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D Y Graham

Publications and source records attributed to D Y Graham.

At least 145 records · Page 8Linked to original sources
PubMed1999

Different penicillin-binding protein profiles in amoxicillin-resistant Helicobacter pylori.

BACKGROUND: The beta-lactam group of antibiotics kills bacteria by inhibiting the terminal stages of peptidoglycan metabolism. We have recently identified amoxicillin-resistant Helicobacter pylori, none of which expressed beta-lactamase. Penicillin-binding proteins (PBPs) represent a group of target enzymes for the beta-lactam antibiotic family, and alterations in PBPs have been described in other penicillin-resistant bacteria. The amoxicillin-resistant phenotype characteristically was lost after freezing but could be restored by consecutive transfers into gradient plates. MATERIALS AND METHODS: To determine whether amoxicillin resistance in H. pylori was related to alterations in any of the H. pylori PBPs, five H. pylori strains resistant to amoxicillin and three amoxicillin-sensitive strains were tested. PBPs were extracted from bacteria grown to logarithmic phase, labeled in vivo with 3H-benzylpenicillin, and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. Four main PBPs were separated from all amoxicillin-sensitive H. pylori strains. RESULTS: Only three of the four main PBPs were found in the amoxicillin-resistant H. pylori strains. The differentially detectable PBP (PBP D) had an apparent molecular weight of 30 to 32 kD. CONCLUSION: These results suggest that PBP D might play a role in the amoxicillin-resistant phenotype of H. pylori strains lacking beta-lactamase activity.

Amoxicillin↗
PubMed1999

Relationship between the cagA 3' repeat region of Helicobacter pylori, gastric histology, and susceptibility to low pH.

BACKGROUND & AIMS: The variation in size of Helicobacter pylori CagA is related to repeat sequences in the 3' region of the cagA gene. We investigated whether structural subtypes of the cagA 3' region are associated with presentation of the infection or to susceptibility to acid. METHODS: We examined 319 cagA-positive H. pylori isolates: 84 isolates from Bogota, Colombia; 83 from Houston, Texas; 24 from Siena, Italy; and 128 from Seoul, Korea. The cagA 3' region was amplified by polymerase chain reaction. Gastric histology and susceptibility to pH 3 were evaluated in relation to the number of cagA repeat regions. RESULTS: Strains with more than three repeat regions were associated with significantly higher scores for gastric mucosal atrophy and intestinal metaplasia than those with fewer repeat regions. H. pylori strains with three repeat regions were also significantly more susceptible to pH 3 than isolates with fewer repeat regions. CONCLUSIONS: H. pylori strains with more than three repeat regions in the 3' region of the cagA gene are associated with enhanced histological injury and with reduced survival in acidic conditions. It is hypothesized that these variants arise within the stomach.

Adult↗
PubMed1999

Ulcer and gastritis.

As in previous years, developments in the field of ulcers and gastritis have been dominated by new findings related to Helicobacter pylori. With the decrease in the frequency of H. pylori infection, the relative proportion of non-H. pylori ulcers has increased. Attempts to reduce the endoscopy workload by H. pylori or CagA screening have not been successful, and are probably ill-advised. It has become increasingly clear that curing H. pylori infection will not automatically lead to complete relief of symptoms in patients with duodenal ulcer disease. Post-therapy confirmation of cure will probably become the norm. Studies comparing omeprazole to misoprostol or ranitidine for nonsteroidal anti-inflammatory drug (NSAID) ulcer prevention in true NSAID ulcers have shown that omeprazole is equal to full-dose misoprostol for ulcer healing and to the lowest useful dose of misoprostol for ulcer prevention. H2-receptor antagonists cannot be recommended for NSAID ulcer healing or prevention. Elimination of H. pylori increases the prevalence of gastroesophageal reflux disease in a population in such a way that superficially, there appears to be a choice between more gastroesophageal reflux disease or multifocal atrophic gastritis. The risk of developing adenocarcinoma of the esophagogastric junction is many times (10-fold to 60-fold) less than the risk of developing gastric cancer from CagA-positive H. pylori infection with multifocal atrophic gastritis - the "protective" lesion.

Adult↗
PubMed1999

Natural history of Helicobacter pylori infection in childhood: 12-year follow-up cohort study in a biracial community.

