Molecular detection of host cytokine expression in Helicobacter pylori infected patients via semi-quantitative RT-PCR.

BACKGROUND: Helicobacter pylori (Hp) is a bacterium recognised as a main causative agent for the development of chronic active gastritis, peptic ulcer disease, gastric adenocarcinoma and primary gastric lymphoma. OBJECTIVE: Determination of the levels of IFN-gamma (pro-inflammatory) and IL-4 (anti inflammatory) cytokine expression as indicators of Th1 and Th2 immune responses in gastric cancer (GC) and non gastric cancer (Non GC) dyspeptic patients by gene specific RT-PCR. MATERIALS AND METHODS: Biopsy specimens were collected from three groups of gastric cancer (GC=18), non ulcer dyspepsia (NUD = 38) and peptic ulcer patients (PUD=20). Total RNA was extracted and complementary DNA was synthesised. PCR amplification was performed for HPRT, IFN-gamma and IL-4 cytokines and the intensity of each band was measured by densitometry and normalized against HPRT expression as a house keeping gene. RESULTS: Comparison of the results from different groups of patients indicated that IFN-gamma gene expression was similar in nonGC dyspeptic patients (NUD and PUD groups; 3.38 +/- 0.57,3.43 +/- 0.41, respectively) whereas, in GC patients, it was significantly higher than others (5.52 +/- 0.59; P < 0.0001). On the other hand, IL-4 gene expression showed no significant difference between NUD and GC patients (2.81 +/- 0.43,2.3 +/- 0.12 respectively), whereas the expression rate of this cytokine was significantly higher in PUD patients (3.7 +/- 0.1; P 0.05). Our data indicate an association between Th1 and Th2 immune responses and the development of gastric cancer and peptic ulcer disease respectively.

Detection of atypical cultivable canine gastric Helicobacter strain and its biochemical and morphological characters in naturally infected dogs.

Helicobacter-like organisms are frequently found in canine stomachs, but the relationship between such organisms and gastric pathology has not been established. However, some such organisms have zoonotic importance. The aims of this study were to evaluate the morphological and biochemical characteristics of cultivable canine gastric Helicobacter-like organisms (GHLOs) in pets and stray dogs and their prevalence in these two groups of dogs. Specimens were taken by gastroscopy from 30 clinically healthy stray dogs and 30 pet dogs. Cultures were positive from biopsies of 11/30 of stray and 6/30 of pet dogs. The isolated Helicobacters were observed by light microscopy and studied by biochemical, physiological and PCR analysis. Some of the isolated GHLO's displayed atypical shapes that were similar to Helicobacter pylori or Helicobacter acinonychis in stray dogs' cytological examinations. They had 2-3 helices and were smaller than other canine GHLOs. One of these atypical Helicobacter strains was cultured. It was not possible to distinguish such strains by routine PCR and biochemical evaluations. Electron microscopy showed a smaller Helicobacter (2 microm in length) with 2 or 3 helixes. This study demonstrates that not all canine gastric Helicobacters are 5-15 microm in length, as has been previously proposed, and portrays the need for further investigation of canine GHLOs.

Prevalence of Helicobacter pylori vacuolating cytotoxin and its allelic mosaicism as a predictive marker for Iranian dyspeptic patients.

Helicobacter pylori infects the majority of the population in the developing countries. However, the rate of gastrointestinal complications such as peptic ulcers and gastric malignancies has no parallel with the infection. In order to determine whether cytotoxin (vacA) and its allelic polymorphism can serve as screening markers for such a population, H. pylori strains were isolated from one hundred and thirty two dyspeptic patients. H. pylori genomic DNA was extracted and underwent PCR-amplification for the cytotoxin alleles. Genotyping of the signal sequence region of the vacA gene identified 68% (70 out of 103) of patients with non ulcer dyspepsia (NUD) and 79% (23 out of 29) of the patients with peptic ulcer disease (PUD) possessing the s1 genotype. S1 strains were significantly more prevalent among patients with PUD as compared to the NUD (p < 0.05). In regard to the middle region, 55% of the patient isolates belonged to the m2 genotype with no correlation to disease. The s1m2 genotype was the most prevalent among all patients and significantly correlated with the PUD group (p < 0.05).