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Biomedical subjects

D Y Graham

Publications and source records attributed to D Y Graham.

At least 163 records · Page 9Linked to original sources

Relationship between Helicobacter pylori iceA, cagA, and vacA status and clinical outcome: studies in four different countries.

There is continuing interest in identifying Helicobacter pylori virulence factors that might predict the risk for symptomatic clinical outcomes. It has been proposed that iceA and cagA genes are such markers and can identify patients with peptic ulcers. We compared H. pylori isolates from four countries, looking at the cagA and vacA genotypes, iceA alleles, and presentation of the infection. We used PCR to examine iceA, vacA, and cagA status of 424 H. pylori isolates obtained from patients with different clinical presentations (peptic ulcer, gastric cancer, and atrophic gastritis). The H. pylori isolates examined included 107 strains from Bogota, Colombia, 70 from Houston, Tex., 135 from Seoul, Korea, and 112 from Kyoto, Japan. The predominant genotype differed among countries: the cagA-positive iceA1 vacA s1c-m1 genotype was predominant in Japan and Korea, the cagA-positive iceA2 vacA s1b-m1 genotype was predominant in the United States, and the cagA-positive iceA2 vacA s1a-m1 genotype was predominant in Colombia. There was no association between the iceA, vacA, or cagA status and clinical outcome in patients in the countries studied. iceA status shows considerable geographic differences, and neither iceA nor combinations of iceA, vacA, and cagA were helpful in predicting the clinical presentation of an H. pylori infection.

Antigens, Bacterial↗

Relation between clinical presentation, Helicobacter pylori density, interleukin 1beta and 8 production, and cagA status.

BACKGROUND: It is not known whether cagA+ Helicobacter pylori in duodenal ulcer (DU) have enhanced virulence compared with non-DU cagA+ H pylori. AIMS: To investigate the relation between presentation, H pylori density, interleukin 1beta (IL-1beta) and IL-8 production, and cagA status. METHODS: Fifty DU and 50 gastritis patients with cagA+ H pylori and 11 with cagA- infections were studied. Bacterial density and cytokine production were assessed using the same biopsies. Cytokine production was also measured from supernatants of medium following coculture of H pylori with MKN-45 cells. RESULTS: There was no relation between H pylori density and cagA status. There was a dose dependent relation between mucosal cytokine levels and density of cagA+ H pylori. H pylori density increased to a threshold, followed by a rapid increase in cytokines and then a plateau. IL-1beta and IL-8 levels in the antrum were greater in DU than in gastritis; in the corpus the cytokine level/H pylori differed irrespective of similar H pylori densities. However, cytokine production was similar in vitro, independent of presentation or biopsy site, suggesting that host factors are critical determinants of the inflammatory response. Mucosal IL-8 and IL-1beta levels were low with cagA- and cagA+, cagE- H pylori infections. CONCLUSIONS: The increase in antral IL-1beta and IL-8 production and inflammation in DU is related to increased numbers of bacteria and not to an increase in cytokine production per cagA+ isolate. There was no evidence of enhanced virulence of H pylori from DU compared with cagA+ non-DU H pylori.

Adult↗

A reliable method for the simultaneous identification of H pylori and gastric metaplasia in the duodenum.

While an association of Helicobacter pylori infection with duodenal mucosa gastric metaplasia has been described, the details and extent of the interaction are lacking. One of the limiting factors has been the lack of a staining technique that allows simultaneous visualisation of the bacteria and gastric metaplasia in the duodenum. This report describes a new stain that allows the simultaneous visualisation of duodenal gastric metaplasia and H pylori and compares the new stain with the component stains.

Duodenum↗

Antibody against Helicobacter pylori CagA and VacA and the risk for gastric cancer.

