Interaction between Benzo[a]anthracene 7,2-dione 7-oxime (BZA) and calf thymus dsDNA using electroanalytical genosensor.

In the present study, the objective was to evaluate in situ interaction between Benzo[a]anthracene 7,2-dione 7-oxime (BZA) and calf thymus dsDNA (ct-dsDNA) using electroanalytical genosensor. Analytical techniques based on Ultraviolet/Visible (UV-Vis) spectroscopy and electroanalytical were used to investigate the interaction processes in solution and immobilized on carbon screen-printed electrodes modified with electrochemical mediator Meldola blue. In addition, was possible to evaluate the degree of damage caused to the genetic material by the analyte through of toxicity estimate (S%). The interaction evaluated by genosensor showed processes of intercalation, degradation, and breaks of the double strand of ct-dsDNA, suggesting that the interaction simulates highly toxic (values varying from 0.6 to 0.8 µA in 48 h of interaction), such as 8-oxoguanine (+0.48 V), which is a by-product of guanine oxidation. Furthermore, monitoring A (+1.10 V) after 1 h showed an S% value between 50 and 90%, indicative of high toxicity, and monitoring G (+0.85 V), which showed S>90%, indicated no toxicity after 10 min. Overall, the electroanalytical genosensor developed in a miniaturized system displayed good reproducibility and stability over time being a quick alternative for assesses the degree of toxicity between toxic xenobiotics and biologically electroactive molecules, such as DNA.

Use of two new formulations as bovine embryo manipulation solution.

This study aimed to evaluate the effect of two Embryo Manipulation Solutions (EMS and EMS supplemented) in maintenance of the viability of embryos, initially using structures derived from mice (first phase). Next, the efficiency of these solutions in routines of bovine embryo transfer was evaluated (second stage). Mice embryos were used in the stages of early blastocyst, and compact morula grades I and II. These embryos were initially randomly distributed and maintained for four hours in three solutions: Modified phosphate buffered saline (PBS; Control); EMS (treatment 1), and EMS supplemented (treatment 2). Subsequently, they were cultured in TCM 199 medium and evaluated in terms of total number of cells, morphometric characteristics, ultra structural aspects, detection of cell apoptosis, and quantification of Hsp70.3 gene expression. In the second phase, these same solutions were tested in the transfer of quality I and II bovine embryos (excellent and good). These embryos were transferred fresh to 58 recipients. The results showed that the total number of cells in embryos expanded blastocyst (ExB), the number of apoptotic cells, the cell, nuclear, nucleolar diameter and the nucleus/nucleolus ratio was similar among the treatments. The pregnancy rate shown on second phase was also similar. However, the EMS supplemented expressed more Hsp70.3 than EMS. The expression of Hsp70.3 was also greater for embryos in EMS than that of EMS supplemented. The McII embryos, EMS and EMS supplemented samples also expressed more Hsp70.3 compared to control embryos. In conclusion, the tested solutions can be used in routine embryo transfer techniques, replacing modified PBS solution as an effective media in maintaining embryo viability.