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Lactic Acid Bacteria Improves Peyer's Patch Cell-Mediated Immunoglobulin A and Tight-Junction Expression in a Destructed Gut Microbial Environment

  • Kim, Sung Hwan;Jeung, Woonhee;Choi, Il-Dong;Jeong, Ji-Woong;Lee, Dong Eun;Huh, Chul-Sung;Kim, Geun-Bae;Hong, Seong Soo;Shim, Jae-Jung;Lee, Jung Lyoul;Sim, Jae-Hun;Ahn, Young-Tae
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1035-1045
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    • 2016
  • To evaluate the effects of lactic acid bacteria (LAB) on Peyer's patch cells, mice were treated with a high dose of kanamycin to disturb the gut microbial environment. The overarching goal was to explore the potential of LAB for use as a dietary probiotic that buffers the negative consequences of antibiotic treatment. In vitro, LAB stimulated the production of immunoglobulin A (IgA) from isolated Peyer's patch cells. Inflammation-related genes (TNF-α, IL-1β, and IL-8) were up-regulated in Caco-2 cells stimulated with lipopolysaccharide (LPS), while tight-junction-related genes (ZO-1 and occludin) were down-regulated; the effects of LPS on inflammatory gene and tight-junction gene expression were reversed by treatment with LAB. Mice treated with a high dose of kanamycin showed increased serum IgE levels and decreases in serum IgA and fecal IgA levels; the number of Peyer's patch cells decreased with kanamycin treatment. However, subsequent LAB treatment was effective in reducing the serum IgE level and recovering the serum IgA and fecal IgA levels, as well as the number of Peyer's patch cells. In addition, ZO-1 and occludin mRNA levels were up-regulated in the ileum tissues of mice receiving LAB treatment. Lactic acid bacteria can enhance the intestinal immune system by improving the integrity of the intestinal barrier and increasing the production of IgA in Peyer's patches. Lactic acid bacteria should be considered a potential probiotic candidate for improving intestinal immunity, particularly in mitigating the negative consequences of antibiotic use.

TEST DB: The intelligent data management system for Toxicogenomics (독성유전체학 연구를 위한 지능적 데이터 관리 시스템)

  • Lee, Wan-Seon;Jeon, Ki-Seon;Um, Chan-Hwi;Hwang, Seung-Young;Jung, Jin-Wook;Kim, Seung-Jun;Kang, Kyung-Sun;Park, Joon-Suk;Hwang, Jae-Woong;Kang, Jong-Soo;Lee, Gyoung-Jae;Chon, Kum-Jin;Kim, Yang-Suk
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2003.10a
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    • pp.66-72
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    • 2003
  • Toxicogenomics is now emerging as one of the most important genomics application because the toxicity test based on gene expression profiles is expected more precise and efficient than current histopathological approach in pre-clinical phase. One of the challenging points in Toxicogenomics is the construction of intelligent database management system which can deal with very heterogeneous and complex data from many different experimental and information sources. Here we present a new Toxicogenomics database developed as a part of 'Toxicogenomics for Efficient Safety Test (TEST) project'. The TEST database is especially focused on the connectivity of heterogeneous data and intelligent query system which enables users to get inspiration from the complex data sets. The database deals with four kinds of information; compound information, histopathological information, gene expression information, and annotation information. Currently, TEST database has Toxicogenomics information fer 12 molecules with 4 efficacy classes; anti cancer, antibiotic, hypotension, and gastric ulcer. Users can easily access all kinds of detailed information about there compounds and simultaneously, users can also check the confidence of retrieved information by browsing the quality of experimental data and toxicity grade of gene generated from our toxicology annotation system. Intelligent query system is designed for multiple comparisons of experimental data because the comparison of experimental data according to histopathological toxicity, compounds, efficacy, and individual variation is crucial to find common genetic characteristics .Our presented system can be a good information source for the study of toxicology mechanism in the genome-wide level and also can be utilized fur the design of toxicity test chip.

