• Title/Summary/Keyword: H. pylori culture

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The Eradication of Helicobacter pylori in the Duodenal Ulcer in Children and the Duodenal Recurrence (소아 십이지장궤양에서의 H. pylori 박멸과 궤양재발에 대한 연구)

  • Choe, Yon-Ho;Ko, Jae-Sung;Kim, Soon-Yeong;Yoo, Young-Mee;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.1 no.1
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    • pp.30-36
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    • 1998
  • Purpose: It is well known that duodenal ulcer disease does not relapse if H. pylori is cleared from the gastric mucosa. Little is known about the recurrence of duodenal ulcer in children. The purpose of this study was to evaluate the effect of the eradication of H. pylori in duodenal ulcer in children upon the duodenal ulcer recurrence. Methods: 105 patients (M:F=78:27) diagnosed as duodenal ulcer by endoscopy in 1987~1995 were reviewed clinically, and were parted into two groups. The two treatment groups were ranitidine/antacid (RAN/ANT) and ranitidine/amoxicillin/denol (RAN/AMX/D). The latter was for H. pylori-positive children with duodenal ulcer who were diagnosed by serology and/or antral biopsies for histology, culture, and urease testing. The recurrence rates were compared between the two groups. Results: 1) 30 patients with primary duodenal ulcer underwent endoscopy for H. pylori and 27 (90.0%) of them were positive for H. pylori. 2) 27 of H. pylori-positive children received RAN/AMX/D. 23(85.2%) of them showed cure of duodenal ulcer and eradication of H. pylori. 3) The duodenal ulcer recurrence rate in RAN/ANT group was 65.3% and the rate in RAN/AMX/D was 4.3% by a year. Conclusions: There is a strong correlation between the duodenal ulceration and H. pylori infection in children, and the eradication of H. pylori in duodenal ulcer patients reduces the recurrence of the ulcer. Because of the low incidence of duodenal ulcers in children, a multicenter prospective study is required to determine the effect of treating H. pylori infetion on the long term natural history of duodenal ulcer disease.

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Real-Time PCR Detection of 16S rRNA Novel Mutations Associated with Helicobacter pylori Tetracycline Resistance in Iran

  • Dadashzadeh, Kianoosh;Milani, Morteza;Rahmati, Mohammad;Akbarzadeh, Abolfazl
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8883-8886
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    • 2014
  • Background: Tetracycline is an antibiotic widely used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing due to increasing bacterial resistance. The aim of this study was to investigate the occurrence of 16S rRNA mutations associated with resistance or reduced susceptibility to tetracycline ofHelicobacter pylori by real-time PCR (RT-PCR) assays from culture. Materials and Methods: Tetracycline susceptibility and minimal inhibition concentration (MIC) was determined by the Epsilometer test (Etest) method. A LightCycler assay developed to detect these mutations was applied to DNA extracted from culture. The 16S rRNA of these isolates was sequenced and resistance-associated mutations were identified. From 104 isolates of H. pylori examined, 11 showed resistance to tetracycline. Results: LightCycler assay was applied to DNA extracted from 11 tetracycline-susceptible and 11 tetracycline resistance H. pylori isolates. In our study the sequencing of the H. pylori wild types in 16 s rRNA gene were AGA 926-928 with MIC (0.016 to $0.5{\mu}g/ml$), while the sequencing and MIC for resistant were GGA and AGC, (0.75 to $1.5{\mu}g/ml$), respectively. Also we found a novel mutation in 2 strains with $84^{\circ}C$ as their melting temperatures and exhibition of an A939C mutation. Conclusions: We conclude that real-time PCR is an excellent method for determination of H. pylori tetracycline resistance related mutations that could be used directly on biopsy specimens.

