• Title/Summary/Keyword: urease.

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Environmental Stress Response of Calcite Forming Bacteria Isolated from Concrete Pavement (콘크리트 포장도로에서 분리한 탄산칼슘형성미생물의 다양한 환경 스트레스반응)

  • Han, Sang-Hyun;Kim, Sung-Keun;Kang, Chang-Ho;Park, Joo-Young;Jeong, Jin-Hoon;So, Jae-Seong
    • KSBB Journal
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    • v.27 no.4
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    • pp.268-272
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    • 2012
  • Microbially induced calcite precipitation (MICP) has been explored for protection and consolidation of construction materials such as concrete. In this study, we isolated 54 calcite forming bacteria from concrete pavement and selected 5 isolates which showed high specific urease activity. Also response of the 5 strains against various environmental stresses was examined. BC 4 and BC 5 showed 35% and 26% viability at heat stress ($50^{\circ}C$), respectively. BC 1 and BC 4 maintained 60.4% and 70.4% viability upon osmotic stress (1 M NaCl), respectively. Among the 5 isolates BC 4 had the highest viability upon alkaline stress (pH 10).

Efficient Biotinylation of Nitrocellulose Membrane for Immuno-Filtration Capture Assay

  • Choi, Ki-Bong;Ha, Youn-Chul;Youn, Hee-Ju;Choi, Jung-Do
    • BMB Reports
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    • v.30 no.5
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    • pp.308-314
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    • 1997
  • We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

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경구투여 백신 후보물질로서의 Helicobacter pylori 외막 단백질의 조사

  • 박형배;최태부
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.129-136
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    • 1997
  • Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

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Reliability of Stool Antigen Tests: Investigation of the Diagnostic Value of a New Immunochromatographic Helicobacter pylori Approach in Dyspeptic Patients

  • Korkmaz, Huseyin;Findik, Duygu;Ugurluoglu, Ceyha;Terzi, Yuksel
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.657-660
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    • 2015
  • Background: A diagnosis of H. pylori infection can be made by invasive or non-invasive methods. Several noninvasive diagnostic tests based on the detection of H. pylori stool antigen (HpSA) have been developed. The Genx H. pylori stool antigen card test is a new rapid, non-invasive test that is based on monoclonal immunochromatographic assay. The aim of this study was to determine its sensitivity, specificity, and diagnostic accuracy for diagnosing H. pylori infection in adult patients. Materials and Methods: A total of 162 patients were included in the study. A gastric biopsy was collected for histopathology and rapid urease testing. Stool specimens for HpSA testing were also collected. Patients were considered H. pylori positive if two invasive tests (histological and rapid urease tests) were positive. Results: Using the reference test, 50.6% of the samples were positive for H. pylori infection. The Genx H. pylori antigen test was positive in 19.7% of patients. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of the Genx H. pylori antigen test were 51.6%, 96.0%, 88.8%, 76.1%, and 79.0%, respectively. Conclusions: The Genx H. pylori stool antigen card test is a new non-invasive method that is fast and simple to perform but provides less reliable results.

Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Status of Helicobacter pylori Infection among Migrant Workers in Shijiazhuang, China

  • Xia, Pu;Ma, Ming-Feng;Wang, Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1167-1170
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    • 2012
  • Background: Helicobacter pylori infection leads to many upper gastrointestinal diseases. Migrant workers are the main part of floating population in China. However, up to now, their health status has not been a focus of attention. Methods: In order to assess the status of H. pylori infection among migrant workers in Shijiazhuang, over five years we interviewed 324 individuals between 2007 and 2011. Each underwent a rapid urease test to identify H. pylori infection and socio-demographic indicators were collected using a survey questionnaire. Results: Our results showed that family income (P = 0.003), dietetic hygiene (P = 0.005), education (P = 0.004) and marital status (P = 0.007) were associated with H. pylori infection. Conclusion: We found that migrant workers had little basic knowledge of H. pylori and their prevalence of infection remains high. Therefore, we need to promote education and awareness of H. pylori and to ensure access to diagnosis and treatment for infected workers.

Development of Disposable Enzyme-linked Immunosensor Strip Platform (일회용 스트립형 효소면역센서용 플랫폼의 개발)

  • Choi, Ji-Hye;Yi, Seung-Jae;Chang, Seung-Cheol;Kim, Kyung-Chun
    • Journal of Sensor Science and Technology
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    • v.20 no.6
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    • pp.400-405
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    • 2011
  • This study introduced the development of a strip type disposable enzyme-linked immunosensor platform for the detection of IgG. Strips of the strip sensor were fabricated by using commercial nitrocellulose filter membranes and a housing holder for the strips was manufactured by using a standard injection molding process for a plastic material. An IgG-urease conjugate was prepared and used for the competitive immune-binding with sample IgG. From the enzymatic reaction between the conjugated urease and urea added, ammonia was generated and caused a localized alkaline pH change on the immobilized antibody band which was coated onto the sensor strips. This pH increase subsequently caused a color change of the antibody band in the presence of a pH indicator, phenol red. Used in conjunction with a competitive immunoassay format, the intensity of the color produced is directly linked with the concentration of target analyte, IgG, and specific measurement of IgG in a lateral flow immunoassay format was achieved over the range 100 ppb to 2000 ppb IgG.