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Transgenic lettuce (Lactuca sativa L.) with increased vitamin C levels using GalUR gene (GalUR 유전자를 이용한 비타민 C 증대 상추 (Lactuca sativa L.) 형질전환체 개발)

  • Lim, Mi-Young;Cho, Yi-Nam;Chae, Won-Ki;Park, Young-Soo;Min, Byung-Whan;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.115-120
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    • 2008
  • L-Ascorbic acid (vitamin C) in vegetables is an essential component of human nutrition. The objective is to transform lettuce (Lactuca sativa L.) with GalUR gene that is involved in the vitamin C biosynthesis. The cotyledons of Hwoahong (Nongwoo Bio Co.) were used to induce the callus and shoot under the selection media with MS + 30 g/L Sucrose + 0.5 mg/L BAP + 0.1 mg/L NAA + 100 mg/L kanamycin + 200 mg/L lilacillin, pH 5.2. The shoot was developed from the cut side of the explants after 3 weeks on the selection media. We successfully transformed the lettuce with GaIUR gene and analyzed the levels of vitamin C. We found that some of the lettuce transgenic lines contained higher levels of vitamin C compared with the normal one (non-transformed). Especially, some of $T_1$ lettuces inserted by GalUR showed about $3{\sim}4$ times higher content of vitamin C compared to the non-transformed lettuce. This data support the previously work performed with GLOase transgenic $T_1$ lettuces from which several times higher content of vitamin C were identified. The $T_2$ lettuces with high content of vitamin C have been selected for further analysis.

Identification of a Third Haplotype of the Sequence Linked to the Restorer-of-fertility (Rf) Gene and Its Implications for Male-Sterility Phenotypes in Peppers (Capsicum annuum L.)

  • Min, Woong-ki;Lim, Heerae;Lee, Young-Pyo;Sung, Soon-Kee;Kim, Byung-Dong;Kim, Sunggil
    • Molecules and Cells
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    • v.25 no.1
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    • pp.20-29
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    • 2008
  • Cytoplasmic male sterility (CMS), one of the most important traits in crop breeding, has been used for commercial seed production by $F_1$ hybrid cultivars of pepper (Capsicum annuum L.). To develop reliable molecular markers for allelic selection of the Restorer-of-fertility (Rf) gene, which is known to be a major determinant of pollen fertility restoration in peppers, a sequence of approximately 10 kb flanking an RAPD fragment closely linked to the Rf locus was obtained by genome walking. A homology search revealed that this sequence contained an LTR retrotransposon and a non-LTR LINE-like retrotransposon. Sequencing of this Rf-linked region to search for polymorphisms between a dominant and recessive allele revealed 98% nucleotide sequence identity between them. A third polymorphic haplotype of the Rf-linked sequence, which has 94-96% nucleotide sequence identity with the two previously isolated haplotypes, was identified among a large number of breeding lines. Utilizing polymorphic sequences in the haplotypes, PCR markers were developed for selection of particular haplotypes and used to examine the distribution of the haplotypes in diverse breeding lines, cultivars, and C. annuum germplasms. Surprisingly, the third haplotype was the predominant type in C. annuum germplasms, while its frequency in $F_1$ hybrid cultivars was relatively low. Meanwhile, analysis of breeding lines whose Rf allele genotypes and male-sterility phenotypes were already known revealed that the third haplotype was mainly present in exotic breeding lines that cause unstable male-sterility when combined with sterile cytoplasms.

Characteristics of flow field of nose-only exposure chamber for inhalation toxicity evaluation (흡입독성평가를 위한 비부노출 챔버의 유동흐름 특성)

