newly designed probiotic mixtures, Consti-Biome and Sensi-Biome, on two enteric pathogens Staphylococcus aureus and Escherichia coli that may cause intestinal disorders. Additionally, the study was designed to evaluate whether Consti-Biome and Sensi-Biome could modulate the immune response, produce short-chain fatty acids (SCFAs), and reduce gas production. Consti-Biome and Sensi-Biome showed superior adhesion ratios to HT-29 cells and competitively suppressed pathogen adhesion. Moreover, the probiotic mixtures decreased the levels of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-6 and IL-1β. Cell-free supernatants (CFSs) were used to investigate the inhibitory effects of metabolites on growth and biofilms of pathogens. Consti-Biome and Sensi-Biome CFSs exhibited antimicrobial and anti-biofilm activity, where microscopic analysis confirmed an increase in the number of dead cells and the structural disruption of pathogens. Gas chromatographic analysis of the CFSs revealed their ability to produce SCFAs, including acetic, propionic, and butyric acid. SCFA secretion by probiotics may demonstrate their potential activities against pathogens and gut inflammation. In terms of intestinal symptoms regarding abdominal bloating and discomfort, Consti-Biome and Sensi-Biome also inhibited gas production. Thus, these two probiotic mixtures have great potential to be developed as dietary supplements to alleviate the intestinal disorders.

• Journal of Microbiology and Biotechnology
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• 제33권11호
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• pp.1506-1512
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• 2023

Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r² = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

• Fisheries and Aquatic Sciences
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• 제10권3호
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• pp.119-126
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• 2007

The analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and the development of resources for functional genomics. To analyze the transcriptome of the abalone Haliotis discus hannai, we conducted EST analysis using seven cDNA libraries made from gill, gut, hepatopancreas, skin, muscle, testis, and ovary. Redundant ESTs were assembled into overlapping contiguous sequences using the assembly program ICAtools. We found that the total 1,393 ESTs formed 135 clusters and 951 singletons, indicating that the overall redundancy of the library was 22%. Of the 1,393 clones, BLAST identified 1,278 clones (91.7%) as known genes; 115 clones (8.3%) did not match any previously described gene. Based on the major functions of their encoded proteins, the identified clones were classified into 16 broad categories. Sequence analysis revealed the presence of micro satellite-containing genes that may be valuable for further gene mapping studies. This study contributes to the identification of numerous EST clones that can be applied to further clarifying the genetics and developmental biology of abalone.