We assessed the pattern of acquisition and loss of Helicobacter pylori infection in a cohort of 212 children from a biracial community with a homogeneous socioeconomic class. The children were followed over 12 years (1973-1974 to 1985-1986) from childhood to young adulthood. H. pylori status was assessed by the presence of serum IgG antibodies to H. pylori. At ages 7-9, 19% of children had H. pylori infection (40% of blacks vs. 11% of whites; P = .0001); 12 years later, 33% were seropositive. The higher prevalence among blacks remained (P = .0001). During follow-up, 22% of children became infected; the rate of acquisition was fourfold greater among blacks than among whites (P = .001). Over the 12-year period, infection was lost in 50% of whites compared with 4% of blacks who either remained infected or became reinfected. H. pylori infection in childhood is affected by both acquisition and loss of infection in different ethnic groups. This observation is critical for understanding the epidemiology and transmission of H. pylori infection.

Adult↗
PubMed1999

Amoxycillin tolerance in Helicobacter pylori.

Resistance to amoxycillin in Helicobacter pylori has only recently been reported. To demonstrate the existence of resistance, and to test for the presence of tolerance, 17 amoxycillin-resistant strains of H. pylori, first isolated in Sardinia (Italy) and the USA, were studied. Four amoxycillin-sensitive strains were used as controls. Primary isolates of all test strains exhibited amoxycillin resistance; beta-lactamase activity was not detected. Amoxycillin resistance was lost after storage of strains at -80 degrees C but could be rescued by plating these strains on to amoxycillin gradient plates. MICs and MBCs from rescued isolates ranged from 0.5 to 32 mg/L and from 32 to > 1024 mg/L, respectively. MBC/MIC ratios > or = 32 are characteristic of antibiotic tolerance. The ratios of MBC/MIC of amoxycillin ranged from 32 to > 1024 for the test strains, indicating that these strains were tolerant to the antibiotic. Amoxycillin resistance does occur in H. pylori. Amoxycillin susceptibility testing of H. pylori isolates in patients who fail therapy should include determination of the MBC to detect tolerance.

Amoxicillin↗
PubMed1999

Normalizing results of 13C-urea breath testing for CO2 production rates in children.

BACKGROUND: The 13C-urea breath test detects the presence of Helicobacter pylori from an enrichment of breath 13CO2, which, in turn, is critically dependent on the amount of dilution by endogenous CO2 production. The production of CO2 differs according to age (adults > children), sex (male > female) weight, and height. The cutoff value of 2.4 delta%(delta over baseline, DOB) for the 13C-urea breath test, defined in adults, does not take into account actual CO2 production. Therefore, this cutoff value (2.4 delta%) may or may not be appropriate for children. The purpose of this study was to determine a cutoff value that would provide accurate results in pediatric patients, independent of their differences in anthropometric parameters. METHODS: Estimates of CO2 production were combined with DOB values to calculate the host-dependent urea hydrolysis rate. RESULTS: Calculated as urea hydrolysis rate, the cutoff range for adults was 10.4 to 10.9 microg/min. Individual ranges were concentric (men, 9.6-10.9 microg/min; women, 8.5-12.2 microg/min). Results in studies of 312 children show that a urea hydrolysis rate of more than 10 m microg/min may also be appropriate to predict H. pylori infection. CONCLUSION: Calculating 13C-urea breath test values as urea hydrolysis rate removes the effect of individual anthropometric differences on test outcome and provides a single cutoff value for pediatric patients of all ages.

Adolescent↗
PubMed1999

Costs of diagnosis and treatment of Helicobacter pylori infection: when does choosing the treatment regimen based on susceptibility testing become cost effective?

OBJECTIVE: Antibiotic-resistant Helicobacter pylori (H. pylori) strains are becoming increasingly prevalent. Currently, most physicians treat H. pylori infections without relying on antimicrobial susceptibility testing to choose the best regimen. This study was conducted to evaluate whether routine pretreatment susceptibility testing is cost effective from a third party payer point of view. METHODS: A decision model was devised to compare direct costs and outcome for diagnosis and treatment over 1 year for two different strategies. Strategy A: Endoscopy plus biopsy followed by an empirical antibiotic treatment of H. pylori-positive ulcer patients. Treatment failure was followed by reendoscopy with biopsy and antibiotic susceptibility testing and a second treatment. Strategy B: Endoscopy as in strategy A now followed by antibiotic susceptibility testing and tailored antibiotic treatment. Treatment failure was handled as in strategy A. RESULTS: Following through with strategy A or B, the overall cure rate for both strategies was virtually identical. Therefore, cost effectiveness is defined as money saved per patient by using strategy B, while achieving similar effectiveness (cure rates). As an example we compared therapies for a population with known parameters for antibiotic resistance as well as cure rates. Pretreatment susceptibility testing would save $37,000 per 1,000 patients treated. According to our model (equal therapy-price assumption in strategy A and B), pretreatment susceptibility testing for metronidazole is less costly for all the reported populations worldwide. CONCLUSIONS: Our decision analysis suggests that routine pretreatment susceptibility testing can be cost effective under various settings. The model presented is easily transferable to any population as long as the following variables are known: 1) the proportion of H. pylori strains in the population that are resistant to the antibiotics of the initial regimen; 2) the cure rate in sensitive H. pylori strains; 3) the cure rate in resistant H. pylori strains; and 4) the costs for diagnosis and treatment used.