Non-Invasive Colorimetric Magneto Loop-Mediated Isothermal Amplification (CM-LAMP) Method for Helicobacter pylori Detection

  • Bangpanwimon, Khotchawan;Mittraparp-arthorn, Pimonsri;Srinitiwarawong, Kanchana;Tansila, Natta
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.501-509
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    • 2021
  • More than half the world's population is thought to be infected with Helicobacter pylori. Although the majority of infected people are asymptomatic, H. pylori infection may cause gastric ulcers and deadly gastric cancer. Owing to the difficulty and invasiveness of current routine culture and diagnostic methods, a highly sensitive and specific noninvasive assay for H. pylori is of interest. This study highlighted the design and performance of a colorimetric magneto loop-mediated isothermal amplification (CM-LAMP) assay to detect H. pylori in spiked saliva samples. LF primers were coated on magnetic nanoparticles by carbodiimide-induced immobilization and functionally used for solid-phase amplification. During the LAMP reaction at 66℃, biotin-tagged FIPs were incorporated into LAMP amplicons. The colorimetric signal developed after the addition of NeutrAvidin horseradish peroxidase conjugate (NA-HRP) and ABTS. None of the tested microorganisms, including closely related bacteria, was shown positive by the CM-LAMP assay except H. pylori isolates. This novel platform was highly specific and 100-fold more sensitive (40 CFU/ml or 0.2 CFU per reaction) than the PCR and conventional LAMP assays for the detection of H. pylori in spiked saliva. Our results demonstrated the feasibility of using this noninvasive molecular diagnostic test to detect H. pylori in saliva samples.

Helicobacter pylori inhibited cell proliferation in human periodontal ligament fibroblasts through the Cdc25C/CDK1/cyclinB1 signaling cascade

  • Li, Huanying;Liang, Dongsheng;Hu, Naiming;Dai, Xingzhu;He, Jianing;Zhuang, Hongmin;Zhao, Wanghong
    • Journal of Periodontal and Implant Science
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    • v.49 no.3
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    • pp.138-147
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    • 2019
  • Purpose: Several studies have shown that the oral cavity is a secondary location for Helicobacter pylori colonization and that H. pylori is associated with the severity of periodontitis. This study investigated whether H. pylori had an effect on the periodontium. We established an invasion model of a standard strain of H. pylori in human periodontal ligament fibroblasts (hPDLFs), and evaluated the effects of H. pylori on cell proliferation and cell cycle progression. Methods: Different concentrations of H. pylori were used to infect hPDLFs, with 6 hours of co-culture. The multiplicity of infection in the low- and high-concentration groups was 10:1 and 100:1, respectively. The Cell Counting Kit-8 method and Ki-67 immunofluorescence were used to detect cell proliferation. Flow cytometry, quantitative real-time polymerase chain reaction, and western blots were used to detect cell cycle progression. In the high-concentration group, the invasion of H. pylori was observed by transmission electron microscopy. Results: It was found that H. pylori invaded the fibroblasts, with cytoplasmic localization. Analyses of cell proliferation and flow cytometry showed that H. pylori inhibited the proliferation of periodontal fibroblasts by causing G2 phase arrest. The inhibition of proliferation and G2 phase arrest were more obvious in the high-concentration group. In the low-concentration group, the G2 phase regulatory factors cyclin dependent kinase 1 (CDK1) and cell division cycle 25C (Cdc25C) were upregulated, while cyclin B1 was inhibited. However, in the high-concentration group, cyclin B1 was upregulated and CDK1 was inhibited. Furthermore, the deactivated states of tyrosine phosphorylation of CDK1 (CDK1-Y15) and serine phosphorylation of Cdc25C (Cdc25C-S216) were upregulated after H. pylori infection. Conclusions: In our model, H. pylori inhibited the proliferation of hPDLFs and exerted an invasive effect, causing G2 phase arrest via the Cdc25C/CDK1/cyclin B1 signaling cascade. Its inhibitory effect on proliferation was stronger in the high-concentration group.

Evaluation of the anti-Helicobacter pylori and cytotoxic properties of the antimicrobial substances from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 (Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 균주가 생산한 항균물질의 anti-Helicobacter pylori 활성 및 위장상피세포에 대한 세포독성 평가)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.156-168
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    • 2015
  • The objective of this study is to investigate the anti-Helicobacter pylori and anti-cancer activities of the live cells (LC), cell-free culture supernatants (CFCS), and bacteriocin solution (BS) obtained from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 strains. After incubation for 30 h in MRS broth, the concentration of lactic acid produced by L. paracasei BK57 ($155.9{\pm}10.2mM$) was higher than in MRS broth using L. acidophilus BK13 ($126.8{\pm}7.9mM$). Maximum bacteriocin activity (128 AU/ml) of BK13 strain was observed after 30 h of cultivation at $37^{\circ}C$, however its magnitude was significantly lower than that of BK57 strain (256 AU/ml). The LC of L. acidophilus BK13 and L. paracasei BK57 were able to inhibit the growth of H. pylori ATCC 43504 at different incubation times, depending on the initial inoculum of the LAB. These CFCS and BS obtained from BK13 and BK57 strains dramatically inhibited the growth, adhesive ability, and enzymatic activity of H. pylori. Meanwhile, the anti-cancer effect of the lactic acid from L. acidophilus BK13 and L. paracasei BK57 strains on AGS cells had significant differences with the control group. Therefore, these antagonistic substances-producing strains are potentially useful as new potential antimicrobial agents for the management and prevention of H. pylori infections.

