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  • 1
    Electronic Resource
    Electronic Resource
    Removal of Mucus for Ultrastructural Observation of the Surface of Human Gastric Epithelium Using Pronase (2002)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 7 (2002), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background. Helicobacter pylori adhering to the human gastric epithelium causes gastric diseases such as ulcer, carcinoma and lymphoma. It is thus important to observe in detail both the surface of the epithelial cells and the H. pylori that adhered to it for the elucidation of H. pylori-induced diseases by scanning electron microscopy (SEM). Since the thick mucus layer blocks the observation of the cell surface and the bacteria, it is generally eliminated during the processing for SEM by roughly mechanical methods, but these treatments also demolish the ultrastructure of the cells. We studied the nonmechanical method for removal of mucus layer of gastric epithelium using pronase.Materials and Methods. To determine the optimal concentration of pronase, mucin was used as a substrate for inhibition of the viscosity. Pronase was added in 2% mucin at the concentration of 10, 50, 100, 500, 1000, 2000 or 5000 unit/ml and the flowing time of the mixture was measured. Based on the digestion experiment, biopsied specimens from 24 patients with dyspepsic symptoms were fixed in glutaraldehyde and then washed in rolling with different concentration of pronase. After the pretreatment by pronase, the specimens were treated according to the standard process for SEM.Results. We succeeded in removing the mucus layer on the surface of epithelial cells from the biopsied specimens fixed in glutaraldehyde by rinsing with 2000 unit/ml pronase for 24 hours.Conclusions. Using our digestive method without destroying the ultrastructure, the earliest stage which H. pylori has adhered onto the human gastric epithelium can be observed for the investigation of H. pylori-induced gastric disorders by SEM.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1083-4389.2002.00070.x
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  • 2
    Electronic Resource
    Electronic Resource
    Decreased Adherence of cagG-DeletedHelicobacter pylori to Gastric Epithelial Cells in Japanese Clinical Isolates (2002)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 7 (2002), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background. The cag pathogenicity island (cag PAI) is a major virulence factor. The ability of Helicobacter pylori to adhere to gastric epithelial cells is an important initial step for virulence. The aim of this study was to evaluate the relationship between genetic variations of cag PAI in Japanese clinical isolates and the ability of H. pylori to adhere to gastric epithelial cells.Materials and Methods. The polymerase chain reaction and Southern blot analysis were used to verify the presence or absence of cagA, cagE, cagG, cagI and cagM in the cag PAI in 236 Japanese clinical isolates. The ability of H. pylori to adhere to KATOIII cells was examined by flow cytometry.Results. Seven (3.0%) cag PAI partial-deleted strains were found in 236 clinical isolates, and these strains showed three patterns in the deleted region within the cag PAI. All of the cagG-deleted strains showed decreased adherence to KATOIII cells, in comparison with cagG-positive strains. These strains had abolished IL-8 induction despite the presence of cagE, which is essential for IL-8 induction.Conclusions. Our results suggest that cagG or surrounding genes in the cag PAI has a function related to adhesion to epithelial cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.2002.00052.x
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  • 3
    Electronic Resource
    Electronic Resource
    Guidelines in the Management of Helicobacter pylori Infection in Japan (2001)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 6 (2001), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In preparation of the approval of Helicobacter pylori therapy by the Japanese national health system, the board of directors of the Japanese Society for Helicobacter Research decided to prepare guidelines on the diagnosis and treatment of H. pylori infection for physicians in routine medical practice.〈section xml:id="abs1-3"〉〈title type="main"〉Methods.A guidelines preparation committee was formed and six meetings were held. Then, in December 1999, a consensus meeting was held in Kobe to obtain the opinions of general practitioners as well as experts from Europe, North America, and Asia.