• Title/Summary/Keyword: cagE

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Chitosan Silver Nano Composites (CAgNCs) as Antibacterial Agent Against Fish Pathogenic Edwardsiella tarda (어류 병원성 균주 Edwardsiella tarda에 대한 키토산-실버 나노입자의 항박테리아 효과)

  • Dananjaya, S.H.S.;Godahewa, G.I.;Lee, Youngdeuk;Cho, Jongki;Lee, Jehee;De Zoysa, Mahanama
    • Journal of Veterinary Clinics
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    • v.31 no.6
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    • pp.502-506
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    • 2014
  • Recently nano particles have proven for wide array of bioactive properties. In the present study, antibacterial properties of chitosan silver nano composites (CAgNCs) were investigated against fish pathogenic Edwardsiella tarda. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs against E. tarda were $25{\mu}g/mL$ and $125{\mu}g/mL$, respectively. The field emission scanning electron microscope (FE-SEM) image of CAgNCs treated E. tarda showed the strongly damaged bacteria cells than non-treated bacteria. Furthermore, treatment of CAgNCs induced the level of intracellular reactive oxygen species (ROS) in E. tarda cells in concentration and time dependent manner suggesting that it may generate oxidative stress leading to bacterial cell death. In addition, MTT assay results showed that the lowest cell viability at $100{\mu}g/mL$ of CAgNCs treated E. tarda. Overall results of this study suggest that CAgNCs is a potential antibacterial agent to control pathogenic bacteria.

Investigation of Helicobacter pylori cagE Locus Diversity on Interleukin-8 Induction in AGS Cell-line (Helicobacter pylori cagE 유전자 다형성이 위상피세포주에서 Interleukin-8 유도능에 미치는 영향)

  • Lee Su-Min;Lee Hak-Sung;Lee In-Suk;Co Yu-Kyung;Han Hae-Won;Oh Jung-Hwan;Park Jae-Myung;Choi Myung-Gyu;Chung In-Sik
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.906-912
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    • 2004
  • Helicobacter pylori infection is highly prevalent, as high as 2/3 of whole population infected, in Korea. H. pylori infection initiates inflammation by induction of interleukin-8 through type IV secretion of CagA. It was recently suggested that induction failure of IL-8 is not associated with defect in cag PAI but associated with cagE locus diversity. This study was designed to investigate ability of 11-8 in-duction according to sequence variation within the cagE gene, cagA TP motifs and vacA m-types in vitro study using AGS cell-line, and to evaluate its association with different clinical outcome. Seventy-four H. pylori stains were isolated from 23 patients with gastric cancer (Ca), 24 subjects with gastritis (G) and 27 patients with duodenal ulcer (Du) in Kangnam St. Mary's Hospital, Seoul, Korea. cagE gene diversity was confirmed by the PCR-RFLP methods with MboI/NlaIII and tyrosine phosphate motifs (TPMs) of cagA was determined TPM-A and C by using DdeI/Tsp5091 restriction enzyme and TPM-B was determend by Real time PCR the method of Owen et al. and IL-8 was measured by ELISA assay. IL-8 activity was positively detected in 59 among 74 strains $(79.7\%)$. IL-8 secretion was significantly increased in MboI A and MboI B type compared to MboI C type and in MboI/NlaIII A-C and B-C type than C-C type. 1L-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. There was no significant difference in IL-8 activity among patient groups. cagE gene diversity is thought to be mainly associated with the induction of IL-8 in H. pylori infection.

Association between Genotypes and Gastric Mucosal Lymphocytes in Helicobacter pylori-infected Children (Helicobacter pylori 감염 소아에서 유전형과 위점막 림프구)

  • Yom, Hye-Won;Cho, Min-Sun;Lee, Mi-Ae;Seo, Jeong-Wan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.12 no.2
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    • pp.140-149
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    • 2009
  • Purpose: Helicobacter pylori infection is probably acquired in childhood and persists as an asymptomatic infection for decades in most individuals. It is unclear why only a minority of those infected develop a clinical manifestation, even in childhood, such as peptic ulcer disease. H. pylori infection activates local immune responses and causes lymphocyte infiltration in the gastric mucosa. We have previously reported that both T and B cells in the lamina propria play important roles in the local immune response of H. pylori-infected children. The aim of this study was to investigate the association between H. pylori genotypes and gastric mucosal lymphocytes. Methods: Twenty-five H. pylori-infected children (10 with peptic ulcer disease and 15 with gastritis) were enrolled in this study. We investigated the genotypes (cagA, cagE, vacA, and babA2) and evaluated the association with clinical manifestations, histopathology, and gastric mucosal lymphocytes. Results: The prevalence of cagA, cagE, vacA s1m1, and babA2 was 80%, 60%, 84%, and 88%, respectively. The most prevalent (68%) combination of cagA, vacA, and babA2 genotypes was cagA+/vacA s1m1+/babA2+. H. pylori genotypes were not associated with clinical manifestations, histopathology, or gastric mucosal lymphocytes. Conclusion: There was no association between the cagA, cagE, vacA, or babA2 status and gastric mucosal lymphocytes. The role of the host immune response in relation to H. pylori genotypes and disease potential in children needs further studies.

