Elicitation of IgY in chicken egg yolk by recombinant fragments of UreC and its efficacy against Helicobacter pylori

Helicobacter pylori multiplies and causes infection in human gastric mucosal layer. New approaches have focused on using specific treatments, such as immunotherapy, to limit this infection. Urease, as one of the most important virulent and antigenic factors of the bacterium, is a suitable target for this purpose. In order to prepare recombinant proteins, the synthetic genes for total ureC protein [UreCt] and its N [ureCn] and C [ureCc] terminal fragments were ligated into pET28a. The recombinant proteins were expressed in E. coli BL21[DE3]. White leghorn hens were injected with the purified recombinant proteins. IgY recovered from egg yolk, using PEG precipitation. Finally, urease neutralizing ability of the antibodies was evaluated by urease activity assay in presence of the purified IgY. SDS-PAGE analysis revealed a good expression and purification of the recombinant proteins. Indirect ELISA observation demonstrated high antibody titer in sera and egg yolks and high ability of IgY Anti-UreCt and IgY Anti-UreCc antibodies in recognition of urease subunit C. Anti-UreCT and Anti-UreCc IgYs were more potential H. pylori urease inhibitors than Anti-UreCn. While all three UreC fragments induce prophylactic responses. UreCt and UreCc possess almost equal responses. Anti-UreCc IgY has advantage of smaller size and is preferred for its activity and easier protein recovery and purification process. These features emphasize on importance of simpler, easier and cost effective antibody production

Familial relations and recurrence pattern in nephrolithiasis. New words about old subjects

While medical and surgical approaches to urolithiasis are different for single and recurrent stone former [RSF], the RSF definition itself is commonly overlooked. Moreover, despite consensus on association between family history [FH] and urolithiasis, more epidemiologic evidence is required to clarify the nature of this relationship. Our purpose was to propose a more precise definition of RSF, and also to investigate how family history may affect urolithiasis. Using a multistage stratified sampling in 4 seasonal phases, 6127 subjects with imaging-proven urolithiasis were detected in 12 Iranian regions. The FH of urolithiasis and the average interval between episodes [cycles] were determined by an informed interview. Of 6127 patients with the mean age of 41.8 +/- 15.1 years, 42% had FH, and 22.2% were RSF of whom 61% were men. The patients with FH had a greater chance of recurrence [OR = 1.2, 95% Confidence Interval [CI], 1.1 to 1.4]. Furthermore, patients with positive FH had more episodes [P = .0001], comparable cycles and younger ages at the onset [P = .02] than those patients without a FH. In the RSF group, the 90[th] percentiles of the cycle were 60 months and the estimated mean stone cycle for the population was 25.34 months [99% CI, 23.0 to 27.7]. Family history seems very common in Iranian population and is a risk factor for recurrence. Moreover, RSF could be identified by the estimated average cycle in the population [25.3 months] or by the percentiles

Production of IgY in chicken egg yollk against helicobacter pylori urease using UreC recombinant protein

Helicobacter pylori is a spiral, microaerophilic gram negative bacterium, that multiplies and causes infection in human gastric mucosal layer. H.pylori infection, followed by destruction of gastric epithelial tissue, leads to gastric chronic inflammation, which can cause gastric and peptic ulcers. New approaches have focused on using specific treatments, such as immunotherapy, to eradicate this infection. Urease, as one of the most important virulent and antigenic factors of the bacterium, is a suitable target for this purpose. This study is aimed at production of specific IgY against urease UreC subunit. In this study, initially for preparing recombinant UreC, after purification of the genomic DNA, ureC gene was amplified by polymerase chain reaction [PCR]. The PCR product was ligated to pET28a. The recombinant protein was expressed followed by transformation of recombinant construct into E.coli BL21DE3. SDS-PAGE analysis was carried out and the recombinant protein was purified by Ni-NTA affinity chromatography. The purified recombinant protein was injected to hens. IgY recovered from egg yolk, was purified by PEG precipitation at >70% purity. The purified IgY was analyzed by ELISA and SDS-PAGE. SDS-PAGE analysis revealed a good expression and >70% purification of the recombinant protein. ELISA observation demonstrated high immunogenicity of the recombinant protein. With a view to higher potential of IgY-HpUc in recognition of UreC subunit, the results are in favour of the oral administration of the IgY obtained from hens immunized by H.pylori may provide a novel approach to the management of H.pylori infections