RESUMO
Background: Type 4 pili [T4P] is an important virulence factor of Pseudomonas aeruginosa [P. aeruginosa]. T4P pass the outer membrane through a large oligomeric channel made of a single PilQ protein that is most highly conserved at their C-termini. To develop a functional vaccine that can be used in clinical application, the secretin domain of the PilQ [PilQ380-706] was produced as a recombinant protein
Methods: A 981 bp fragment of C-terminal of the pilQ secretin [pilQ1138-2118] from was designed into the prokaryotic expression vector pET28a. The presence of the pilQ1138-2118 gene in the recombinant construct [pET28a/pilQ] was assessed by double digestion and PCR. After transformation, expression of the recombinant PilQ was induced by addition of IPTG. The expressed recombinant protein was purified by a modified method using a HisTrap affinity column and finally confirmed by SDS-PAGE. The functional activities of the produced PilQ380-706 confirmed by Western blot analysis and twitching inhibition assay
Results: The PCR and enzymatic digestion results showed the presence of the pilQ1138-2118 gene in the construct. The protein electrophoresis showed that the molecular weight of the recombinant PilQ380-706 is approximately 37 kDa. The Western blot analysis confirmed the specificity of specific IgG against the PilQ380-706 protein. The PilQ380-706 protein showed high biological activity in all of these standard assays
Conclusion: Since, the PilQ380-706 protein plays an important role in the biogenesis of pili; and thus, the primary establishment of P. aeruginosa; it seems that it can be used as a candidate vaccine or an adjuvant in the future studies
RESUMO
The age-dependent worsening of lipid profiles increased the risk of cardiovascular disease in males. Eating fruits and vegetables is known to be beneficial on lipid profile. The objective of this research was to delineate whether drinking daily squeezed apple juice [AJ] modulates lipid profile and antioxidant enzymes in the animal model that gonadectomized. In this experimental study, 28 male rats were equally divided among the following 4 groups: group I [control], orchidectomized [ORX], ORX +AJ 10%, and ORX+ AJ 25%. After 60 days of drinking AJ, superoxide dismutase [SOD], alkaline phosphatase [ALP], cholesterol, LDL, HDL and triglycerides in serum were evaluated. Data analyzed by SPSS-16 and computed using one way ANOVA followed by Tukey multiple comparison test. P < 0.05 was statistically significant. In the ORX group compared with the sham group serum SOD and ALP activities decreased [P < 0.05], while serum cholesterol, triglycerides, LDL increased but HDL decreased in ORX groups [P < 0.05]. In contrast to the ORX group, drinking AJ decreased [P < 0.05] concentration of LDL, TG and cholesterol in the serum. AJ also increased [P < 0.05] serum HDL, SOD and ALP activities compared with the ORX group