• Korean Journal of Veterinary Research
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• v.46 no.4
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• pp.327-335
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• 2006

A stable and reliable Helicobacter pylori (H. pylori) infection animal model would be necessary for evaluating vaccine efficacy and helpful for understanding the pathological mechanism of the organism. The aim of the present study is to investigate the effect of ethanol treatment prior to H. pylori inoculation on associated gastric mucosal injury and to establish ethanol-pretreating animal model to study H. pylori infection. Male Mongolian gerbils were used for the study. H. pylori was orally inoculated after 12 h fasting. 3 h prior to H. pylori inoculation, a group of gerbils was orally treated with absolute ethanol, 60% and 40% ethanol respectively. Another group of animals was treated either with H. pylori culture media alone or with different concentrations of ethanol plus culture media. Gerbils were killed 4 or 8 weeks after H. pylori inoculation. The colonization of H. pylori was confirmed by both histological examination and rapid urease test. Mucosal damage was evaluated grossly and histologically according to the criteria. The colonization of H. pylori and pathological changes in gastric mucosa of the animals were also observed. Although no significant change to the gastric mucose was observed in the animals treated either with H. pylori culture media alone or with different concentrations of ethanol plus culture media, persistent H. pylori infection was seen in the mucosa and mucosal leucocyte infiltration and severe epithelial damage was observed in the Helicobacter and ethanol + Helicobacter groups after 4 weeks. The gross and histological scores were higher in the ethanol + Helicobacter than in the Helicobacter alone group. As the results, ethanol-pretreatment with 60% concentration induced severe pathogenic changes by H. pylori infection in 5 weeks-old Mongolian gerbils. These results suggested that ethanol-pretreatment before H. pylori inoculation could increase the severity of gastric mucosal inflammation and enhance the colonization of H. pylori. The established ethanol-pretreating animal model would contribute to screen new drugs against H. pylori and be used as an useful tool for various animal experiments with H. pylori strains.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.422-427
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• 2006

The culture media from Fomitopsis pinicola were extracted by methanol and examined growth inhibition against Helicobacter pylori. The culture media from 8 days fermentation of F. pinicola showed maximum inhibition activity on H. pylori in 0.25 mg as MIC value. The highest activity against H. pylori by MHCS agar diffusion medium by Fp-P1 in 22.7 mm ID among 3 peaks from methanol fraction of 8 days culture media of Fomitopsis pinicola which purified by ion-exchange chromatography. The Fp-P1 from DEAE-Sephadex A-25 have been analysed by TLC as Fp-T1, Fp-T2 and Fp-T3 by ninhydrin staining. Fp-T3 (Rf value : 0.67) was higher activity against H. pylori in 14.4 mm ID. Fp-T3 was identified as single band by HPLC and TLC. It was assumed to aminosugar by BioLC analysis and TLC staining.

• Journal of Microbiology
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• v.40 no.4
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• pp.282-288
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• 2002

Since culture of Helicobacter pylori is relatively insensitive and cumbersome, molecular detection and typing of H. pylori isolates are gaining importance for strain differentiation. In the present study genomic DNA of 42 gastric biopsies and H. pylori isolates from corresponding patients were analyzed and compared by PCR-based RFLP assay. The 1,132-bp product representing an internal portion of ureC gene of H. pylori was amplified by PCR and digested with restriction enzymes HindⅢ, AiuⅠ and PvuⅠ. The HindⅢ, AluⅠ and PvuⅠ digestion produced 4, 7, and 2 distinguishable RFLP patterns respectively from 42-H. pylori isolates. By combining all three restriction enzyme digestions, 15 RFLP patterns were observed. However, when PCR products from 42 gastric biopsy specimens were digested by restriction enzymes HindⅢ, AluⅠ and PvuⅠ, we observed 5, 8 and 2 RFLP patterns, respectively. Patterns from 34 of 42 gastric biopsy specimens matched those of corresponding H. pylori isolates from respective patients. Patterns from the remaining eight biopsy specimens differed and appeared to represent infection with two H. pylori strains. The patterns of one strain from each of these biopsies was identical to that of the isolate from corresponding patients and the second pattern presumably represented the co-infecting strain. From the study, it appears that PCR-based RFLP analysis is a useful primary tool to detect and is distinguish H. pylori strains from gastric biopsy specimens and is superior to culture techniques in the diagnosis of infection with multiple strains of H. pylori.

• Korean Journal of Food Science and Technology
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• v.36 no.5
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• pp.779-783
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• 2004

New food ingredient was developed to eradicate and protect against re-infection of Helicobacter pylori in fermentation broth of lactic acid bacteria (LAB) showing antimicrobial activity against pathogenic microorganisms such as H. pylori and Listeria monocytogenes. LAB strain CBT SL4 was identified as Pediococcus pentosaceus by 16S rDNA sequencing and its culture broth showed antimicrobial activity of 800 AU/mL against H. pylori in optimized fermentation process. Using thin layer concentration system and spray-typed fluid bed drier system, concentrated powder product showing activity of 12,800 AU/g was harvested. Product showed eradication and protection activities against H. pylori infection on feeding test (50 AU/day) using Mongolian gerbil infection model. After 4 weeks therapy of 8,000 AU/day, ${\Delta}13CO_2$ level (DOB30) decreased about 40% in urea breath test on patient with H. pylori infection. Result show concentrated culture product of P. pentosaceus CBT SL4 has eradicating and protecting activities against H. pylori infection and can be used as food-active ingredient for prevention of gastric and duodenum ulcer caused by H. pylori.

