Antioxidant effect of ergothioneine on in vitro maturation of porcine oocytes

Background@#Ergothioneine (EGT) is a natural amino acid derivative in various animal organs and is a bioactive compound recognized as a food and medicine. @*Objectives@#This study examined the effects of EGT supplementation during the in vitro maturation (IVM) period on porcine oocyte maturation and subsequent embryonic development competence after in vitro fertilization (IVF). @*Methods@#Each EGT concentration (0, 10, 50, and 100 µM) was supplemented in the maturation medium during IVM. After IVM, nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels of oocytes were investigated. In addition, the genes related to cumulus function and antioxidant pathways in oocytes or cumulus cells were investigated. Finally, this study examined whether EGT could affect embryonic development after IVF. @*Results@#After IVM, the EGT supplementation group showed significantly higher intracellular GSH levels and significantly lower intracellular ROS levels than the control group. Moreover, the expression levels of hyaluronan synthase 2 and Connexin 43 were significantly higher in the 10 µM EGT group than in the control group. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and NAD(P)H quinone dehydrogenase 1 (NQO1) were significantly higher in the oocytes of the 10 µM EGT group than in the control group. In the assessment of subsequent embryonic development after IVF, the 10 µM EGT treatment group improved the cleavage and blastocyst rate significantly than the control group. @*Conclusions@#Supplementation of EGT improved oocyte maturation and embryonic development by reducing oxidative stress in IVM oocytes.

Developmental competence of chimeric porcine embryos through the aggregation of parthenogenetic embryos and somatic cell nuclear transfer embryos

The efficiency of somatic cell nuclear transfer (NT) in pigs is low and requires enhancement. We identified the most efficient method for zona pellucida (ZP) removal and blastomere aggregation in pigs and investigated whether the aggregation of NT and parthenogenetic activation (PA) of blastomeres could reduce embryonic apoptosis and improve the quality of NT-derived embryos by investigating. Embryonic developmental competence after ZP removal using acid Tyrode's solution or protease (pronase E). The embryonic developmental potential of NT-derived blastomeres was also investigated using well-of-the-well or phytohemagglutinin-L. We analyzed apoptosis in aggregate-derived blastocysts. The aggregation rate of protease-treated embryos was lower than that of Tyrode’s solution-treated embryos (69.2% vs. 88.3%). No significant difference was observed between phytohemagglutinin-L and well-of-the-well (35.7%–38.5%). However, 2P1N showed a higher number of blastocysts compared to 3N (73.8% vs. 24.3%) and an increased blastocyst diameter compared to the control and 1P2N (274 μm vs. 230–234 μm). In blastomeres aggregated using phytohemagglutinin-L, the apoptotic cell ratio was significantly higher in 1P2N and 3N than in 3P (5.91%–6.46% vs. 2.94%, respectively). Our results indicate that aggregation of one NT embryo with two PA embryos improved the rate of blastocysts with increased blastocyst diameter.

Comparison of Safety and Efficiency of Hemodialysis Using Heparin-bound Hemophan with those of Saline Flushing Hemodialysis and Hemodialysis Using Low Dose Heparin in Patients at Risk of Bleeding / 대한신장학회잡지

OBJECTIVE: Although hemodialysis using heparin bound Hemophan (HBH-HD) has been reported to be a possible modality in patients at risk of bleeding, the efficiency and safety of HBH-HD is not certain. Therefore, we prospectively compared the safety and efficiency of HBH-HD with those of saline flushing HD (SF-HD) and HD using low dose heparin (LDH-HD) in 13 HD patients at risk of bleeding in a cross-over design. METHODS: The safety and efficiency were evaluated by measuring activated partial prothrombin time (aPTT) before and during HD, hemostasis time after needle removal, total blood compartment volume (TBCV) loss of dialyzer, urea clearance (K) and Kt/V. RESULTS: There was no difference in compression time needed to achieve hemostasis at puncture site after needle removal between HBH-HD, SF-HD and LDH-HD. During HBH-HD, there was a slight increase in aPTT at 15 min (50.6+/-4.5 sec), compared to predialysis levels (40.9+/-4.7 sec). In this cross- over study, aPTT during dialysis session was markedly higher in LDH-HD than those in HBH-HD or SF-HD (p<0.05). The loss of TBCV of the dialyzer was greater in SF-HD than HBH-HD or LDH-HD (17.4+/-1.9% vs. 12.4+/-1.4% vs. 10.1+/-1.8%). However, there was no difference in K (212.0+/-30.7 vs. 217.2+/-36.9 vs. 221.6+/- 29.5 mL/min) and Kt/V (1.22+/-0.12 vs. 1.24+/-0.16 vs. 1.26+/-0.18). CONCLUSION: We concluded that the safety and efficiency of HBH-HD are not different compared to SF-HD or LDH-HD and HBH-HD could an alternative hemodialysis method in patients at risk of bleeding.

The Influence of Low Calcium Concentration Hemodialysis on Cardiovascular Response / 대한신장학회잡지

OBJECTIVE: Arterial compliance (AC) reflects the buffering function of the vessel. Low AC caused by arterial stiffness increases pulse pressure amplitude. Therefore, Low AC must be correlated with high cardiovascular mobidity and mortality in HD patients. Dialysate calcium concentration is potentially a main determinant of serum ionized calcium level and the vasoconstriction is associated with high calcium concentration. Therefore, We conducted a study for evaluation of the interdialytic effects of treatment with a low dialysate calcium (LdCa) concentration and high dialysate calcium (HdCa) concentration on the changes of AC, BP, biochemical parameters. METHODS: Eight HD patient (mean age 45.5, sex ratio 1 : 1) were studied. The mean HD period was 3 years. Arterial Compliance, stroke Volume, SBP, DBP, PP, MAP, Ionized Ca, T-CO2, P and CaxP product were compared after treatment with a LdCa and HdCa concentration for each 10 sessions. RESULTS: AC were 0.143+/-0.076 mm2/kPa in baseline, 0.166+/-0.097 mm2/kPa in LdCa (1.25 mmol/L) dialysate, 0.142+/-0.082 mm2/kPa in HdCa (1.75 mmol/L) dialysate. SBP, DBP, MAP and PP were 157.75+/-15.97, 94.25+/-9.48, 114.12+/-10.56, 63.50+/-10.87 mmHg in baseline and 135.25+/-13.00, 78.75+/-11.24, 98.37+/-15.14, 56.50+/-5.95 mmHg in LdCa dialysate and 160.50+/-15.36, 94.05+/-10.34, 115.75+/-9.64, 62.00+/-15.71 mmHg in HdCa dialysate. Ionized Ca were 4.66+/-0.40 mg/dL in baseline, 4.45+/-0.28 mg/dL in LdCa dialysate and 4.65+/-0.43 mg/dL in HdCa dialysate. However, there were no changes of other biochemical parameters. CONCLUSION: Treatment with LdCa dialysis, by minimizing the risk for LdCa-induced hypocalcemia, may have a beneficial role in the prevention of the ongoing reduction of arterial compliance in HD patients and thus improve cardiovascular prognosis.