AIM: Helicobacter pylori is associated with gastric cancer. Our aim was to investigate whether CagA or VacA seropositivity provides additional risk for gastric cancer. METHODS: Sera from 110 gastric cancer patients were sex and aged matched with asymptomatic controls. H pylori status was determined by IgG enzyme immunoassay (HM-CAP EIA); CagA status was assessed by enzyme linked immunosorbent assay (ELISA) (OraVax) and immunoblotting (Chiron), and VacA status by immunoblotting using recombinant proteins as antigens. RESULTS: H pylori infection was associated with an increased risk of gastric cancer (odds ratio (OR) = 2.19, 95% confidence interval 1.17 to 4.1). Subgroup analysis showed a significant association with intestinal type (OR = 2.94, 1.35 to 6.41), distal type (OR = 2.97, 1.39 to 6.33), early gastric cancer (OR = 3.74, 1.54 to 9.06), and age < or = 55 years (OR = 8.33, 2.04 to 34.08), but not with diffuse type (OR = 0.83), proximal type (OR = 1.0), advanced gastric cancer (OR = 1.13), or age > 55 years (OR = 1.40). Serum CagA IgG and VacA antibody positivity was present in similar proportions in patients with and without cancer, with no significant differences in histological classification, clinical stage, or location (p > 0.3). CONCLUSIONS: H pylori infection causes chronic gastritis and is associated with the development of gastric cancer. Neither CagA nor VacA seropositivity added additional information or stratification.

Adult↗

Modified Genta triple stain for identifying Helicobacter pylori.

AIM: To evaluate whether lead nitrate could replace uranyl nitrate in the Genta stain for H pylori without sacrificing the advantages of the triple stain (Steiner silver impregnation combined with Alcian blue and haematoxylin/eosin (H&E)). METHODS: A comparison was made in 16 specimens between the original triple stain and the revised version. One pathologist evaluated all sections. RESULTS: Direct substitution of lead nitrate for uranium nitrate produced well stained organisms without interfering with H&E or Alcian blue staining. No difference was found in the ability to identify bacteria in 11 cases with H pylori density of 1 or 2 (on a scale of 0 to 5). CONCLUSIONS: The potential chemical and radiological hazards associated with uranium nitrate can be eliminated by using lead nitrate without sacrificing the advantages obtained by using the triple stain.

Coloring Agents↗

Variability in the interpretation of microsatellite patterns with different electrophoretic conditions.

Microsatellite markers permit the analysis of microsatellite instability and loss of heterozygosity. Frequently, the allelotypes of microsatellites are interpreted in the presence of numerous bands in gels. The importance of different gel electrophoresis conditions in the interpretation of microsatellite patterns was tested. Microsatellite markers were used to amplify DNA from gastric cancer samples and adjacent gastric mucosa. Polymerase chain reaction (PCR) products were separated by electrophoresis through 7% polyacrylamide gels containing either 5.6 M urea and 32% formamide or 7 M urea. PCR reactions separated on urea/formamide gels resulted consistently in clear allele definition (one or two bands), whereas 7 M urea gels resulted in allele patterns that comprised multiple bands. Analysis of microsatellite abnormalities using nonformamide gels gave false negative results in just under a third of cases (four of 13). In conclusion, the interpretation of microsatellite alterations in cancer DNA is improved by using electrophoresis conditions that result in complete DNA denaturation, such as urea/formamide/acrylamide gel electrophoresis.

DNA, Neoplasm↗

Scope and consequences of peptic ulcer disease. How important is asymptomatic Helicobacter pylori infection?

H pylori infection is so common as to seem ubiquitous in many areas of the world. Transmission is believed to be primarily person to person. The pathogen invariably damages the gastric mucosa, resulting in both structural and functional abnormalities. It causes histologic gastritis and is critical in the pathogenesis of the gastritis-associated diseases, namely, gastric ulcer, duodenal ulcer, gastric adenocarcinoma, and primary gastric lymphoma. Elimination of the infection results in healing of gastritis and cure of peptic ulcer disease.

Gastroesophageal Reflux↗

Recognizing peptic ulcer disease. Keys to clinical and laboratory diagnosis.

An algorithmic approach to evaluation of dyspepsia or abdominal discomfort begins with differentiation between peptic ulcer disease and gastroesophageal reflux disease as well as recognition of alarm signs and symptoms for gastric cancer, which are indications for early endoscopy. In the absence of alarm symptoms, most patients should undergo noninvasive testing for H pylori infection with a serologic, urea breath, or stool antigen test. Factors to consider in selection of appropriate testing include reliability, specificity, sensitivity, cost, and local access and expertise. As a general rule, physicians should choose a test that has the best accuracy for the level of testing expertise available. The basic principle underlying testing for H pylori is that patients should not undergo testing unless the physician is willing to treat on the basis of a positive test result. In patients who receive treatment, confirmation of cure is important for preventing further morbidity and reducing risk of transmission of infection.