〈section xml:id="abs1-4"〉〈title type="main"〉Results. Helicobacter pylori eradication therapy is recommended in gastric or duodenal ulcer patients. Helicobacter pylori eradication therapy is recommended or gastric mucosa associated lymphoid tissue (MALT) lymphoma but it should be done at specialist institutions. The significance of H. pylori eradication therapy is still under evaluation in patients with hyperplastic polyps, chronic atrophic gastritis, non-ulcer dyspepsia and in patients after endoscopic mucosal resection of gastric cancer and after gastrectomy for gastric cancer. When diagnosing H. pylori infection, at least one of the tests requiring endoscopic biopsy (e.g. rapid urease test, histology, or culture) and tests not requiring biopsy (e.g. measurement of H. pylori antibody or urea breath test) should be used. Multiple tests are recommended to increase the accuracy. The drugs of first choice currently covered by the national health insurance system in Japan are: lansoprazole (30 mg) 1 capsule twice daily, amoxicillin (250 mg) 3 capsules twice daily, and clarithromycin (200 mg) 1–2 tablets twice daily. These three drugs should be administered after breakfast and dinner for 1 week.〈section xml:id="abs1-5"〉〈title type="main"〉Conclusion.These guidelines are intended for utilization in routine medical practice after the Japanese national health system begins to cover the management of H. pylori infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.2001.00027.x
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  • 4
    Electronic Resource
    Electronic Resource
    Can Proton Pump Inhibitors Be Used as Bactericidal Drugs in Helicobacter pylori–Positive Patients? (1999)
    Boston, MA, USA : Blackwell Science Inc
    Helicobacter 4 (1999), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.1999.99070.x
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  • 5
    Electronic Resource
    Electronic Resource
    Clinical Usefulness of Urine-based Enzyme-linked Immunosorbent Assay for Detection of Antibody to Helicobacter pylori: A Collaborative Study in Nine Medical Institutions in Japan (2000)
    Boston, MA, USA : Blackwell Science Ltd
    Helicobacter 5 (2000), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background. A urine-based enzyme-linked immunosorbent assay (ELISA) kit for detection of antibody to Helicobacter pylori has been developed in Japan. Urine samples can be obtained noninvasively and are easier and safer to handle than are serum samples. The aim of this study was to examine the clinical usefulness of this urine-based ELISA kit.Materials and Methods. A pair of random, single-void urine and serum samples was collected from each of 1,061 subjects, including 238 patients with gastroduodenal disease. The sensitivity and specificity of the urine-based ELISA was compared with those of three commercially available serum-based ELISA kits. For those patients with gastroduodenal disease, the urine- and serum-based ELISA results were also compared with those for other diagnostic methods using endoscopic biopsy specimens, such as culture, histology, and rapid urease tests.Results. Based on the three serum-based ELISA results, the sensitivity, specificity, and accuracy of the urine-based ELISA were 97.7%, 95.6%, and 96.8%, respectively. On the basis of the biopsy test results, the sensitivity (96.2%), specificity (78.9%), and accuracy (91.0%) of the urine-based ELISA were almost equivalent or superior to all three serum-based ELISAs tested. In addition, 10 of the 12 false-positive cases for urine-based ELISA were confirmed to be true positives for antibodies to H. pylori by Western blot analysis and inhibition ELISA.Conclusions. The urine-based ELISA (URINELISA H. pylori Antibody) is very accurate and should be useful as an alternative to serum-based ELISAs for screening of H. pylori infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.2000.00017.x
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  • 6
    Electronic Resource
    Electronic Resource
    Highly Sensitive Urine-Based Enzyme-Linked Immunosorbent Assay for Detection of Antibody to Helicobacter pylori (1998)
    Cambridge, MA, USA : Blackwell Publishing Ltd