Synergistic Inhibition of IgY, Auricularia auricula, and Lactic Acid Bacteria from Kimchi and Tarak on Helicobacter pylori (Helicobacter pylori 의 생육억제에 대한 유산균, 난황항체 및 목이버섯의 상승효과)

  • Yoo, Hye-Lim;Lee, Young-Duck;Han, Bok-Kyung;Choi, Hyuk-Joon;Park, Jong-Hyun
    • The Korean Journal of Food And Nutrition
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    • v.26 no.1
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    • pp.35-43
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    • 2013
  • The substances of lactic acid bacteria (LAB) isolated feom Kimchi and Tarak, L. mesenteriodes LAB kw5, and S. thermophilus LAB KW15 were investigated for growth effect of Helicobacter pylori with IgY and Auricularia auricula. Inhibition of H. pylori was confirmed at LAB KW5 and KW15 supernatants. Interestingly, anti-H. pylori substance in LAB KW5 and KW15 supernatants were sensitive to lipase, but insensitive to protein hydrolase and carbohydrate hydrolase. The inhibition zone toward H. pylori was not shown with the lipase-treated supernatants. Therefore, there seemed to be lipid-like substances in the cultures. By the analyses with gas chromatography, undecanoic acid ($C_{11:0}$), palmitic acid ($C_{16:0}$), stearic acid ($C_{18:0}$), and oleic acid ($C_{18:1}$) were detected at the culture substances from L. mesenteroides LAB KW5 and S. thermophilus LAB KW15, and more eicosadienoic acid ($C_{20:2}$) from L. mesenteroides LAB KW5. Anti-H. pylori substances of LAB with IgY and A. auricula extract were analyzed for inhibition effect of H. pylori. The inhibition increased more by the range from 57% to 86% by the mixture. The substances with IgY and A. auricula extract showed more effective inhibition of H. pylori than single or double trials.

HOW TO DEVELOPE NEW PRO BIOTIC WITH ANTI Helicohacter pylori FUNCTION

  • Lee Yeonhee
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2001.12a
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    • pp.161-169
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    • 2001
  • Lactic acid producing bacteria were isolated from baby feces and characterized to be used as a probiotic with anti Helicobacter pylori functions. The selected bacteria had inhibition activity on the adherance and growth of H. pylori. These bacteria had additional beneficial characteristics for the probiotic such as antibacterial activity, antitumor activity, immunostimulation activity, resistance to antibiotic and bile salt, ability to bind to the intestinal cells, and safe for the human use.

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Effect of Hot Water Extract of Alnus japonica Steud on the Experimentally-induced Acute Gastritis and Peptic Ulcers in Rats (오리나무 열수 추출물이 실험적으로 유발된 흰쥐의 위염 및 위궤양에 미치는 영향)

  • Na, Chun-Soo;Lee, Sang-Bum;Kim, Jin-Bum;Chung, Ha-Sook;Dong, Mi-Sook
    • Korean Journal of Pharmacognosy
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    • v.43 no.1
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    • pp.72-78
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    • 2012
  • Alnus japonica Steud (A. japonica) have long been used in the traditional medicine for gastric disorder, hepatitis and fatty liver in Korea. Antiulcer effects of A. japonica hot water extract (AJ ext) were evaluated by in vitro antibacterial activity against H. pylori, by the inhibitory action against the in vitro gastric $H^+/K^+$-ATPase and using rat models of gastric mucosal damage and gastric ulcer induced by HCl-ethanol, indomethacin, and restraint and water-immersion stress. For the determination of antibacterial activity of AJ ext against H. pylori, the activity of urease which released from H. pylori was measured in culture. AJ ext showed weak antibacterial activity against H. pylori with the growth inhibitions of 37% and 61% by adding final concentrations of 500 and $1000{\mu}g/ml$ culture, respectively at 24 h. To observe the inhibitory activity of AJ ext against the $H^+/K^+$-ATPase in hog gastric membrane vesicle, $IC_{50}$ value of AJ ext was $806.3{\mu}g/ml$. Pretreatment of AJ ext (200, 500 mg/kg, p.o.) prevented in a dose-dependent manner the acute gastritis in HCl-ethanol model and the formation of gastric ulcer in indomethacin model and restraint and water-immersion stress model. These results suggest that the AJ ext can be used for prevention and treatment of gastric mucosal damage and ulcers induced by various stress.