  • Noh, Hakjae;Bong, Choonkeun;Bong, Hakyung;Kim, Yonggu;Cho, Myunghaing;Kim, Sanghwa;Kim, Daesung
    • Particle and aerosol research
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    • v.12 no.1
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    • pp.1-9
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    • 2016
  • In this work, we evaluated the characteristics of flow field and uniformity of the nose-only exposure chambers for the inhalation toxicity test. Computational fluid dynamics (CFD) modeling was carried out to demonstrate uniformity of the nose-only exposure chambers. Because it is very important in the inhalation toxicity experiments that test materials are distributed uniformly to each holder of the chamber. The test was done with these 3 types of chamber with different form to develop inhalation toxicity evaluation system, easy-to-operate system among exposure chamber used for evaluating inhalation toxicity of environmental chemical mixtures. Through CFD interpretation, nose-only exposure chamber was made with the selection of the optimal conditions. For its evaluation, one type of fragrance was selected and measured particle size distribution of each port. The gene becoming luminous to green fluorescence was combined with GPT-SPE, a type of tGFP vector, to be inhaled to the mouse. Based on this, luminous intensity was checked. As a result, total particle number concentration of each port had average value of $3.17{\times}10^6{\sharp}/cm^3$ and range of the highest and lowest concentration value was approximately ${\pm}4.8%$. Autopsy of lung tissues of mouse showed that it had clearly better delivery of gene compared to the control group.

IDENTIFICATION OF GENES EXPRESSED IN LOW-DOSE-RATE γ-IRRADIATED MOUSE WHOLE BRAIN

  • Bong, Jin Jong;Kang, Yu Mi;Choi, Seung Jin;Kim, Dong-Kwon;Lee, Kyung Mi;Kim, Hee Sun
    • Journal of Radiation Protection and Research
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    • v.38 no.4
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    • pp.166-171
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    • 2013
  • While high-dose ionizing radiation results in long term cellular cytotoxicity, chronic low-dose (<0.2 Gy) of X- or ${\gamma}$-ray irradiation can be beneficial to living organisms by inducing radiation hormesis, stimulating immune function, and adaptive responses. During chronic low-dose-rate radiation (LDR) exposure, whole body of mice is exposed to radiation, however, it remains unclear if LDR causes changes in gene expression of the whole brain. Therefore, we aim to investigate expressed genes (EGs) and signaling pathways specifically regulated by LDR-irradiation ($^{137}Cs$, a cumulative dose of 1.7 Gy for total 100 days) in the whole brain. Using microarray analysis of whole brain RNA extracts harvested from ICR and AKR/J mice after LDR-irradiation, we discovered that two mice strains displayed distinct gene regulation patterns upon LDR-irradiation. In ICR mice, genes involved in ion transport, transition metal ion transport, and developmental cell growth were turned on while, in AKR/J mice, genes involved in sensory perception, cognition, olfactory transduction, G-protein coupled receptor pathways, inflammatory response, proteolysis, and base excision repair were found to be affected by LDR. We validated LDR-sensitive EGs by qPCR and confirmed specific upregulation of S100a7a, Olfr624, and Gm4868 genes in AKR/J mice whole brain. Therefore, our data provide the first report of genetic changes regulated by LDR in the mouse whole brain, which may affect several aspects of brain function.

Anti-Aging Effect of Ligustrum japonicum Extract in the Human Fibroblast Cells (피부 섬유아세포에서 광나무 추출물의 항 노화 효과)

  • Kim, Yun-Jeong;Lee, Yu-Ri;Cheon, Jong-Woo;Lee, Hyun-Sang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.4
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    • pp.295-301
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    • 2010
  • To develop wrinkle care cosmetic ingredients, various species of plant extracts were investigated. Accordingly, Ligustrum japonicum was selected as a candidate for developing cosmetic ingredient. By high performance liquid chromatography, 31.06 % of oleanolic acid and 8.92 % of ursolic acid which are well-known for anti-wrinkle effect were analyzed. The possibility of Ligustrum japonicum fruits extract (LJE) as a cosmetic ingredient was investigated using several biomarkers related to anti-aging, including anti-wrinkle, moisturizing and anti-inflammation. Procollagen type I and hyaluronan synthase-3 gene expression were increased by LJE in a concentration-dependent manner, whereas elastase activity and matrix metalloproteinase (MMP)-1, MMP-2 and cyclooxygenase-2 gene expression were inhibited. As the results LJE is applicable for a potential cosmetic ingredient focused on anti-aging effect.