    Helicobacter 3 (1998), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Enzyme-linked immunosorbent assay (ELISA) has been widely used for detection of Helicobacter pylori (H. pylori), but sample collection is often invasive, complicated, and expensive. Urine samples can be obtained noninvasively and are easier and safer to handle than serum samples. A urine-based ELISA, if found to be accurate, would therefore be a useful alternative to serum-based tests for H. pylori.〈section xml:id="abs1-2"〉〈title type="main"〉Methods.An ELISA method was developed for detection of antibodies to H. pylori in urine. Its sensitivity and specificity were compared with those of three commercially available serum-based ELISA kits and the 13C urea breath test (13C-UBT) using samples from 99 healthy volunteers and 20 patients with gastric disorders.〈section xml:id="abs1-3"〉〈title type="main"〉Results.With the assumption that 13C-UBT results are 100% accurate, the sensitivity and specificity of the urinary ELISA were 99% and 100%, respectively, and the accuracy (99%) was superior to those of the three serum ELISAs tested. Immunostaining profiles on Western blot analysis using serum samples were almost identical to those obtained using paired urine samples.〈section xml:id="abs1-4"〉〈title type="main"〉Conclusions.These findings suggest that the differences observed among ELISA test results may be due principally to differences between the profiles of antigen coated on plates for the assays, rather than to differences between antibodies in serum and urine. The urine-based ELISA (URINELISA H. pylori) developed in this study is very accurate and would be useful for screening H. pylori infection as an alternative to serum ELISAs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.1998.08045.x
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  • 7
    Electronic Resource
    Electronic Resource
    Serum Antibodies to Helicobacter pylori and its Heat-Shock Protein 60 Correlate with the Response of Gastric Mucosa-Associated Lymphoid Tissue Lymphoma to Eradication of H. pylori (2004)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 9 (2004), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background and aims.  Eradication of Helicobacter pylori leads to regression of mucosa-associated lymphoid tissue (MALT) lymphomas. In this study, we measured serum antibodies to H. pylori and H. pylori-recombinant heat-shock protein 60 (rHSP60) in patients with gastric MALT lymphoma to determine whether humoral immune responses to the bacterial antigens correlate with the efficacy of eradication therapy.Methods.  Serum samples were obtained from 33 patients with H. pylori-positive gastric MALT lymphoma before undergoing therapy to eradicate the bacteria. Anti-H. pylori antibodies were measured in a commercial assay and in immunoassays to lysates and rHSP60 which were prepared from ATCC 43504 strain.Results.  Helicobacter pylori were eradicated in all 33 patients, and the lymphoma completely regressed histologically in 26 patients (79%). Pre-treatment titers of serum antibody to H. pylori and to rHSP60 in the patients whose tumor regressed were significantly higher than titers in patients whose tumors did not regress (p = .0011 and .035, respectively). By logistic regression analysis, age (odds ratio = 0.88, 95% confidence interval = 0.80–0.99), endoscopic appearance (0.053, 0.004–0.65), titers of anti-H. pylori antibodies (67.6, 2.5–1800), and titers of anti-rHSP60 antibody (6.4, 1.2–36) were identified as significantly associated factors with the outcome of MALT lymphoma.Conclusions.  Measurement of serum antibodies to H. pylori and HSP60 might be useful for predicting the response of gastric MALT lymphoma to eradication of H. pylori.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1083-4389.2004.00225.x
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  • 8
    Electronic Resource
    Electronic Resource
    A Modified McMullen's Staining for Helicobacter pylori: A High-Contrast, Visibly Prominent Method (1998)
    Cambridge, MA, USA : Blackwell Science Ltd
    Helicobacter 3 (1998), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Many stains can be used for the histological assessment of Helicobacter pylori, but all have defects from the standpoint of sensitivity, convenience, stability, visible prominence and cost. This paper describes a stain which produces satisfactory results on all the above accounts.〈section xml:id="abs1-2"〉〈title type="main"〉Materials and Methods.Paraffin-embedded sections were dewaxed and treated with 0.2% w/v carbol fuchsin for two min; then 1.0% w/v malachite green was applied for two minutes.〈section xml:id="abs1-3"〉〈title type="main"〉Results.This technique stained the bacterial form a deep magenta against a light green background. The characteristic morphology of H. pylori was identified very easily as a stumpy curved rod, and the coccoid forms were also identified. Staining was not specific with mixed samples of bacteria, but it was sufficient to identify H. pylori in the gastric mucosa.〈section xml:id="abs1-4"〉〈title type="main"〉Conclusion.We have established an easy, stable, convenient, low-cost, and easily recognizable staining method for H. pylori in paraffin-embedded tissue.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.1998.08013.x
