Coenzyme Q10 and melatonin protect cryopreserved equine sperm against lipoperoxidation.

The objective of the experiment was to evaluate the effect of the addition of different concentrations of the antioxidants Coenzyme Q10 (CoQ10) and melatonin to equine semen freezing diluent, alone or in combination, during the cryopreservation process. Twenty ejaculates (n = 5 stallions) were divided in groups: Control (C, without the addition of antioxidants), melatonin 0.75 mM (MEL1), melatonin 1.5 mM (MEL2), CoQ10 40 µg/mL (Q1), CoQ10 200 µg/mL (Q2), and CoQ10 40 µg/mL+ 0.75 mM melatonin (Q1 +MEL1). Q1 and Q2 groups demonstrated intact plasma membrane and high mitochondrial membrane potential after 30 (M-30) and 60 (M-60) min of incubation compared with the control group (Q1: 64.8 % ± 9.9 %, Q2: 65.2 % ± 10.5 %, C: 55.1 % ± 10.0 %; M-30 and Q1: 63.3 % ± 10.4 %, Q2: 64.6 % ± 10.8 %, C: 53.1 ± 10.6 %; M-60; P < 0.05). Melatonin conferred greater membrane stability at all evaluated times compared with the control group (MEL1: 42.1 % ± 6.0 %; MEL2: 44.0 % ± 6.7 %, C: 35.9 % ± 5.9 %; M-0; MEL1: 40.8 % ± 5.6 %; MEL2: 42.6 % ± 7.2 %, C: 33.1 % ± 6.6 %; M-30 and MEL1: 37.5 % ± 7.4 %; MEL2: 39.1 % ± 7.2 %; C: 31.3 % ± 6.5 %; M-60; P < 0.05). The use of antioxidants alone or in combination resulted in lower levels of lipoperoxidation at all times evaluated compared with in the control group (P < 0.05). In conclusion, CoQ10 and melatonin were effective in the cryopreservation of equine semen by decreasing lipoperoxidation and promoting a higher percentage of spermatozoa with a high mitochondrial potential, total and progressive motility, and prevention of membrane lipid disorder.

Use Low Ozone Dosages has Positive Effects on the Cooling and Cryopreservation of Equine Semen.

The aim of this study was to determine an ozone dosage capable of inducing pro-oxidation, and to verify its action on sperm cells during the process of cooling and cryopreservation of equine semen. In this study, we evaluated the ozone concentrations of 2µg/mL,15µg/mL, 30µg/mL e 60 µg/mL added in equine semen cooling and freezing extenders. Samples were evaluated for sperm kinetics patterns, function of sperm structures and lipid peroxidation. In the experiment, the concentration of 15 µg/mL showed higher total and progressive motility when comparing to control (60.3±3 and 40.7±3.4 vs. 54.9±4 e 35.0±4.4, respectively, P < .05) at M24 of cooling; The concentration of 2 µg/mL showed higher percentage of intact plasma and acrosomal membrane when comparing to control at M24 (51.1±3.6 vs. 46.1±3.9, P < .05), M24 after 30 minutes of incubation (43.4±3.1 versus 32.4±2.6, P <.05). The concentration of 2 µg/mL showed higher percentage of intact plasma and acrosomal membrane (P <.05) comparing to control at moments M0 (43.5±5.0 vs. 36.3±3.5), M30 (41.0±3,7 vs. 35.3±2,9) e M60 (39.0±7.0 vs. 31.4±5.4). Thus, it can be concluded that low doses of ozone can lead to a positive response in the sperm kinetics patterns and sperm structures after sperm storage at low temperatures. Higher concentrations (30 and 60 µg/mL) were harmful in the cooling and cryopreservation of equine semen.

Training and standardization of simulated patients for multicentre studies in clinical pharmacy education.

OBJECTIVE: To evaluate the training and standardization methods of multiple simulated patients (SPs) performing a single scenario in a multicenter study. METHODS: A prospective quasi-experimental study, using a multicenter approach, evaluated the performance of five different individuals with the same biotype during a simulation session in a high-fidelity environment. The SPs training and standardization process consisted of four steps and six web or face-to-face mediated: Step 1: simulation scenario design and pilot test. Step 2: SPs selection, recruitment and beginning training (Session 1: performance instructions and memorization request.) Session 2: check the SPs' performances and adjustments). Step 3 and session 3: training role-play and performance's evaluation. Step 4: SPs' standardization and performances' evaluation (Sessions 4 and 5: first and second rounds of SPs' standardization assessment. Session 6: Global training and standardization evaluation. SPs performance consistency was estimated using Cronbach's alpha and ICC. RESULTS: In the evaluation of training results, the Maastricht Simulated Patient Assessment dimensions of SPs performances "It seems authentic", "Can be a real patient" and "Answered questions naturally", presented "moderate or complete agreement" of all evaluators. The dimensions "Seems to retain information unnecessarily", "Remains in his/her role all the time", "Challenges/tests the student", and "Simulates physical complaints in an unrealistic way" presented "moderate or complete disagreement" in all evaluations. The SPs "Appearance fits the role" showed "moderate or complete agreement" in most evaluations. In the second round of evaluations, the SPs had better performance than the first ones. This could indicate the training process's had good influence on SPs performances. The Cronbach's alpha in the second assessment was better than the first (varied from 0.699 to 0.978). The same improvement occurred in the second round of intraclass correlation coefficient that was between 0.424 and 0.978. The SPs were satisfied with the training method and standardization process. They could perceive improvement on their role-play authenticity. CONCLUSIONS: The SPs training and standardization process revealed good SPs reliability and simulation reproducibility, demonstrating to be a feasible method for SPs standardization in multicenter studies. The Maastricht Simulated Patient Assessment was regarded as missing the assessment of the information consistency between the simulation script and the SPs provision.