Assessment of Helicobacter pylori Viability by Flow Cytometry

Background: Flow cytometry is a rapid, sensitive, and reliable method for determination of bacterial viability. Here we assayed the capability of flow cytometry to detect Helicobacter pylori viable cells in both forms of spiral and coccoid. Methods: Viable bacteria stained with Rhodamin 123 and fluoresced with laser beam of 488nm. The rate of Rh123 absorption was determined in both forms of bacteria. Results: In positive control that consisted of live bacteria, the rate of rh123 absorption was at highest, but negative control that consisted of dead bacteria, the rate of Rh 123 absorption was at lowest absorption. This method showed that non-culturable coccoid forms of H. pylori, which could resist environmental stresses, were alive and might be responsible for bacterial transmission and failure in disease treatment. Conclusion: Due to simplicity, reliability, and sensitivity of flow cytometry, this method is preferred to other expensive and no reliable methods such as autoradiography, PCR and Electron microscopy used for assessment viability

Th1/Th2 Cytokines in Psoriasis

Background: The aim of this study was to determine the Th1 and Th2 serum cytokines, in patients with psoriasis and to com¬pare their cytokine levels with those of normal control subjects. Methods: Serum levels of Interferon-gamma (IFN-γ), Interleukin-2 (IL-2), Interleukin-4 (IL-4), and Interleukin-10 (IL-10) were measured by enzyme linked immunosorbent assay in 40 patients with psoriasis and in 40 normal controls. Results: Compared with control subjects, patients with psoriasis had elevated levels of IFN-γ and IL-2 (P<0.001). In addi¬tion a positive correlation was found between the levels of IFN-γ, IL-2 and disease severity. Conclusion: Th1 secreting inflammatory cytokines may contribute to the pathogenesis of psoriasis

Comparison of Apoptosis, Circulating Levels of Tumor Necrosis Factor-α and Tumor Necrosis Factor Type-I Receptor in Iranian Patients with Sepsis and Normal Controls

Background and Aim: The present study was designed to compare the cell death, circulating levels of tumor necrosis factor-α and tumor necrosis factor type-I receptor in Iranian patients with sepsis and normal controls.Materials and methods: Twenty-two patients with sepsis were included in this study. After blood collection, the serum circulating levels of TNF- and TNFRI measured with ELISA kits. The PBMCs isolated from blood samples and proportion of apoptotic cells measured by flowcytometry at the time of blood draws (0 time) and after 24-h incubation. PBMCs incubated at 37°C in culture (spontaneous apoptosis) and in the presence of rTNF that is capable of inducing apoptosis in activated T cells expressing the TNF family of receptors.Results: PBMCs obtained from the patients showed significantly higher (P<0.001) proportion of apoptotic cells than PBMCs of controls at 0 time, indicated that a higher fraction of PBMCs were undergoing apoptosis in vivo in patients but not in controls. After 24-h incubation, spontaneous ex vivo apoptosis of PBMCs was nearly as high as that of TNF- induced apoptosis, indicating that activated T cells had been preprogrammed in vivo to die.Discussion and Conclusion: The circulating levels of both TNF- and TNFRI showed significantly higher in patients (P<0.001) than controls and this increase is proportional (r=0.908) in both indicating that TNFRI may have a protective effect in the early stage of sepsis