Effect of Leweifang on HeLa Cell Vacuolation Induced by Helicobacter.pylori cytotoxin (Helicobacter pylori 독소에 의한 세포의 공포형성에 미치는 생약혼합물의 영향)

  • Kwon, Dong-Yeul;Cai, Gan;Shon, Yun-Hee;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.33 no.1 s.128
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    • pp.13-17
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    • 2002
  • Helicobacter pylori infection is associated with type B gastritis, peptic ulcer, and gastric cancer. The vacuolation of cells induced by H. pylori is thought to be essential for the initiation and maintenance of gastric infection. The roles of H. pylori cytotoxin, urease, and ammonia in the vacuolation of HeLa cells were determined. Ammonium chloride augmented the neutral red uptake induced by H. pylori toxin. Acetohydroxamic acid (AHA) failed to block the neutral red uptake induced by H. pylori toxin. Leweifang significantly prevented the vacuolation of HeLa cells induced by H. pylori toxin or H. pylori toxin and ammonium chloride. Further investigation is required to determine the mechanisms of Leweifang for the inhibition of vacuole formation of eukaryotic cells in response to the H. pylori toxin.

Anti-Helicobacter and Anti-inflammatory Effects of Sohamhyungtang in Helicobacter pylori-Infected Human Gastric Epithelial AGS cells

  • Won, SangBum;Yim, Dongsool;Choi, SungSook
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.175-182
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    • 2017
  • This study evaluated the anti-Helicobacter and anti-inflammatory effects of Sohamhyungtang (SHHT). The minimum inhibitory concentration (MIC) of SHHT against Helicobacter pylori (H. pylori) was determined by the agar dilution method. Expression of the H. pylori cagA gene in the presence of SHHT was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Inhibition of H. pylori urease by SHHT was determined by the phenol-hypochlorite assay. Antiadhesion activity of SHHT was measured by urea-phenol red reagent. Inhibition of nitric oxide (NO) production in AGS cells was measured with Griess reagent. Inducible nitric oxide synthase (iNOS) and IL-8 mRNA expression in AGS cells which were infected with H. pylori was determined by qRT-PCR. IL-8 level was measured by enzyme-linked immunosorbent assay (ELISA). The MIC of SHHT was $100{\mu}g/mL$ and the expression of cagA gene was decreased about 25 folds in the presence of SHHT. H. pylori urease was inhibited 90% by SHHT. SHHT inhibited H. pylori adhesion on AGS cell in a concentration dependent manner. mRNA expression of iNOS and IL-8 and the production of NO and IL-8 were significantly decreased in the presence of SHHT. In conclusion, SHHT showed anti-Helicobacter activity and has potent anti-inflammatory effect on H. pylori-induced inflammation in human gastric epithelial AGS cells.

Anti-bacterial and Anti-inflammatory Effects of Angelica dahurica Extracts in Helicobacter pylori-infected Human Gastric Epithelial AGS Cells (백지(Angelica dahurica) 추출물의 Helicobacter pylori에 대한 항균력 및 H. pylori로 유도한 염증반응에 대한 항염 효과)

  • Choi, Min Kyeong;Yim, Dongsool;Choi, SungSook
    • Korean Journal of Pharmacognosy
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    • v.49 no.3
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    • pp.255-261
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    • 2018
  • The aim of this study was to evaluate the anti-helicobacter activity and anti-inflammatory activity of Angelica dahurica (AD). The minimum inhibitory concentration(MIC) of AD against Helicobacter pylori(H. pylori), expression of the H. pylori cagA gene in the presence of AD was determined. Inhibition of H. pylori urease by AD, inhibition of nitric oxide (NO) production in AGS cells was measured. IL-8 mRNA expression in AGS cells which were infected with H. pylori and IL-8 level was measured. The MIC of MeOH Ex. of AD was $250{\mu}g/mL$ and the expression of cagA gene was decreased about 88% in the presence of AD. The activity of H. pylori urease was inhibited 70% by AD. mRNA expression of IL-8 and the production of NO and IL-8 were significantly decreased in the presence of AD. In conclusion, AD showed anti-Helicobacter activity and has potent anti-inflammatory effect on H. pylori-induced inflammation in human gastric epithelial AGS cells.