Anti-Inflammatory Agents, Non-Steroidal↗

Practical advice on eradicating Helicobacter pylori infection.

Peptic ulcer disease associated with H pylori infection is curable. The important factors in selecting therapy are efficacy of eradication, prevention of resistance, avoidance or minimization of adverse effects, patient compliance, and cost. The most effective regimens include a bismuth preparation or antisecretory drug (proton pump inhibitor or H2 receptor antagonist) plus two antibiotics administered for 14 days. Dual-drug therapies are not recommended. Triple-drug regimens are more likely to eradicate H pylori and less likely to generate resistant strains among surviving organisms. In general, cure of the infection should be confirmed 4 weeks after completion of the treatment. Antibiotic resistance is an important consideration in choosing therapy, and patients should be taught the importance of compliance. When treatment fails, antibiotic combinations should not be repeated. Considerations for anti-H pylori treatment in a managed care environment mirror those for good medical practice in general, with special attention to stringent cost-control or outcomes-driven measures.

Anti-Bacterial Agents↗

The differential diagnosis of early gastric mucosa-associated lymphoma: polymerase chain reaction and paraffin section immunophenotyping.

The distinction between benign florid lymphoid hyperplasia and low-grade gastric mucosal-associated lymphoid tissue (MALT) lymphoma may be a challenge. The presence of monoclonal B cells in Helicobacter pylori-chronic active gastritis has suggested that polymerase chain reaction (PCR) data should be viewed with caution. We investigated the reliability of PCR versus immunophenotyping in diagnosing early gastric MALT lymphoma. We studied 1511 biopsies from eight patients with high-grade primary gastric lymphoma, 25 with low-grade MALT lymphoma, 32 with atypical lymphoid infiltrates, and 39 with Helicobacter pylori-chronic active gastritis. Paraffin sections from all cases were stained with antibodies to CD20, CD3, AE1/AE3, kappa and lambda. PCR was performed on paraffin sections using the primer set VH-FR3/J(H). Using histopathology as the gold standard in diagnosis, we confirmed monoclonality in 22 of 25 MALT lymphomas (88%); a clonal band was found in 38% (15 of 39) of patients with chronic active gastritis. An immunophenotype pattern with predominance of CD20-positive cells in lymphocytic infiltrates was associated with monoclonality in 92% of cases. The presence of an enlarged irregular mantle zone was found in both monoclonal and polyclonal areas. An equal prevalence of B and T cells in lymphocytic infiltrates was associated with a polyclonal pattern in 24 of 31 cases (77%). Immunostaining of sIg (kappa and lambda) was difficult in paraffin sections and convincing proof of monoclonality was not obtained. Lymphoepithelial lesions were infrequent in gastric biopsies and their presence was highlighted with keratin stains. Because monoclonal B cells are observed in Helicobacter pylori-associated gastritis, the correct interpretation of clonality by PCR remains unclear. Paraffin section IHC using CD20 and CD3 is especially useful to confirm the diagnosis of gastric MALT lymphoma.

Adult↗

Proinflammatory activation of neutrophils and monocytes by Helicobacter pylori is not associated with cagA, vacA or picB genotypes.

Chronic Helicobacter pylori infection is associated with mucosal inflammation. The aim of the present study was to assess human neutrophil and monocyte activation induced by H. pylori strains with different virulence genotypes. Bacterial sonicates from 12 strains were used to induce phagocyte up-regulation of adherence molecule CD11b, assessed by fluorescence flow cytometry, and oxidative burst responses, assessed by chemiluminescence. A dose-dependent induction of the expression of CD11b was observed with sonicate from all H. pylori strains on both neutrophils and monocytes. Strains negative for cagA and picB genes had the same inducing activity of upregulation of CD11b as strains positive for these genes. A vacA-S2 type strain had the same activity as vacA-S1 type strains. The induction of toxic oxygen radicals by H. pylori-activated neutrophils gave higher median values for the cagA-positive strains than for the cagA-negative strains. For the monocyte chemiluminescence response, cagA-negative strains gave higher median values compared to cagA-positive strains. We conclude that upregulation of the neutrophil and monocyte adherence molecule CD11b induced by H. pylori sonicates is not associated with the presence of cagA, picB or mosaic pattern of vacA, and that cagA, picB-negative strains and vacA-S2 strains retain their inflammatory capacity.