• Microbiology and Biotechnology Letters
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• v.25 no.3
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• pp.240-247
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• 1997

Bacillary is the most common form of H. pylori observed during human infection. However, it is known that the morphology change of H. pylori from bacillary to coccoid can be occurred with a response to the environmental stresses such as the nutrient depletion, accumulation of toxic metabolites, pH alteration, and exposure to antimicrobial agents. The coccoid form of H. pylori, which is viable but non-culturable in vitro, seems to be the major cause of antibiotic resistancy and high reinfectability of H. pylori. In this regard, we studied the environmental factors that can induce the morphological change in vitro of H. pylori, and the change of fatty acid composition of plasma membrane. The morphological change from bacillary to coccoid could be observed with the depletion of nutrients, pH variation and reactive oxygen species added in the culture media. This morphologic conversion was paralleled by a dramatic decrease in unsaturated fatty acids and an increase in saturated fattv acids of plasma membrane. The change in composition of membrane fatty acid seems to be a kind of protection mechanism of H. pylori against these environmental stresses.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1513-1517
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• 2006

One single lactic acid producing bacterium, isolated from kimchi, inhibited the growth and adherence of Helicobacter pylori to the human gastric epithelial cell line MKN-45. This isolate was identified as Lactobacillus plantarum and termed L. plantarum strain PL9011. The adherence of H pylori, in the presence of live or nonviable L. plantarum strain PL9011 (10-fold CFU), decreased to 14-20%. The spent culture supernatant of L. plantarum strain PL9011 resulted in the eradication of H pylori. This activity remained stable following neutralization and heat treatment, but not following pepsin treatment, thereby suggesting small peptides as the inhibitory factor. L. plantarum strain PL9011 did not produce any harmful metabolites or enzymes. The results obtained in this study suggest that the L. plantarum strain PL9011 may be a potential novel probiotic for the stomach.

• Korean Journal of Veterinary Research
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• v.47 no.1
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• pp.77-84
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• 2007

Helicobacter pylori (H. pylori) is an important bacterial pathogen that causes chronic gastritisand is associated with gastroduodenal ulcer disease, adenocarcinoma of the distal stomach, and gastricH. pylori infection associated with host agehave not been well-defined in human. To evaluate the difference in host susceptibility to infection in relationto age of acquisition of H. pylori infection, we designed an experiment involving inoculation of H. pyloriATC 43504 at different ages of Mongolian gerbils. H. pylori was inoculated at 5 weeks and 18 monthsof age, as representatives of early and late infection, respectively. Animals were sacrificed 1 week and 4weeks after challenge, and the stomach was removed from each animal for bacterial culture, histologicalexamination, and polymerase chain reaction test. 5 week-old gerbils revealed infection andmaintained continuously its infection until 4 weeks. However, old gerbils did not maintained H. pyloriinfection. These data suggest the insusceptibility of H. pylori in old Mongolian gerbils and the importanceof animal ages for successful animal experimental infection. Also, the results demonstrated that earlyinfection of H. pylori increases its host susceptibility, as compared to the case with later infection, possiblybecause of differences in host gastric mucosal factors and imunologic responses.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.3
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• pp.1635-1642
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• 2013

Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

• Biomolecules & Therapeutics
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• v.9 no.3
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• pp.162-166
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• 2001

Helicobacter pylori is a major cause of gastric-associated diseases. In our previous study, the Adhesin/CTXA2B was expressed as insoluble recombinant chimeric protein derived from the H. pylori adhesin genetically coupled to CTXA2B subunit in Escherichia coli. Since it is very important to optimize IPTG concentration, culture temperature and composition of medium to maximize cell growth and productivity, these conditional growth factors were determined for increasing the productivity of the expressed Adhesin/CTXA2B chimeric protein in Escherichia coli JM101 carrying pTEDhpa/ctxa2b. Our data demonstrate that optimal medium for increased production of chimeric protein was a YCP/Glu medium composed of 2% yeast extract, 1% casamino acid, phosphate solution [0.3% $KH_2P0_4$, 0.4% $Na_2HP0_4$, 0.25% ($NH_4)_2HPO_4$], and 0.5% glucose. In addition, optimal concentration of IPTG was 1 mM and culture temperature, $37^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.746-752
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• 2002

The purpose of the present study was to examine the antagonistic activities of Enterococcus faecalis strain PL9003 (PL9003) on Helicobacter pylori. This strain was isolated from infant feces and found to inhibit both the growth of H. pylori and its in vitro adherence to the human gastric cell line MKN-45. The binding of PL9003 to MKN-45 was observed under a light microscope after Cram staining and under a scanning electron microscope. When detected with an FITC-conjugate antibody, both viable and nonviable PL9003 were found to decrease the number of H. pylori bound to MKN-45. When detected by an enzyme-linked immunoabsorbent assay, about 70% of the H. pylori bound on MKN-45 disappeared with the four-1314 addition of viable or nonviable PL9003. The spent culture supernatant (SCS) of PL9003 also decreased the viability of H pylori even after neutralization and pepsin treatment. The above results suggest that PL9003 has a potential as a new probiotic for the stomach.