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Effects of Aromatherapy on Stress Responses, Autonomic Nervous System Activity and Blood Pressure in the Patients Undergoing Coronary Angiography: A Non-Randomized Controlled Trial (아로마요법이 관상동맥조영술 대상자의 스트레스, 자율신경계, 혈압에 미치는 효과: 비무작위 대조군설계)

  • Song, Eun Jeong;Lee, Mi Young
    • Journal of Korean Academy of Nursing
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    • v.48 no.1
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    • pp.1-11
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    • 2018
  • Purpose: The purpose of this study was to examine the effects of aromatherapy on stress responses, autonomic nervous system (ANS) activity, and blood pressure in patients hospitalized to receive coronary angiography (CAG). Methods: A non-equivalent control group with a pretest-posttest design was used. The subjects were patients admitted to the day angiography room to receive CAG at E University Hospital (34 in the experimental group and 30 in the control group). The experimental group treatment was inhalation of the aroma oil blended with lavender, ylang-ylang, and neroli at a ratio of 4:2:1 twice before and after CAG. The measurements of stress index, ANS activity, and blood pressure were performed 5 times as follows: at admission, at pre-CAG after treatment I, at post-CAG, 2 hours after treatment II, and 4 hours after treatment II. The data were analyzed using the Mann-Whitney U Test and repeated-measures analysis of variance. Results: Significant interactions in the high frequency of ANS (F=5.58, p=.005) were observed between group and time. Stress index (z=2.14, p=.016), systolic blood pressure (z=4.14, p<.005), and diastolic blood pressure (z=3.28, p=.001) were significantly different between the experimental and control groups after 4 hours of treatment II. Conclusion: The findings showed that aromatherapy was not effective before CAG, but was effective after CAG. Therefore, aromatherapy can be used as a nursing intervention for patients receiving CAG.

Effect of Recombinant CagL Immunization on the Gastric Diseases Induced by Helicobacter pylori in Mongolian gerbils (CagL 재조합 단백질 접종후에 Mongolian gerbil에서 나타나는 Helicobacter pylori 감염에 대한 반응)

  • Bak, Eun-Jung;Jang, Sung-Il;Choi, Yun-Hui;Kim, Jin-Moon;Kim, Ae-Ryun;Kim, Ji-Hye;Woo, Gye-Hyeong;Yoo, Yun-Jung;Lee, Sung-Haeng;Cha, Jeong-Heon
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.109-115
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    • 2012
  • Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

Comparative Proteome Analysis of Zerumbone-treated Helicobacter pylori (Zerumbone 처리에 따른 Helicobacter pylori의 단백질 비교분석)

  • Kim, Sa-Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.3
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    • pp.275-283
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    • 2018
  • Helicobacter pylori is a causative organism of various gastrointestinal diseases, including chronic gastritis, gastric ulcer, or gastric adenocarcinoma. Pathogenic factors, such as cytotoxin-associated protein A (CagA) and vacuolating cytotoxic protein A (VacA), play a role. This study analyzed qualitatively and quantitatively the effects of zerumbone on the changes in the protein expression levels of various H. pylori proteins, including CagA and VacA. Approximately 200 significant proteins were screened for the H. pylori 60190 (VacA positive / CagA positive; Eastern type) strain, and proteomic analysis was performed on 13 protein molecules that were clinically significant. After two-dimensional electrophoresis (2-DE), $ImageMaster^{TM}$ 2-DE Platinum software was used for quantitative measurements of protein spots. Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-TOF-MS) and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) were used for protein identification. After intensive analysis of the proteins that showed significant changes, a reverse transcription-polymerase chain reaction was performed as required to verify the results. In this study, the significance of zerumbone as a therapeutic agent for H. pylori infection was examined by screening a new pharmacological activity mechanism of zerumbone.

Virulence Genes of Helicobacter pylori in Gastritis, Peptic Ulcer and Gastric Cancer in Laos

  • Vannarath, Sengdao;Vilaichone, Ratha-Korn;Rasachak, Bouachanh;Mairiang, Pisaln;Yamaoka, Yoshio;Shiota, Seiji;Binh, Tran Thanh;Mahachai, Varocha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.9027-9031
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    • 2014
  • Background: Helicobacter pylori (H. pylori) infection is an established cause of peptic ulcers and gastric cancer. The aim of this study was to identify H. pylori genotypes and to examine their associations with geographical regions and gastritis, peptic ulcers and gastric cancer in Laos. Materials and Methods: A total of 329 Lao dyspeptic patients who underwent gastroscopy at Mahosot Hospital, Vientiane, Laos during December 2010 - March 2012 were enrolled. Two biopsy specimens (one each from the antrum and corpus) were obtained for CLO testing and only CLO test-positive gastric tissue were used to extract DNA. PCR and sequencing were identified for variants of the cagA and vacA genotypes. Results: Some 119 Laos patients (36.2%) were found to be infected with H. pylori including 83 with gastritis, 13 with gastric ulcers (GU), 20 with duodenal ulcers (DU) and 3 with gastric cancer. cagA was detected in 99.2%. East-Asian-type cagA (62%) and vacA s1c (64.7%) were predominant genotypes in Laos. vacA s1c-m1b was significantly higher in GU than gastritis (53.8% vs. 24.1%; P-value=0.04) whereas vacA s1a-m2 was significantly higher in DU than gastritis (40.0% vs. 16.9%; P-value=0.03). East-Asian-type cagA and vacA s1c were significantly higher in highland than lowland Lao (100% vs. 55.8%; P-value=0.001 and 88.2% vs. 61.5%, P-value=0.03 respectively). Conclusions: H. pylori is a common infection in Laos, as in other countries in Southeast Asia. The cagA gene was demonstrated in nearly all Laos patients, cagA and vacA genotypes being possible important factors in explaining H. pylori infection and disease outcomes in Laos.