Biopsy↗
PubMed1999

Garlic or jalapeño peppers for treatment of Helicobacter pylori infection.

OBJECTIVE: There have been a number of reports that natural foods such as garlic, honey, and capsaicin can inhibit Helicobacter pylori in vitro and each report has suggested the natural ingredient be used for treatment of the infection. We investigated whether garlic or capsaicin-containing peppers would actually inhibit H. pylori in vivo. METHODS: We performed a prospective crossover study in healthy H. pylori-infected adults. We used the urea breath test to assess the status of the H. pylori infection. On separate days subjects received three test meals consisting of beef, tortillas, and salad with one of the following: fresh garlic (10 sliced cloves), capsaicin (six sliced fresh jalapeños), two tablets of bismuth subsalicylate (Pepto-Bismol, positive control), or nothing added (negative control). Breath testing was done before the first meal, the evening meal, and the following morning. At least 2 days elapsed between the test substances. RESULTS: Twelve subjects participated (seven men, five women), with an average age of 41.4 yr, range 27-51 yr. Ten subjects received garlic, six received jalapeños, and 11 received bismuth. Neither garlic nor capsaicin had any in vivo effect on H. pylori (median urease activity 28.5 vs 39.8 and 43.7 vs 46.6 before and after garlic and jalapeños, respectively) (p > 0.8). Bismuth had a marked inhibitory effect (median 55.8 vs 14.3 before and after bismuth) (p < 0.001), respectively. CONCLUSIONS: This study did not support a role for either garlic or jalapeños in the treatment of H. pylori infection. Caution must be used when attempting to extrapolate data from in vitro studies to the in vivo condition.

Adult↗
PubMed1999

Citric acid as the test meal for the 13C-urea breath test.

OBJECTIVE: Test meals are used in the urea breath test to slow gastric emptying and to increase the area of contact with the substrate. Recently, citric acid has been suggested as an improved liquid test meal. The mechanism is unknown and could act by delaying gastric emptying, decreasing the pH at the site of the bacteria, or both. Our aim was to evaluate the effects of citric acid test meals on urea hydrolysis in vivo, to identify the possible mechanism for enhanced urea hydrolysis, and to identify the minimum effective dose. METHODS: We compared the U.S. commercial 13C-urea breath test with four liquid test meals (200 ml of water) consisting of citric acid, ascorbic acid, sodium citrate, and glucose polymer and also after the subcutaneous administration of pentagastrin. We studied healthy volunteers with and without proven H. pylori infection (by serology and histology). 13C-urea was administered orally simultaneously with the liquid test meals or immediately after the pudding had been ingested. Breath samples were taken before and after oral administration of the 13C-urea. RESULTS: A dose response in urease activity was evident as the amount of citric acid was increased from 1 to 4 g. Citric acid at 1, 2, or 4 g produced significant increases in breath 13CO2 activity, compared with the commercial pudding (p < 0.05). Ascorbic acid (p = 0.053), subcutaneous pentagastrin (to lower pH) (p = 0.199), and glucose polymer (p = 0.03) (to delay gastric emptying) all approximately doubled breath 13CO2, compared with the commercial kit. Nevertheless, the increases were all significantly less than with the 4 g citric acid test meal. CONCLUSIONS: The data are consistent with the marked effect of citric acid on gastric emptying and, possibly, distribution of the urea within the stomach being largely responsible for the enhanced urease activity with citric acid test meals. It should be possible to use a low dose of citric acid (e.g., 1 g per 200 ml) to enhance the simplicity and palatability of the test.

Ascorbic Acid↗
PubMed1999

Genotypes of Helicobacter pylori obtained from gastric ulcer patients taking or not taking NSAIDs.

