1. Helicobacter pylori arginase inhibits nitric oxide production by eukaryotic cells: A strategy for bacterial survival Alain P. Gobert, David J. McGee, Mahmood Akhtar, George L. Mendz, Jamie C. Newton, Yulan Cheng, Harry L. T. Mobley, and Keith T. Wilson Presented by Mihaela C. Badea-Mic

2. So, Who Am I? Gram negative bacteria inside the stomach and duodenum The Helicobacter Foundation

3. Who discovered me? 1983-Campylobacter pyloridis 1997- Tomb et al sequenced the HP genome The Helicobacter Foundation, 2005

4. Epidemiology Prevalence (USA) African-American Hispanic Eastern Europeans UBIQUITOUS 50% world population The Helicobacter Foundation, 2005

5. Pathophysiology The most common route of H. Pylori infection Oral to oral Fecal to oral PS: watch your pets!

6. Histology The Helicobacter Foundation, 2005

7. Diagnosis ・ Breath test - Based on the detection of the products of urea The Helicobacter Foundation, 2005

8. Diagnosis- cont. ・ Esophagogastroduodenoscopy with biopsy ・ H. Pylori fecal antigen ・ H. Pylori serology

9. Diseases The Helicobacter Foundation, 2005

10. Diseases- cont. Gastric and duodenal ulcers Gastric cancer ( 90%) Non-ulcer dyspepsia Weird syndromes

11. Treatment Antidiarrheals – bistmuth Antibiotics - metronidazole, tetracycline, amoxicilin Proton pump inhibitors - omeprazole, lansoprazole H2 receptor blockers – ranitidine, famotidine

12. My survival strategies D. S. Merrel et al, 2004

13. My virulence factors D. M. Monack et al, 2004

14. Details D. M. Monack et al, 2004

15. And now…finally the paper Activated macrophages produce NO using L-arginine as a substrate H. Pylori arginase competes with NOS2 for the substrate H. Pylori converts the substrate to urea and L-ornithine, not NO rocF gene encodes arginase Mutations in rocF gene helps the NO to kill H. Pylori

16. Experiment # 1A Hypothesis - the activated macrophage production of NO is inhibited by wt H. Pylori at physiologic L-arginine concentrations

17. Experiment #1A – cont. Conclusion: -only the wt H. Pylori inhibited NO released - the arginase deficient H. Pylori did not inhibit

18. Experiment # 1B Conclusion: adding more substrate will stop the competitive inhibition between H. pylori and activated macrophages

19. Experiment # 2 Hypothesis – H. Pylori wt compete for the same substrate with activated macrophages. This substrate is L-arginine.

20. Experiment # 2 –cont. Conclusion -the wt H. Pylori uses L-arginine decreasing the macrophage NO production because of the loss of the substrate - the rocF mutant does not use the L-arginine

21. Experiment # 3 Hypothesis – the macrophage production of NO is regulated by the H. Pylori arginase independently of iNOS expression Conclusion- both H. Pylori induce iNOS mRNA expression

22. Experiment # 4 Hypothesis – The bacterial arginase inhibits the release of NO by preformed iNOS in macrophages.

23. Experiment # 4-cont Conclusion: the wt H. Pylori inhibits macrophage NO production by the preactivated cells

24. Experiment # 5 Hypothesis – Only viable H.pylori can inhibit the release of macrophage NO Conclusion : only live H. Pylori can consume the substrate in order to inhibit the iNOS

25. Experiment # 6 Hypothesis – Inhibition of the host cell NO production is a survival strategy for H. Pylori. Conclusion – the wt H. Pylori has an increased rate of survival in comparison with rocF mutant H. Pylori in the presence of macrophages.

26. Experiment # 6 A

27. Experiment # 6B Conclusion: The rocF mutant H. Pylori survives in the presence of iNOS -/- macrophages.

28. Summary Bacterial arginase evolutionary adaptation survival strategy for H. pylori gastric mucosa protection

29. Don’t let your kids kiss us! The Helicobacter Foundation, 2005

30. Invitation

31. That is all I had to say… The End!