Adult↗

Co-twin study of the effect of environment and dietary elements on acquisition of Helicobacter pylori infection.

The rate of Helicobacter pylori infection is inversely related to socioeconomic status, and childhood is thought to be the major acquisition period. The authors investigated the importance of childhood environment in the acquisition of H. pylori infection. A cross-sectional study was conducted of monozygotic and dizygotic twins who were reared apart or reared together. Three hundred twins from a subregistry of the Swedish Twin Registry were studied. H. pylori status was evaluated by using an enzyme-linked immunosorbent assay for anti-H. pylori immunoglobulin G. Socioeconomic status during childhood was assessed on the basis of the density of the living conditions and the economic situation of the family that reared the twins. Current socioeconomic status was estimated by using a scale that combined income and education. Dietary elements that were studied included fat and fiber intake and ascorbic acid consumption. The density of the childhood home was consistently found to be significantly associated with the acquisition of H. pylori infection (p = 0.04). Among monozygotic twins reared apart and discordant for H. pylori status, affected twins were raised in homes under poorer socioeconomic conditions than those of their unaffected co-twins (p = 0.02). Additionally, infected twins consumed more ascorbic acid than their unaffected co-twins (p = 0.04). The finding of an effect of socioeconomic status during childhood on the acquisition of H. pylori among monozygotic twins who were reared apart and had an identical genetic makeup but not a common environment confirms the hypothesis that childhood acquisition of H. pylori infection is linked to hygiene practices.

Adult↗

Helicobacter pylori seropositivity and coronary heart disease incidence. Atherosclerosis Risk In Communities (ARIC) Study Investigators.

BACKGROUND: Several epidemiological and clinical reports have suggested seropositivity for Helicobacter pylori may be a risk factor for coronary heart disease. However, there has been no prospective study of this association involving an ethnically diverse sample of middle-aged men and women. METHODS AND RESULTS: Using a prospective, case-cohort design, we determined H pylori seropositivity in relation to coronary heart disease incidence over a median follow-up period of 3.3 years among middle-aged men and women. There were 217 incident coronary heart disease cases and a cohort sample of 498. We determined H pylori antibody status by measuring IgG antibody to the high-molecular-weight cell-associated proteins of H pylori using a sensitive and specific ELISA. The prevalence of H pylori seropositivity was higher in blacks than whites, in those with less than high school education, in those with lower plasma pyridoxal 5'-phosphate and higher homocyst(e)ine concentrations, in those who did not use vitamin supplements, in those with higher fibrinogen levels, and in those seropositive for cytomegalovirus and herpes simplex type I (all P<0.05). The age-, sex-, race-, and field center-adjusted hazard ratio of coronary heart disease for H pylori seropositivity was 1.03 (95% CI=0.68 to 1.57). After adjustment for other risk factors, including fibrinogen, cytomegalovirus seropositivity, and herpes simplex type I seropositivity, the hazard ratio was 0.85 (95% CI=0.43 to 1.69). H pylori seropositivity also was not associated with increased mean intima-media thickness of the carotid artery, a measure of subclinical atherosclerosis. CONCLUSIONS: H pylori infection is probably not an important contributor to clinical coronary heart disease events.

Antibodies, Bacterial↗

Serological evidence of SV40 infections in HIV-infected and HIV-negative adults.