OBJECTIVE: Whether Helicobacter pylori infection and use of nonsteroidal antiinflammatory drugs (NSAIDs) are independent risk factors for ulcerogenesis remains unclear. We undertook this study to evaluate H. pylori isolates from gastric ulcer patients to determine whether the genotype of the infecting isolate could be correlated with the use or nonuse of NSAIDs. METHODS: Fifty-two patients presenting with gastric ulcer and infected with H. pylori were included; 26 patients were taking NSAIDs or aspirin (ASA) regularly at the time of ulcer diagnosis. Polymerase chain reaction (PCR) was employed to assess the presence and mosaicism of the following H. pylori genes: cagA, vacA, iceA, and picB. RESULTS: We found no statistical differences in the presence of these genes in H. pylori isolates from gastric ulcer patients taking or not taking prescription NSAIDs or ASA. A 297-bp fragment of the cagA gene was detected in 96% of the isolates from the NSAID and ASA users and 100% from the non-NSAID users (p = 1.0). A larger and more variable region of the cagA gene was detected more frequently among the isolates from non-NSAID users than those from NSAID users (p = 0.05). Ninety-two percent of the isolates were identified as vacA genotype s1. The dominant vacA subtype was s1b, 76.9% and 65.4% in isolates from non-NSAID-taking or NSAID-taking patients, respectively (p = 0.4). iceA1 genotype was not correlated with gastric ulcer as this allele was only detected in 17.3% of all isolates. CONCLUSIONS: No significant differences in the presence of the candidate virulence genes vacA, cagA, picB, or iceA were detected in isolates from gastric ulcer patients taking prescription NSAIDs or ASA, compared with those not taking these drugs, indicating that single gene presence does not allow discrimination of isolates that may be important in NSAID-induced ulcerogenesis. A variable region of the cagA gene was more frequently detected in isolates from patients not taking NSAIDs or ASA, suggesting that this gene may be modified by NSAID- or ASA-related factors or that certain strains may be selected for in patients taking these medications.

Adult↗
PubMed1999

Noninvasive evaluation of Helicobacter pylori therapy: role of fasting or postprandial gastrin, pepsinogen I, pepsinogen II, or serum IgG antibodies.

OBJECTIVE: We evaluated the potential value of a change in serum IgG antibodies, fasting or meal-stimulated gastrin levels, and pepsinogen I (PGI) or pepsinogen II (PGII) levels for identifying Helicobacter pylori (H. pylori) status after antibiotic therapy. METHODS: A total of 32 men and one woman with peptic ulcer disease and documented H. pylori infection were enrolled. Fasting and 30-min postprandial blood samples were obtained at 0, 2, 7, 11, 17, 23, 27, and 39 wk of the study and were analyzed for the factors evaluated. RESULTS: Treatment was successful in 25 patients and failed in seven. Serum IgG antibodies, meal-stimulated gastrin, and both fasting and meal-stimulated pepsinogen I and II levels fell throughout the study, and pepsinogen I:II ratios increased in those whose infection was cured. The mean levels at wk 0 versus wk 7 were: fasting gastrin (fmol/ml) 12.4 and 11, meal-stimulated gastrin 26.5 and 15.4, PGI (ng/ml) 83.7 and 59, PGII (ng/ml) 24.5 and 13.6, PGI/PGII 3.5 and 4.7, and enzyme-linked immunosorbent assay value 4.8 and 4.55. The sensitivity, specificity, and positive and negative predictive values for the data analyzed using different percent changes (e.g., 80%, 50%, and 20%) were calculated. The specificity and sensitivity remained <80% at all time points. CONCLUSIONS: Despite a significant fall in serum markers of H. pylori infection in groups of individuals, no marker tested could be used to reliably determine posttherapy H. pylori status for individual patients.

Adult↗
PubMed1999

Marked differences in the frequency of microsatellite instability in gastric cancer from different countries.

OBJECTIVE: Previous studies have reported variable rates of microsatellite instability (MSI) in gastric cancer. We investigated the frequency of MSI in invasive gastric carcinoma of patients from three geographic regions. METHODS: Genomic DNA from gastric cancer and nontumor tissue from 22 Korean, 20 Colombian, and 26 U.S. patients was amplified with five microsatellite markers. RESULTS: MSI was more frequently seen in gastric cancer from Korea, affecting 50% of patients, in contrast with gastric cancers from the U.S. (7%) and Colombia (15%) (p = 0.003 and p = 0.03, respectively). MSI at one locus was significantly more frequent in gastric cancer from individuals >65 yr (p = 0.01). MSI was similarly associated with both diffuse and intestinal types of gastric cancer. CONCLUSIONS: MSI affects the two major histological types of gastric cancer, and was more frequent in gastric cancer from Korea than in the other countries, suggesting that the relative importance of different pathways of gastric carcinogenesis may vary in diverse regions of the world.

