Taciana F. Soares

Evaluation of [13C]urea breath test and Helicobacter pylori stool antigen test for diagnosis of H. pylori infection in children from a developing country.

ABSTRACT The [13C]urea breath test (13C-UBT) and Helicobacter pylori stool antigen test (HpSA) for the diagnosis of H. pylori infection in children were validated. The sensitivity, specificity, and positive and negative predictive values were 93.8, 99.1, 97.8, and 98.0%, respectively, for the 13C-UBT and 96.9, 100, 100, and 98.0%, respectively, for HpSA. Both tests are appropriate for diagnosing H. pylori infection in children.

Factors Associated with Helicobacter pylori Infection by a cagA-Positive Strain in Children

Although infection with a cagA-positive Helicobacter pylori strain is considered a risk factor for the development of duodenal peptic ulcer in adults, this association has not been demonstrated in children. The presence of cagA was investigated by polymerase chain reaction in H. pylori strains isolated from 27 children with duodenal ulcer and 53 without duodenal ulcer. All patients (100%) with duodenal ulcer and 33 (62.3%) without ulcer were colonized by a cagA-positive strain (P=.00007). A cagA-positive status was also associated with a more marked macroscopic gastritis, with a greater inflammatory infiltrate of both mononuclear and polymorphonuclear cells in the antral and oxyntic gastric mucosae and degenerative and regenerative changes of the gastric mucosa. Increased cagA positivity was also associated with increased age, but no association between cagA-positive status and sex was observed.

Variation Rhythms of Lymphocyte Subsets during Healthy Aging

Immunological alterations associated with aging (immunosenescence) do not represent a simple unidirectional decline in all functions but develop as a complex remodeling of the immune system, involving multiple reorganization and developmentally regulated changes. In general, most data available about aging were obtained at particular age intervals and most of them come from Caucasian individuals from either Europe or the United States. Here, we report the frequencies of major lymphocyte subsets in healthy Brazilian individuals from 2 distinct geographic regions (Southeast and South) at several age intervals spanning a lifetime period (0–86 years). Overall, we demonstrated that changes in the frequencies of cells related to both innate and adaptive immunity clearly occur with aging in these individuals. These changes were not progressive and equally steady for all cell populations tested but instead showed an oscillatory or rhythmic behavior that was distinctive of each population at different age intervals. We also observed that abrupt changes in the frequencies of immune cells may occur in healthy individuals over 75 years old, suggesting there is an impaired flexibility of the immune system at late stages of life to sustain homeostasis via immune mechanisms. We presented reference ranges for healthy Brazilian individuals at all ages. The knowledge of these parameters in further detail will allow interventions to optimize immune function in advanced age and to improve the quality of life in the elderly.

Factors Associated with Helicobactev pylori Infection by a cagA-Positive Strain in Children

Although infection with a cagA-positive Helicobacter pylori strain is considered a risk factor for the development of duodenal peptic ulcer in adults, this association has not been demonstrated in children. The presence of cagA was investigated by polymerase chain reaction in H. pylori strains isolated from 27 children with duodenal ulcer and 53 without duodenal ulcer. All patients (100%) with duodenal ulcer and 33 (62.3%) without ulcer were colonized by a cagA-positive strain (P=.00007). A cagA-positive status was also associated with a more marked macroscopic gastritis, with a greater inflammatory infiltrate of both mononuclear and polymorphonuclear cells in the antral and oxyntic gastric mucosae and degenerative and regenerative changes of the gastric mucosa. Increased cagA positivity was also associated with increased age, but no association between cagA-positive status and sex was observed.