Construction and characterization of heterozygous diploid Escherichia coli (2배체 대장균의 제조와 그 특성)

  • Jung, Hyeim;Lim, Dongbin
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.406-414
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    • 2016
  • Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

Highly Efficient Biotransformation of Astragaloside IV to Cycloastragenol by Sugar-Stimulated β-Glucosidase and β-Xylosidase from Dictyoglomus thermophilum

  • Li, Qi;Wu, Tao;Zhao, Linguo;Pei, Jianjun;Wang, Zhenzhong;Xiao, Wei
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1882-1893
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    • 2019
  • β-Glucosidases and β-xylosidases are two categories of enzymes that could cleave out non-reducing, terminal β-D-glucosyl and β-D-xylosyl residues with release of D-glucose and D-xylose, respectively. In this paper, two functional β-glucosidase Dth3 and β-xylosidase Xln-DT from Dictyoglomus thermophilum were heterologously expressed in E.coli BL21 (DE3). Dth3 and Xln-DT were relatively stable at 75℃ and were tolerant or even stimulated by glucose and xylose. Dth3 was highly tolerant to glucose with a Ki value of approximately 3 M. Meanwhile, it was not affected by xylose in high concentration. The activity of Xln-DT was stimulated 2.13-fold by 1 M glucose and 1.29-fold by 0.3 M xylose, respectively. Furthermore, the βglucosidase Dth3 and β-xylosidase Xln-DT showed excellent selectivity to cleave the outer C-6 and C-3 sugar moieties of ASI, which established an effective and green method to produce the more pharmacologically active CAG, an exclusive telomerase activator. We measured temperature, pH and dosage of enzyme using a single-factor experiment in ASI biotransformation. After optimization, the optimal reaction conditions were as follows: 75℃, pH 5.5, 1 U of Dth3 and 0.2 U of Xln-DT, respectively. Under the optimized conditions, 1 g/l ASI was transformed into 0.63 g/l CAG with a corresponding molar conversion of 94.5% within 3 h. This is the first report to use the purified thermostable and sugar-tolerant enzymes from Dictyoglomus thermophilum to hydrolyze ASI synergistically, which provides a specific, environment-friendly and cost-effective way to produce CAG.

Periodontal granulation tissue preservation in surgical periodontal disease treatment: a pilot prospective cohort study

  • Rodriguez, Jose A. Moreno;Ruiz, Antonio J. Ortiz
    • Journal of Periodontal and Implant Science
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    • v.52 no.4
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    • pp.298-311
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    • 2022
  • Purpose: The aim of this study was to evaluate the clinical outcomes of periodontal granulation tissue preservation (PGTP) in access flap periodontal surgery. Methods: Twenty patients (stage III-IV periodontitis) with 42 deep periodontal pockets that did not resolve after non-surgical treatment were consecutively recruited. Access flap periodontal surgery was modified using PGTP. The clinical periodontal parameters were evaluated at 9 months. The differences in the amount of granulation tissue width (GTw) preserved were evaluated and the influence of smoking was analyzed. Results: GTw >1 mm was observed in 97.6% of interproximal defects, and the granulation tissue extended above the bone peak in 71.4% of defects. At 9 months, probing pocket depth reduction (4.33±1.43 mm) and clinical attachment gain (CAG; 4.10±1.75 mm) were statistically significant (P<0.001). The residual probing depth was 3.2±0.89 mm. When GTw extended above the interproximal bone peak (i.e., the interproximal supra-alveolar granulation tissue thickness [iSUPRA-GT] was greater than 0 mm), a significant CAG was recorded in the supra-alveolar component (1.67±1.32 mm, P<0.001). Interproximal gingival recession (iGR) was significant (P<0.05) only in smokers, with a reduction in the interdental papillary tissue height of 0.93±0.76 mm. In non-smokers, there was no increase in the iGR when the iSUPRA-GT was >0 mm. The clinical results in smokers were significantly worse. Conclusions: PGTP was used to modify access flap periodontal surgery by preserving affected tissues with the potential for recovery. The results show that preserving periodontal granulation tissue is an effective and conservative procedure in the surgical treatment of periodontal disease.