SV40 is a simian polyomavirus that was a contaminant of some viral vaccines administered to people between 1955 and 1962. SV40 DNA has recently been found associated with several types of human tumors, suggesting that the virus is present in humans. We examined sera from patients infected with human immunodeficiency virus type 1 (HIV-1) as well as from HIV-1-negative controls to determine the prevalence of SV40 neutralizing antibodies using a specific plaque reduction assay. We found that 16.1% of HIV-infected patients (n = 236) were seropositive for SV40, as compared to 12.0% of HIV-negative control volunteers (n = 108) and 11.1% of HIV-negative patients (n = 72). These differences were not statistically significant. As individuals born between 1941 and 1962 had the highest chance of having received SV40-contaminated poliovaccines, we analyzed SV40 seropositivity rates based on year of birth. SV40 antibody rates for HIV-infected patients born before 1941, between 1941 and 1962, and after 1962 were 17.1%, 16.3%, and 11.8%, respectively. For the HIV-negative subjects, the rates were 12.5%, 12.0%, and 9.7%, respectively. There was no correlation between SV40 seropositivity and either the stage of disease in HIV-infected patients or the race/ethnicity. Also, there was no correlation between the presence of SV40 neutralizing antibody and the titer of neutralizing antibody to human polyomavirus BKV. The SV40 seropositivity rates in the patients born between 1941 and 1962 may be explained by the likelihood of those individuals having received SV40-contaminated vaccines, but the detection of SV40 neutralizing antibody in individuals born after 1962 (with no risk of having received contaminated vaccines) is significant. Although cross-reactive antibodies might theoretically contribute to the observed reactivities, these results suggest that SV40 neutralizing antibodies are present in certain individuals and raise the possibility that SV40 continues to infect humans long after vaccines were freed from contamination.

AIDS-Related Opportunistic Infections↗

The effect of galE gene inactivation on lipopolysaccharide profile of Helicobacter pylori.

The galE gene product, UDP-galactose 4-epimerase, mediates the incorporation of galactose in extracellular polysaccharide materials such as the O-side chain of lipopolysaccharide (LPS). The O-side chain in H. pylori LPS has been shown to cross-react with Lewis x and/or y blood group antigens, suggesting its potential involvement in H. pylori-linked autoimmune disease. To study its role in H. pylori LPS biosynthesis, the galE gene was cloned, sequenced, and a galE-knockout H. pylori strain was constructed. The H. pylori galE gene encoded a protein of 344 amino acids with a molecular weight of 39K. The LPS profile from the galE-knockout H. pylori strain showed a lower molecular weight than that of the parental strain, indicating the involvement of the galE gene in LPS biosynthesis of H. pylori.

Amino Acid Sequence↗

Helicobacter pylori infection produces reversible glycosylation changes to gastric mucins.

The protective ability of gastric mucins may depend largely on their oligosaccharide chains. We evaluated the effects of H. pylori infection on the glycosylation of gastric mucins. Gastric biopsy specimens from 20 H. pylori-infected patients before and after cure of the H. pylori infection and 8 normal uninfected volunteers were examined by immunostaining for simple mucin-type glycoproteins and blood-group-related antigens bearing type 1 chain backbone. The immunoreactivity in different gastric compartments was evaluated. Simple mucin-type glycoproteins and blood-group-related antigens were expressed in surface mucous cells. Simple mucin-type glycoproteins showed antrum-predominant expression in normal volunteers and were found in significantly fewer surface mucous cells in infected patients than in normal volunteers; their expression was restored after eradication of H. pylori. Sialyl Lewis(a) and Lewis(b) were expressed in fewer surface mucous cells after than before eradication. The patterns of glycosylation of gastric mucins vary in different gastric compartments and are reversibly altered by H. pylori infection. These alterations may affect the protective functions of gastric mucins.

Adult↗

Antibiotic resistance in Helicobacter pylori: implications for therapy.

Helicobacter pylori is a conventional gram-negative bacteria that causes an infection of the gastric mucosa. The organism is sensitive to most common antibiotics and theoretically should be easy to eradicate, provided that the patient takes antibiotics to which the organism is sensitive using a sufficient dose and duration of therapy. Unfortunately, the infection has proved difficult to cure. There are many other examples of bacterial infections that resist treatment with antibiotics even when the individual organisms demonstrate antibiotic sensitivity in vitro (e.g., Salmonella in the biliary tract). Failure can be due to the presence of antibiotic-resistant organisms, features of the infection may make it inaccessible to the antibiotics despite having antibiotic-sensitive organisms (a resistant infection), or both. Acquired resistance of H. pylori to clarithromycin, metronidazole, tetracycline, and amoxicillin have all been described.

Biofilms↗