dupA polymorphisms and risk of Helicobacter pylori-associated diseases

The dupA of Helicobacter pylori has been suggested as a virulence marker associated with the development of duodenal ulcer disease. However, the studies performed in different geographical areas have shown that there are variations in the prevalence of dupA and its association with H. pylori clinical outcomes. Our group did not observe associations between the presence of dupA and H. pylori clinical outcomes in Brazil. On the other hand, we observed 2 mutations in the sequence of dupA that lead to stop codons: a deletion of an adenine at position 1311 and an insertion of an adenine at position 1426 of the gene. Our aim was to evaluate associations of the presence of dupA with duodenal ulcer and gastric cancer, considering dupA-positive only those H. pylori strains that do not have the mutations in the gene sequence. We also evaluated the effect of infection with a strain carrying an intact dupA on the gastric mucosa histology and IL-8 gastric levels. Colonization with strains that had the intact dupA was negatively associated with gastric carcinoma (p=0.001, OR=0.32, 95% CI=0.16-0.66). The presence of dupA was also associated with an increased degree of antral mucosa inflammation (p=0.01) and with decreased corpus atrophy (p<0.01) as well as with increased gastric mucosa IL-8 levels (p=0.04). In conclusion, the infection with a H. pylori strain containing the dupA without the stop codon polymorphisms is associated with a lower risk of development of gastric carcinoma in Brazilian subjects.

The interrelationship between Helicobacter pylori vacuolating cytotoxin and gastric carcinoma.

Objective:We aimed to determine whether cytotoxin-positive Helicobacter pylori strains are associated with gastric carcinoma.Methods:We studied 130 patients: 57 H. pylori-positive patients with gastric carcinoma, 53 H. pylori-positive patients without gastric carcinoma, and 20 H. pylori-negative subjects. The ability of H. pylori strains to produce vacuolating cytotoxin was tested in INT-407 and HeLa cells. The presence of antibodies to cytotoxin was investigated in blood serum from all subjects by immunoblotting. Fragments of the gastric mucosa from patients without gastric carcinoma and H. pylori-negative subjects were obtained for histopathological study.Results:Considering the results as a whole, 40 (70.2%) patients with and 22 (41.5%) without gastric carcinoma were colonized by cytotoxin-positive strains. Antibodies against cytotoxin were not observed in the serum from 17 (29.8%) gastric carcinoma patients and from 31 (58.5%) patients without gastric carcinoma. H. pylori strains isolated from these patients did not produce cytotoxin in vitro. In regard to cytotoxin positivity, a significant difference was observed between patients with and without gastric carcinoma (p= 0.004; odds ratio [OR]: 3.3; 95% confidence interval [CI]: 1.4–7.9). Higher scores of mononuclear (p= 0.0001) and polymorphonuclear (p= 0.000003) cells were observed in the antral mucosa from H. pylori-positive patients without gastric carcinoma infected by cytotoxin-positive strains than in those harboring cytotoxin-negative strains.Conclusion:Cytotoxin-producing H. pylori strains were more frequently observed in patients with gastric carcinoma and this aspect emphasizes the role of cytotoxin in the genesis of the tumor.

The role of IFN-gamma and IL-4 in gastric mucosa inflammation associated with Helicobacter heilmannii type 1 infection

Although Helicobacter heilmannii infection is less common than H. pylori infection in humans, it is considered to be of medical importance because of its association with gastritis, gastric ulcer, carcinoma, and mucosa-associated lymphoid tissue lymphoma of the stomach. However, there have been no studies evaluating the role of the Th cell response in H. heilmannii gastric infection. We evaluated the participation of pro-inflammatory and anti-inflammatory cytokines, IFN-gamma and IL-4, in H. heilmannii gastric infection in genetically IFN-gamma- or IL-4-deficient mice. The serum IFN-gamma and IL-4 concentrations were determined by ELISA. The gastric polymorphonuclear infiltrate was higher (P = 0.007) in H. heilmannii-positive than in H. heilmannii-negative wild-type (WT) C57BL/6 mice, whereas no significant inflammation was demonstrable in the stomach of H. heilmannii-positive IFN-gamma(-/-) C57BL/6 mice. The degree of gastric inflammatory cells, especially in oxyntic mucosa, was also higher (P = 0.007) in infected IL-4(-/-) than in WT BALB/c mice. Serum IFN-gamma levels were significantly higher in IL-4(-/-) than in WT BALB/c mice, independently of H. heilmannii-positive or -negative status. Although no difference in serum IFN-gamma levels was seen between H. heilmannii-positive (11.3 +/- 3.07 pg/mL, mean +/- SD) and -negative (11.07 +/- 3.5 pg/mL) WT BALB/c mice, in the group of IL-4(-/-) animals, the serum concentration of IFN-gamma was significantly higher in the infected ones (38.16 +/- 10.5 pg/mL, P = 0.04). In contrast, serum IL-4 levels were significantly decreased in H. heilmannii-positive (N = 10) WT BALB/c animals compared to the negative (N = 10) animals. In conclusion, H. heilmannii infection induces a predominantly Th1 immune response, with IFN-gamma playing a central role in gastric inflammation.