Enhancement of Transduction Efficiency and Antitumor Effects of IL-12N220L-expressing Adenovirus by Co-delivery of DOTAP

  • Youn, Je-In;Jin, Hyun-Tak;Sung, Young-Chul
    • IMMUNE NETWORK
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    • v.7 no.4
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    • pp.179-185
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    • 2007
  • Background: Adenovirus (Ad) vectors have been widely used for many gene therapy applications because of their high transduction ability and broad tropism. However, their utility for cancer gene therapy is limited by their poor transduction into cancer cells lacking the primary receptor, coxsackievirus and adenovirus receptor (CAR). Methods: To achieve CAR-independent gene transfer via Ad, we pretreated Ad with 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and analyzed their transduction efficiency into cancer cells in vitro and in vivo comparing with the virus alone. Results: Treatment of DOTAP significantly increased adenoviral gene transfer in tumor cells in vitro. Moreover, DOTAP at an optimum dose $(10{\mu}g/ml)$ enhanced IL-12 transgene expression by fivefold in tumor, and twofold in serum after intratumoral injection of adenovirus expressing IL-12N220L (Ad/IL-12N220L). In addition, cotreatment of DOTAP decreased tumor growth rate in the Ad/IL-12N220L-transduced tumor model, finally leading to enhanced survival rate. Conclusion: Our results strongly suggest that DOTAP could be of great utility for improving adenovirus-mediated cancer gene therapy.

Rapid Selection of Multiple Gene Integrant for the Production of Recombinant Hirudin in Hansenula polymorpha

  • Kim Hwa Young;Sohn Jung Hoon;Kim Chul Ho;Rao K. Jagannadha;Choi Eui Sung;Kim Myung Kuk;Rhee Sang Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.1-6
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    • 2000
  • For the rapid selection of higher recombinant hirudin producing strain in a methylotrophic yeast Hansenula polymorpha, a multiple gene integration and dose-dependent selection vector, based on a telomere-associated ARS and a bacterial aminoglycoside 3-phosphotransferase (aph) gene, was adopted. Two hirudin expression cassettes (HV1 and HV2) were constructed using the MOX promoter of H. polymorpha and the mating factor $\alpha$ secretion signal of S. cerevisiae. Multiple integrants of a transforming vector containing hirudin expression cassettes were easily selected by using an antibiotic, G418. Hirudin expression level and integrated plasmid copy number of the tested transformants increased with increasing the concentration of G418 used for selection. The expression level of HV1 was consistently higher than that of HV2 under the similar conditions, suggesting that the gene context might be quite important for the high-level gene expression in H. polymorpha. The highest hirudin producing strain selected in this study produced over 96 mg/L of biologically active hirudin in a 500-mL flask and 165 mg/L in a 5-L fermentor.

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Cloning and mulecular characterization of a nprX gene of bacillus subtilis NS15-4 encoding a neutral protease (Cloning and Molecular Characterization of a nprX gene of Bacillus subtilis NS15-4 Encoding a Neutral protease)

  • Lee, Seung-Hwan;Yoon, Ki-Hong;Nam, Hee-Sop;Oh, Tae-Kwang;Lee, Seog-Jae;Chae, Keon-Sang
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.68-73
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    • 1996
  • An nprX gene of Bacillus subtilis NS15-4 encoding a neutral protease was cloned and its molecular characteristics were analyzed. The complete nucleotide sequence indicated that there is an open reading frame (0RF) possibly encoding 521 amino acid polypeptide. The ORF used all codons expected two cysteine and a proline having a codon bias index (CBI) of 0.09 in Escherichia coli. There were homologous sequences to the consensus sequence of -35 and -10 regions of E. coli promoters and to a Shine-Dalgarno (SD) sequence located 25 bp downstream of a mojor transcription initiation site. Moreover, there were also five minor transcription initiation sites at 6. 7. 8. 14 and 15 nt downstream of the major site. Northern blot analysis revealed the presence of about 1.8 kb mRNA transcript in E. coli having the nprX gene. The nucleotide sequence was identified in GenBank to be a gene for a neutral protease of B. sutilis with six nucleotide difference in the ORF region. The flanking regions of the NprX ORF showed much more differences form those of other neutral protease genes except the nprE gene of B. subtilis, which has the most homology to the nprX gene, and of which the flanking regions were identical to those of the nprX gene.

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