Aged↗
PubMed1999

Molecular relationships of Helicobacter pylori strains in a family with gastroduodenal disease.

OBJECTIVE: Few studies have examined the genetic relationships of Helicobacter pylori strains affecting family members. Our aim was to do so. METHODS: We characterized H. pylori isolates obtained from members of a single family presenting with various gastroduodenal diseases to examine H. pylori bacterial genetic similarity. Endoscopic evaluation with gastric mapping was performed on each individual to establish clinical and histological disease. Genomic DNA extracted from each H. pylori isolate was used to generate DNA fingerprints for each strain by REP-PCR. vacA genotypes and cagA presence were established by PCR. RESULTS: Gastrointestinal diseases among the five members of this family included gastric adenocarcinoma in a 52-yr-old man (index patient), gastric MALT-lymphoma in the 73-yr-old mother; intestinal metaplasia (IV) and atrophic gastritis in the 48-yr-old brother; intestinal metaplasia (I-III) in the 47-yr-old brother, and a duodenal ulcer scar in the 42-yr-old sister. REP-PCR DNA fingerprints of H. pylori isolates from the index patient, his mother, and both of his brothers were identical or highly similar. By contrast, the H. pylori DNA fingerprint from the sister was markedly different from the H. pylori DNA fingerprints from the other family members. All isolates had the genotype cagA-positive and vacA slb/ml mosaic genotype. CONCLUSIONS: The DNA fingerprints of H. pylori strains obtained from members of this family with malignancy or premalignant histological disease were identical or highly similar and markedly different from the H. pylori DNA fingerprint from the sibling with duodenal ulcer disease. All H. pylori isolates within the family possessed genetic markers of enhanced virulence (presence of the cagA gene and vacA sl/ml mosaicism). In addition to host genetics and environmental factors, these findings suggest that infection with genetically similar H. pylori strains is a significant factor in determining the clinical outcome of an infection with H. pylori.

Adenocarcinoma↗
PubMed1999

Sensitivity of amoxicillin-resistant Helicobacter pylori to other penicillins.

The sensitivities to penicillins and to a penicillin and beta-lactamase inhibitor combination agent were determined for Helicobacter pylori strains that were sensitive, moderately resistant, or highly resistant to amoxicillin. All strains were resistant to nafcillin and oxacillin. Moderately resistant strains showed an intermediate zone of inhibition to ticarcillin, mezlocillin, piperacillin, and amoxicillin-clavulanic acid. High-level resistance was associated with the smallest zone size for all penicillins tested.

Amoxicillin↗
PubMed1999

Proinflammatory activation of neutrophils and monocytes by Helicobacter pylori in patients with different clinical presentations.

Chronic Helicobacter pylori infection is associated with mucosal inflammation. The aim of the present study was to assess human neutrophil and monocyte activation by H. pylori strains obtained from patients with different clinical presentations. Bacterial sonicates from 12 strains were used to stimulate phagocyte upregulation of CD11b/CD18 adherence molecules assessed by fluorescence flow cytometry and oxidative burst responses assessed by chemiluminescence. A dose-dependent activation of CD11b/CD18 adherence molecules was observed with all strains on both neutrophils and monocytes. The activities were similar for strains from patients with duodenal ulceration and for strains from asymptomatic volunteers irrespective of histopathologic grades of the biopsy specimens from the antral mucosa. The neutrophil chemiluminescence response correlated with histopathologic severity. We conclude that upregulation of neutrophil and monocyte adherence molecules by H. pylori sonicates is not associated with clinical presentation of the infection.

CD18 Antigens↗
PubMed1999

DNA fingerprinting of single colonies of Helicobacter pylori from gastric cancer patients suggests infection with a single predominant strain.

In each of six gastric cancer patients, repetitive extragenic palindromic PCR DNA fingerprints of 18 single colonies of Helicobacter pylori from the gastric antrum, corpus, and cardia were identical and matched that of the parental isolate. In three additional gastric cancer patients, 17 of 18 single-colony DNA fingerprints were identical to each other and to the DNA fingerprint of the corresponding parental isolate.

Adenocarcinoma↗