Synergistic effects of glutathione and ß-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-cysteine.

Various methods have been used to remove reactive oxygen species (ROS) generated from in vitro culture (IVC) conditions that can cause cell injury or death, including the application of low oxygen (O(2)) tension and the addition of antioxidants. The beneficial effects of antioxidants and O(2) tension on IVC of porcine embryos, however, are controversial among researchers. In this study, we sought to determine the effects and optimal concentrations of antioxidants for the development of porcine embryos in an IVC system. Specifically, we examined the synergistic effects of antioxidants on development to the blastocyst stage in a culture system supplemented with L-cysteine during IVM. Of the antioxidants tested (melatonin, glutathione (GSH), ß-mercaptoethanol (ß-ME), N-acetylcysteine (NAC) and dithiothreitol (DTT)), addition of GSH (1 mM) or ß-ME (25 µM) significantly increased development to the blastocyst stage compared with the controls without antioxidant treatment (22.2 ± 4.2% for 1 mM GSH, 25.9 ± 2.2% for 25 µM ß-ME and 12-13% for the control, P<0.05). In addition, the mean cell number per blastocyst was increased by approximately 1.7-fold in the presence of GSH or ß -ME. These GSH- and ß-ME-induced increases in development to the blastocyst stage and total cell number, however, were not mimicked by melatonin, NAC or DTT, all of which are ROS scavengers. The combination of GSH or ß-ME with L-cysteine significantly reduced high O(2) tension-induced ROS production (P<0.05). These results suggest that a combination of 1 mM GSH or 25 µM ß-ME with 1 mM L-cysteine could be used for production of high quality porcine blastocysts in IVC systems.

The effect of oculo-acupuncture on recovery from ethylene glycol-induced acute renal injury in dogs.

The potential recovery effect by oculo-acupuncture (OA) on ethylene glycol-induced acute renal injury in dogs was investigated. Acute renal damage was induced by ingestion of ethylene glycol in six mongrel dogs. The dogs were assigned to control (three dogs) and experimental (three dogs) groups. The control group did not receive any treatment, while the experimental group was treated with oculo-acupuncture at kidney/urinary bladder region plus zhong jiao region of the eyes after the induction of renal damage. Serum blood urea nitrogen (BUN), creatinine, sodium (Na), chloride (Cl), and potassium (K) were measured in both control and experimental groups. The blood RBC and Hb were also examined. The serum BUN and creatinine activities in the experimental group were lower than those in the control group, the serum Na and Cl had the irregular change in both groups, and the blood Hb in the control and experimental group showed decreasing tendency. Significant differences were observed on the 3rd and 7th day in BUN, 7th day in creatinine, 2nd day in Na and Cl, and 7th day in Hb when compared to the control group. Whereas, serum K concentration and RBC in the experimental group did not change significantly. The recovery findings of the renal injury were also observed in the experimental group histopathologically. In conclusion, OA therapy (kidney/urinary bladder region plus zhong jiao region) was effective for recovery of the renal injury induced by ethylene glycol in dogs.