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  • 9
    Electronic Resource
    Electronic Resource
    Atrophic Gastritis and Intestinal Metaplasia in Japan: Results of a Large Multicenter Study (2001)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 6 (2001), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: This study evaluated the relationship between Helicobacter pylori infection, atrophic gastritis and intestinal metaplasia in Japan.〈section xml:id="abs1-3"〉〈title type="main"〉Materials and Methods.This was a multicenter study performed in 21 centers in Japan. A total of 2455 individuals were enrolled. H. pylori status was determined by validated ELISAs. Atrophic gastritis was diagnosed by histology, endoscopy with Congo Red dye scattering or the Kimura-Takemoto endoscopic classification.〈section xml:id="abs1-4"〉〈title type="main"〉Results.Atrophic gastritis increased from 9.4% in those less than 20 years of age to 〉 70% in those aged 60 or older and was strongly associated with H. pylori infection. The overall prevalence of atrophic gastritis in H. pylori infection was 82.9% (1272/1534) compared with 9.8% (90/921) among uninfected (OR = 44.8; 95% CI = 34.7–57.8). Intestinal metaplasia was present in 43.1% (542/1258) of H. pylori positive persons compared with 6.2% (51/823) among the uninfected (OR = 11.5; 95% CI = 8.5–15.5). Atrophic gastritis in H. pylori positive Japanese was very high in the younger generation (38.5% in those aged 20 or less and 58.5% in those 21–30).〈section xml:id="abs1-5"〉〈title type="main"〉Conclusions.Atrophic gastritis and intestinal metaplasia were strongly associated with H. pylori and not with aging. The fall in prevalence of H. pylori in Japan has not been associated with a corresponding fall in the prevalence of atrophic gastritis among those with H. pylori infection. The high prevalence of the precursor lesion, atrophic gastritis with intestinal metaplasia, among those with H. pylori infection suggests that the risk of development of early gastric cancer will continue to remain high in Japan.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.2001.00042.x
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  • 10
    Electronic Resource
    Electronic Resource
    Molecular Cloning of p53 cDNA of Mongolian Gerbil and Establishment of Yeast p53 Functional Assay System (2003)
    Oxford, UK : Blackwell Science Ltd
    Helicobacter 8 (2003), S. 0 
    Details
    ISSN: 1523-5378
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background. Epidemiological studies have shown a correlation between Helicobacter pylori infection and human gastric carcinogenesis. A Mongolian gerbil model has demonstrated that H. pylori infection induced gastric carcinoma. However, the disadvantage of this animal model is a lack of information regarding the cellular genes involved in oncogenesis. Mutation of the p53 gene is one of the most common steps in gastric carcinogenesis. In this study, we aimed to clone the p53 gene of the Mongolian gerbil and detect the functional mutations in H. pylori-infected animals.Materials and Methods. The p53 complementary DNA (cDNA) of Mongolian gerbil was cloned by the methods of reverse-transcribed polymerase chain reaction and rapid amplification of cDNA ends.Results. The p53 cDNA of Mongolian gerbil has a 78.8% homology to that of humans. A novel yeast p53 assay system was established and enabled to detect the functional mutations of the p53 gene in the stomach of the Mongolian gerbil.Conclusions. This is the first report of the complete sequence of wild-type p53 cDNA of the Mongolian gerbil. This genetic information and an assay system designed to detect the functional mutations of the p53 gene are useful for further investigations of gastric oncogenesis in this animal model.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1523-5378.2003.00127.x
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