Prospects for the development of scientific research in Medical Education

This text intends to present potential themes of scientific research, in the area of medical education, to professionals involved in teaching and medical training. The need to promote teaching competencies in this area is pressing in order to understand it as a fruitful field of research, and execute experimental studies.

S1644 Helicobacter pylori Infection Displays a Mixed Inflammatory/Regulatory Cytokine Profile in Children Characterized by Increased Gastric TH1, TH2, TH17 and Treg Cytokines

H. pylori CagA protein with higher number of EPIYA-C segments or EPIYA-D motif has been associated with precancerous lesions and gastric cancer, which needs to be confirmed in different regions. Recently, it has also been shown that CagA contains polymorphic CM located downstream the EPIYA-C or D segment that mediates interaction of CagA with PAR1b/MARK2 and inhibits PAR1b kinase activity disrupting tight junctions and epithelial cell polarity. Because CagA+ status is associated with increased IL-1β, IL-8 and IL-12 release, studies attempting to demonstrate In Vitro associations between IL-8 levels and the number of EPIYA-C segments have been done, but the results are discordant. Here, we evaluated whether the number of EPIYA-C segments is associated with the risk of H. pylori-related diseases adjusting for gender and age. We also evaluate the type and number of CM sequences in respect to the risk of diseases and gastric cytokine levels. We studied 521 patients with gastritis (G, n=250, 114 cagA-), duodenal ulcer (DU, n=149, 19 cagA-) and gastric carcinoma (GC, n=125, all cagA+). The number of EPIYA-C segments and CM was determined by PCR followed by sequencing. The gastric levels of IL-1β, IL-8 and IL-12 were evaluated by ELISA (Biosource, Camarillo). Patients with GC (41%) were more often infected with strains with more than one EPIYA-C segment than the patients with G (18.5%) and DU (17.0%). In the logistic analysis, the number of EPIYA C segments (OR=3.05, 95%CI=1.64-5.66, p .23), DU (p>.29) and GC (p>.20). Neither the number nor the Eastern CM pattern was associated with the diseases or cytokine levels, but both associated with diffuse (90.1%, p=.01), but not intestinal (70.1%) tumor type. Infection with strains with more than one EPIYA-C segment was significantly associated with corpus intestinal metaplasia (p<.000) and atrophy (p=.02). In conclusion the number of EPIYA-C segments but not CM is associated with increased risk of GC. Although CagA enhances IL-8 and IL-12p70 gastric levels, the number of EPIYA-C segments and CM does not affect the cytokine production. Of note, increased Eastern CM that is associated with junctional cell defects was associated with diffuse type tumor. CNPq/FAPEMIG/INCT.

Factors Associated withHelicobacter pyloriInfection by acagA‐Positive Strain in Children

Although infection with a cagA-positive Helicobacter pylori strain is considered a risk factor for the development of duodenal peptic ulcer in adults, this association has not been demonstrated in children. The presence of cagA was investigated by polymerase chain reaction in H. pylori strains isolated from 27 children with duodenal ulcer and 53 without duodenal ulcer. All patients (100%) with duodenal ulcer and 33 (62.3%) without ulcer were colonized by a cagA-positive strain (P=.00007). A cagA-positive status was also associated with a more marked macroscopic gastritis, with a greater inflammatory infiltrate of both mononuclear and polymorphonuclear cells in the antral and oxyntic gastric mucosae and degenerative and regenerative changes of the gastric mucosa. Increased cagA positivity was also associated with increased age, but no association between cagA-positive status and sex was observed.