Evaluation of Factors that Can Affect Protective Immune Responses Following Oral Immunization of Recombinant Helicobacter pylori Urease Apoenzyme

  • Kim, Jang-Seoung;Chang, Ji-Hoon;Park, Eun-Jeong;Chung, Soo-Il;Yum, Jung-Sun
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.865-872
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    • 2000
  • Helocobacter phylori is the major cause of gastritis, peptic ulcer, and a principal risk factor for gastric cancer. As the firs step towards a vaccine against H. pylori infection, Hy.pylori urease was expressed and purified as a recombinant apoenzyme (rUrease) in E. coli. In order to develop an effective immunization protocol using rUrease, the host immune responses were evaluated after the oral immunization of mice with rUrease preparations plus cholera toxin relative to various conditions, such as the physical nature of the antigen, the frequency of the booster immunization, the dose of the antigen, and the route of administration. The protective efficacy was assessed using a quantitative culture following an H. pylori SS1 challenge. It was demonstrated that rUrease, due to its particulated nature, was more superior than the UreB subunit as a vaccine antigen. The oral immunization of rUrease elicited significant systemic and secretory antibody responses, and activated predominantly Th2-type cellular responses. The bacterial colonization was significantly reduced (~100-fold) in those mice immunized with three or four weekly oran doses of rUrease plus cholera toxin (p<0.05), when compared to the non-immunized/challenged controls. The protection correlated well with the elicited secretory IgA level against rUrease, and these secretory antibody responses were highly dependent on the frequency of the booster immunization, yet unaffected by the dose of the antigen (25-200$\mu\textrm{g}$). These results demonstrate the remarkable potential of rUrease as a vaccine antigen, thereby strengthening the possibility of developing an H. pylori vaccine for humans.

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Association of Helicobacter pylori cagA Gene with Gastric Cancer and Peptic Ulcer in Saudi Patients

  • Saber, Taisir;Ghonaim, Mabrouk M.;Yousef, Amany R.;Khalifa, Amany;Qurashi, Hesham Al;Shaqhan, Mohammad;Samaha, Mohammad
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1146-1153
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    • 2015
  • This study was conducted to assess the relationship between occurrence of gastric cancer and peptic ulcer, and the presence of H. pylori cagA gene and anti-CagA IgG, and to estimate the value of these antibodies in detecting infection by cagA gene-positive H. pylori strains in Saudi patients. The study included 180 patients who were subjected to upper gastrointestinal endoscopy in Taif province and Western region of Saudi Arabia (60 gastric cancer, 60 peptic ulcer, and 60 with non-ulcer dyspepsia). Gastric biopsy specimens were obtained and tested for H. pylori infection by rapid urease test and culture. PCR was performed on the isolated strains and biopsy specimens for detection of the cagA gene. Blood samples were collected and tested for CagA IgG by ELISA. H. pylori infection was detected among 72.8% of patients. The cagA gene and anti-CagA IgG were found in 63.4% and 61.8% of H. pylori-infected patients, respectively. They were significantly (p < 0.01) higher in patients with gastric cancer and peptic ulcer compared with those with non-ulcer dyspepsia. Detection of the CagA IgG was 91.6% sensitive, 89.6% specific, and 90.8% accurate compared with detection of the cagA gene. Its positive and negative predictive values were 93.8% and 86%, respectively. The study showed a significant association between the presence of the cagA gene and gastric cancer and peptic ulcer disease, and between anti-CagA IgG and the cagA gene in Saudi patients. However, a further larger study is required to confirm this finding.