Anti-CASP3 monoclonal antibody Knockout Validated

Lupeol is a triterpene with various pharmacological properties. This study investigated the effect of lupeol on the in vitro development of bovine embryos. Oocytes (270 per group, 1620 in total) obtained from slaughterhouse-derived ovaries were matured and fertilized in vitro and then cultured for 8 days in a humidified atmosphere of 5% CO2 in air at 38.5 degrees C. The in vitro maturation medium was supplemented with 0.5, 1.0, 2.0, 3.0, and 4.0 mu M lupeol. Treatment with 2.0 mu M lupeol significantly (P < 0.05) improved blastocyst development. Hoechst 33342 staining and terminal deoxynucleotidyl transferase dUTP nick end labeling showed that treatment with 2.0 mu M lupeol improved blastocyst quality by increasing the total cell number and reducing the apoptotic cell number. Confocal microscopy confirmed that treatment with 2.0 mu M lupeol significantly (P < 0.05) reduced the level of 8-oxoguanine, an indicator of reactive oxygen species. Lupeol treatment also significantly attenuated protein expression of nuclear factor-kappa B subunit 1 (NEKB1), cyclooxygenase (COX) 2, and CASP3. Real-time PCR analysis of nitric oxide synthase 2, NFKB1, COX2, CASP3, and BCL2-associated X protein supported the immunofluorescence data. In conclusion, lupeol is a potent antioxidant that improves bovine embryo development in vitro. (C) 2017 Elsevier Inc. All rights reserved.

This study compared slow freezing and vitrification of ovarian tissue by evaluation of histological changes, WNT signaling pathway and apoptotic genes expression. Ovarian tissue was obtained from women aging 27-38 years old. Ovarian cortex from each patient was divided into three pieces and randomly grouped as slow freezing, vitrification and control groups for investigation of WNT signaling gene expression and beta-CATENIN presence as well as histological studies. The stromal structure of all ovaries were preserved. The number of secondary follicles decreased in vitrified group (P < 0.05). WNT-3, beta-CATENIN, FZD-2 and GSIC-3 beta expressions were significantly higher in slow frozen and vitrified groups, compared to control group (P < 0.05). On the contrary, AXIN1 expression in slow frozen samples were significantly lower than that of the vitrified and control group. The expression of apoptotic genes, excluding CASP3, was significantly decreased in slow-frozen samples (P < 0.05). Conversely, BAX:BCL-2 percentage significantly increased in vitrification versus slow freezing and control(P < 0.05). Follicles in slow frozen samples displayed nuclear and cytoplasmic beta-CATENIN staining, while control and vitrification groups only showed beta-CATENIN protein in the cytoplasm. The presented data show that slow freezing results in a better preservation regardless of the type of follicle. Therefore, it is concluded that slow freezing is still an ideal method for ovary cryopreservation. (C) 2017 Elsevier Inc. All rights reserved.