Assessment of helicobacter pylori viability by flow cytometry

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. 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. 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. 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

[Soluble CD26 and CD30 concentration in cutaneous leishmaniasis]

Introduction: Cutaneous leishmaniasis [CL] is the most common form of leishmaniasis that usually heals spontaneously with unsightly scar but rarely non-healing lesion of CL develops which is refractory to all types of therapy. It is shown that Th1 and Th2 response is associated with healing and non-healing form of the diseases, respectively. On the other hand, it is reported that CD26 and CD30 are associated with Th1 and Th2 types of response, respectively. In some diseases, there is a relationship found between level of CD26 and CD30 and Th1 and Th2 responses

Objective: The goal of this study was to determine the concentration of soluble CD26 and soluble CD30 [sCD26 and sCD30] as a possible marker for Th1/Th2 response in CL

Materials and Methods: The blood samples were taken from 36 patients with healing form of the lesion and 10 patients with non-healing form of CL who were referred to Razi hospital or Center for Research and Training in Skin Diseases and Leprosy in Tehran during 2003-2004. As a control blood samples were taken from 23 volunteers with no history of CL. In this study, the concentration of sCD26 and sCD30 were measured in plasma by ELISA method

Results: The results showed that the plasma levels of sCD26 and sCD30 were significantly higher in non-healing form of the disease than healing form of CL or control group [p<0.05]. The level of sCD30 was more prominent than sCD26. There was no significant difference in the level of sCD26 or sCD30 markers in healing form of CL compared to normal control group

Conclusion: Overall it seems that the level of sCD30 might be a useful marker to study the immune response in CL, which needs to be studied further

Comparison of validity of microlymphocytotoxicity and flow cytometry methods with PCR for HLA-B27 antigen typing

The Human Major Histocompatibility Complex [MHC] plays a crucial role in transplantation, transfusion, paternity test and assessment of susceptibility to some diseases associated with HLA-B27. Three of the most fashionable methods for determination of HLA antigens in clinical and research laboratories are microlymphocytotoxicity [MLCT], flowcytometry and polymerase chain reaction [PCR].The purpose of this study was to compare the sensitivity, specificity, and positive and negative predictive values of MLCT and flowcytometry methods with PCR as a gold standard method in determination of HLA-B27 antigen. In the present study, all three above-mentioned techniques have been used for 36 patients suffering from ankylosing spondylitis and 31 healthy volunteers. Specific antisera and monoclonal antibodies against HLA-B27, and allele specific PCR have been used in MLCT, flowcytometry and PCR methods respectively. The results show that sensitivity, specificity, positive and negative predictive values of MLCT method as compared with PCR technique were 83.3%, 100%, 100% and 88.1% respectively. Moreover, sensitivity, specificity, positive and negative predictive values of flowcytometry compared to the PCR technique were 100%, 94.6%, 93.8% and 100% respectively. Based on the results, the flowcytometry method in determination of HLA-B27 is more valid than MLCT in this regard, particularly in research programs. The similarity between the results of our study and those studies done in Europe suggests the probability of resemblance between HLA-B27 subtypes in Europe and in Iran

Carpal tunnel syndrome among long-term hemodialysis patients

Among 123 patients on chronic hemodialysis, 13 patients with carpal tunnel syndrome [11 bilateral] were detectd. They tended to be 12 years older and had been on hemodialysis two years longer than patients without C.T.S. Duration on hemodialysis appeared to have a causative effect for the development of C.T.S. among females, while impaired calcium-phosphorous metabolism appeared to be implicated among males. The presence of shoulder pain and cystic radiolucencies correlated with C.T.S. Of six hands which were explored surgically, amyloid deposition was found in three. The site of vascular access, sex, underlying renal disease, efficacy of dialysis and predialytic beta 2-microglobulin level had no relation to the development of C.T.S