Purification and molecular analysis of BCG antigen 60

Tuberculosis remains as an important socioeconomical and medical problem throughout the world and especially in Iran. Early and timely diagnosis of pulmonary and extrapulmonary tuberculosis is vital to initiate prompt treatment. Current diagnostic methods are either slow or lack enough sensitivity or specificity. Several mycobacterial antigens are involved in the complex interaction with the immune system of the host. Their identification is important for both diagnosis and protection against mycobacteria. Antigen 60 [A60] is a thermostable antigen found in the cytosol of M. bovis and M. tuberculosis. An ELISA test using A60 is designed for diagnosis of tuberculosis with satisfactory results. In previous studies, A60 has also showed a protective effect against experimental infections and useful immunotherapeutic effects in promotion of cancer development. In the present work we tried to purify A60 from the cytoplasm of BCG. A60 was purified by exclusion gel chromatography using sepharose 4B. A60 was recognized by bidimensional immunoelectrophoresis with anti-BCG and anti-A60 antiserum, where it appears as the less mobile component. In agarose electrophoresis, A60 showed only one band but in immunodiffusion it showed two immunoprecipitinogen lines with anti-BCG anti-serum. In analyzing with dot blotting, both cytoplasm and cell wall of BCG showed positive reaction with anti-A60 anti-serum. When A60 was fractionated by SDS-PAGE and analyzed by western blot using anti-A60 antibody, 65,46,40, 38 and 35 KDa protein fractions were identified. It is concluded that A60 is a macromolecular antigen of BCG with a molecular weight of 10[6]-10[7] Da and is a lipoprotein-polysaccharide complex which contains several proteins. A60 is present in both cytoplasm and cell wall of BCG and can easily be purified from BCG vaccine using exclusion chromatography by sepharose 4B, to be used for designing diagnostic tests for TB

[Interferential currents as a treatment modality in children with nocturnal enuresis]

Background: Different treatment modalities have been recommended for enuresis which is a common childhood problem. To our knowledge, the effect of interferential currents [IF] on enuresis has not been studied previously, although there are some studies on the effect of electrotherapy in the treatment of this disorder

Materials and Methods: In this study 40 patients who were unresponsive to other modalities of treatment for enuresis, were selected for IF therapy [3 times/wk, 15 minutes per session, up to a total of 12 sessions]. Half of the children had used medications previously with no response. None of them had used an enuresis alarm

Results: The age range of the children was 5 to 15 years [mean=9.2 years] and half of them were female. Complete response was observed in 6 patients [all of them were female] and partial response [at least 50% reduction in the number of wet nights] was seen in 21 cases. Overall 67.5% of the cases had complete or partial responses. The condition remained unchanged in 23 patients who had responded for about 8 weeks after completing the treatment. There was no significant difference in clinical response between patients who had or had not received any previous medication. While, only one of the 8 patients with secondary enuresis was unresponsive, 12 out of 32 patients with primary enuresis were unresponsive to IF therapy. There was no correlation between the response and age. No complication was found during the study

Conclusion: It can be concluded that IF is effective in reducing enuresis and with more trials in the future, it may be recommended as a modality of treatment