Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus).

The pacu is one of the most important species for Brazilian fish farming and is considered emerging in the global aquaculture. Despite its importance, no effective tool for evaluation of the adaptive immune response of this species has been developed. Therefore, this study aimed the development and standardization of indirect ELISA for the measurement of pacu antigen-specific antibodies using polyclonal rabbit anti-pacu IgM used as detector antibody. For this purpose was isolated and purificated of pacu IgM using mannose-binding protein affinity chromatography and produced specific polyclonal antibodies against heavy and light chains pacu IgM, that showed a molecular weight of 72 kDa and 26 kDa, respectively. Polyclonal antibodies obtained demonstrated specificity with heavy and light Ig chains of pacu serum in western blotting. These polyclonal antibodies allowed the development of an indirect ELISA assay of high sensitivity and specificity for the detection and quantification of pacu IgM antibodies immunized with bovine IgG. In conclusion, this approach has great potential to improve the monitoring of vaccine-induced immune responses and help develop immunodiagnostic and epidemiological studies of infectious diseases in pacu systems.

Effects of electroacupuncture on experimental periodontitis in rats.

BACKGROUND: Acupuncture has shown the capability of modulating the immuno-inflammatory response of the host. This study aims to evaluate the effects of electroacupuncture (EA) on ligature-induced periodontitis in rats. METHODS: Thirty-two animals were divided into four groups: 1) control; 2) experimental periodontitis (EP); 3) sham-treated (EP/EA-sham); and 4) treated with EA (EP/EA). For the EP groups, a ligature was placed around the right mandibular first molars at day 1. Sessions of EA or EA-sham were assigned every other day. For EA treatment, large intestine meridian points LI4 and LI11 and stomach meridian points ST36 and ST44 were used. EA-sham was performed in off-meridian points. Animals were euthanized at day 11. Histomorphometric and microtomographic analyses were performed. Immunolabeling patterns for the receptor activator of nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase (TRAP) were assessed. Expressions of interleukin (IL)-1ß, matrix metalloproteinase (MMP)-8, IL-6, and cyclooxygenase (COX)-2 messenger RNAs (mRNAs) were evaluated by quantitative reverse transcription-polymerase chain reaction. Data were analyzed statistically (P <0.05, analysis of variance). RESULTS: Histomorphometric and microtomographic analyses demonstrated that group EP/EA presented reduced alveolar bone loss when compared to group EP (P <0.05). Reduced RANKL immunolabeling and fewer TRAP-positive multinucleated cells were observed in the EA-treated group in relation to group EP. No differences were observed in OPG expression among groups. EA treatment decreased the genic expression of IL-1ß and MMP-8 (P <0.05), increased the mRNA expression of IL-6 (P <0.05), and did not modify the genic expression of COX-2 in animals with EP (P >0.05). CONCLUSION: It can be concluded that EA reduced periodontal tissue breakdown and the expression of some proinflammatory mediators and a proresorptive factor in EP in rats.

Short-term evaluation of grafts fixed with either N-butyl-2-cyanocrylate or screws.

PURPOSE: N-butyl-2-cyanoacrylate (NB-Cn) is an alternative method for onlay graft fixation and might be efficient for preserving the graft volume. Our aim was to analyze the gene expression and mineralized tissue variations of calvarial bone grafting fixed in the mandible with either NB-Cn or a titanium screw (TiS). MATERIALS AND METHODS: New Zealand rabbits had bilateral calvarial grafts fixed at both sides of the mandible with either NB-Cn or a TiS. The rabbits were sacrificed at 4 and 8 days, and micro-computed tomography analysis was performed. For molecular analysis, the gene expression of interleukin-6, interleukin-10, and tumor necrosis factor-α was assessed. Quantification using real-time polymerase chain reaction was performed. Statistical analysis was performed using the paired Student t test (P < .05). RESULTS: Bone graft fixation with NB-Cn promoted superior volume and density preservation. The percentage of mineralized tissue at the center portion and border of the graft was very similar (NB-Cn, 50.6% ± 8.3% and 50.3% ± 10.6%, respectively) and superior than in the TiS group (32.5% ± 3.5% and 33.8% ± 6%, respectively). Genes from the NB-Cn group were upregulated compared with those in the TiS group at the initial phases of bone healing (4 days), with the profile reversed at the 8-day point. At day 8, the osteoclastogenesis-related genes were upregulated in the TiS group. CONCLUSIONS: Onlay bone grafts fixed with screws induced more inflammation during the initial remodeling process than did NB-Cn. The differences in the incorporation into the host bed suggest that the use of adhesives for graft fixation will promote superior volume and density preservation.