Cytotoxic Effects of Verbascoside on MCF-7 and MDA-MB-231

Objectives: Verbascoside, also known as acteoside/kusaginin, has attracted a great attention due to its pharmacological features. In this study, we aimed to determine the cytotoxic effects of pure verbascoside isolated from Phlomis nissolii L. plant in both MCF-7 and MDA-MB-231 cell lines in vitro. Materials and Methods: MCF-7 and MDA-MB 231 cells were treated with verbascoside (100, 48, 25, 10, 1, 0.5, and 0.1 µM) for 24, 48, and 72 hours. Cytotoxic effect of verbascoside in MCF-7 and MDA-MB-231 cells was assessed using TEBU-BIO cell counting kit 8. Results and Conclusion: IC50 values for 24, 48, and 72 h verbascoside exposure of MCF-7 cells were determined as 0.127, 0.2174, and 0.2828 µM, respectively. R2 values were calculated as 0.9630, 0.8789 and 0.8752, respectively. Two-Way ANOVA multiple comparison test results showed that 100 µM verbascoside has the highest cytotoxic effect on MCF-7 breast cancer (BC) cells after 72 h of exposure. IC50 values for 24, 48 and 72 h verbascoside exposure of MDA-MB 231 cells were determined as 0.1597, 0.2584 and 0.2563 µM, respectively and R2 values were calculated as 0.8438, 0.5107 and 0.9203, respectively. Two-Way ANOVA multiple comparisons test results showed that 100 µM verbascoside has the highest cytotoxic effect on MDA-MB 231 BC cells after 24, 48 and 72 h of exposure.

Evaluation of subgroups of the human sperm hypoosmotic swelling test in normozoospermic male cases with recurrent fertilization failure: a prospective case-controlled study.

PURPOSE: To compare subgroups of the human sperm hypoosmotic swelling test subgroups in both recurrent fertilization negative infertile cases with normal semen analysis and fertilization positive controls. METHODS: This was a prospective case-controlled study performed with normospermic 33 previously fertile male (secondary infertility) and 41 infertile men who had undergone two or three unsuccessful in vitro fertilization attempts. HOS test was investigated in 4 subgroups including HOS 1, HOS 2, HOS 3, and HOS 4 according to the degree of sperm tail swelling and compared between the two groups. RESULTS: Four subgroups were compared and statistical significance was demonstrated in HOS 1, HOS 3 and HOS 4 tests (p < 0.001) in fertile and infertile men. Highest HOS 1 and lowest HOS 4 grades were determined in Group A. However, no statistical significance was determined between two groups in HOS 2 test which was minimal swelling in sperm tails. CONCLUSIONS: HOS 1, HOS 3 and HOS 4 subgroups of HOS test are reliable and useful methods providing important information regarding the sperm function. A high HOS test 1 grade plus a low HOS test 4 grade should suggest a fertility problem, despite a normal semen analysis. HOS test subgroups provide additional information in normospermic cases with unexplained infertility.

Serum anti-Müllerian hormone level as a predictor of poor ovarian response in in vitro fertilization patients.

PURPOSE: To evaluate the clinical value of day 3 serum anti-Müllerian hormone (AMH) compared with day 3 serum follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG) day estradiol (E2) levels and antral follicle count (AFC) in the prediction of poor ovarian response in controlled ovarian hyperstimulation (COH). METHODS: AMH, FSH and AFC on day 3 as well as hCG day E2 levels were determined in 164 subjects. Receiver operating curve analyses and area under curves (AUC) of the study parameters were performed. Predictive values of the levels of day 3 AMH, FSH, AFC, and hCG day E2 as clinical parameters of ovarian response to COH were studied. RESULTS: Thirty-eight women were defined as poor responders. The day 3 AMH and hCG day E2 levels and AFC of normal responders were significantly higher than those of the poor responders. In predicting poor response, the AUC of day 3 AMH level was significantly higher than that of day 3 FSH level but was similar to the hCG day E2 level. Day 3 AMH, FSH and hCG day E2 levels and AFC were found to predict a poor response. Day 3 AMH and hCG day E2 levels were more predictive compared with day 3 FSH level and AFC. The cut-off level of AMH was ≤2 with a sensitivity of 78.9% and a specificity of 73.8%. CONCLUSION: Day 3 AMH has the ability to predict a poor response to COH and it is more predictive than day 3 FSH and AFC.

Does granulosa cell apoptosis have a role in the etiology of unexplained infertility?

OBJECTIVE(S): To compare granulosa cell apoptosis in patients with unexplained infertility and tubal factor. Accelerated granulosa cell apoptosis may be the cause of unexplained infertility. STUDY DESIGN: Setting was IVF-ET Unit of Cerrahpasa Medical Faculty, Istanbul University. GnRH analogs and gonadotropins were used for ovulation induction in patients with unexplained infertility (n=15) and tubal factor (n=15) undergoing in vitro fertilization and embryo transfer (IVF-ET) procedures. Following HCG injection and follicular aspiration, apoptosis of granulosa cells was assessed using the in situ DNA nick end labelling method and apoptosis rate was further determined by flow cytometry. Apoptosis rates were compared between two groups. Mann-Whitney's U-test and Student's t-test were used for statistics. RESULTS: Apoptosis rate was significantly higher in the unexplained infertility group (33.20 +/- 35.62% versus 10.10 +/- 17.23%). CONCLUSION(S): Granulosa cell apoptosis seems to have a role in the etiology of unexplained infertility.