p53 and mitogen-activated protein kinase pathway protein profiles in fresh and frozen spermatozoa.

Sperm or testicular tissue cryopreservation is performed in cases of male infertility as a treatment for the preservation of fertility. When these sperm cells are used in assisted reproductive techniques, fertilisation rates, developmental and implantation potential of embryos decrease and the abortion rates increase. In the present work, differences of both phosphorylation and expression levels of p53 and Mitogen-activated protein kinases (MAPK) proteins were analysed in 61 individual sperm samples before and after cryopreservation. We observed that p53 protein residue at Ser 15 was phosphorylated after cryopreservation. Because MAPK pathway activations may be involved in p53 phosphorylation, MAPK/ERK, Stress-activated protein kinases (SAPK)/JNK and p38MAPK proteins were also investigated. Analysis showed that p38MAPK phosphorylations increased significantly. However, ERK and JNK expressions and phosphorylations decreased, although the differences were not statistically significant. According to our results, it may be suggested that cryopreservation process activates p53 via p38 MAPK pathway that subsequently causes apoptosis, which may be related to sperm parameters.

Effect of the new legislation and single-embryo transfer policy in Turkey on assisted reproduction outcomes: preliminary results.

New legislation concerning assisted reproduction treatments was introduced in Turkey in March 2010 in order to reduce the number of multiple pregnancies. This new legislation limits the number of embryos to be transferred to one under 35 years of age in the first or second treatment cycles and to two in the third or further cycles or for 35 and older ages. The aim of this multicentre study was to investigate the effect of this new law on clinical pregnancy and multiple pregnancy rates. Outcomes were compared in equal periods of 2.5 months before and after the new law, and further investigation was conducted for two different age groups: <35 and ≥ 35. The clinical pregnancy rates decreased from 39.9 to 34.5% and multiple pregnancy rates decreased from 23.1 to 5.3% (P<0.001) for the overall population. The outcomes of the <35 age group and ≥ 35 age group were also similar to that of the overall population. These results suggest that under the new legislation multiple pregnancy rates are significantly reduced without causing a significant decline in the pregnancy rates.

Cycle outcome can be predicted by the growth profile of untransferred cleavage-stage embryos.

Excess embryos obtained from intracytoplasmic sperm injection cycles may be cultured and observed until day 5 if the couple receiving treatment do not want them to be cryopreserved. In order to investigate the correlation between blastocyst formation in extended culture and pregnancy outcome, 194 patients treated in two separate IVF units were examined retrospectively. The patients were separated into two groups: group 1 with at least one blastocyst formed in culture, and group 2 with no blastocyst formation. The pregnancy rates were 60.0% and 41.7% for groups 1 and 2, respectively. The pregnancy rate in group 1 was statistically significantly higher than in group 2 (P = 0.01). The results suggest that the developmental potential of embryos obtained from a single assisted reproduction treatment cycle may be similar and that blastocyst formation in vitro may help to predict the pregnancy outcome of that cycle.

Predictors of intracytoplasmic sperm injection (ICSI) outcome in couples with and without male factor infertility.

AIM: To find out the predictors of ICSI outcome. METHODS: Forty-three pregnancies in 100 consecutive ICSI cycles. RESULTS: Every 1,000 pg/ml increase in hCG-day E2 (OR = 0.46, CI: 0.25-0.83, p = 0.01) and 1% decrease in the rate of normal sperm morphology (OR = 0.81, CI: 0.67-0.98, p = 0.03) caused a significant decrease in clinical pregnancy rate and live birth rate (respectively, OR = 0.5, CI: 0.32-0.96, p = 0.03, OR = 0.66, CI: 0.5-0.86, p = 0.002) while every increase in the number of good quality embryos transferred caused a two-time increase in the clinical pregnancy rate (OR = 2.1, CI: 1.2-4, p = 0.01). On the other hand, every increase in the number of four-cell cleavage embryos (OR = 1.02, CI: 1.002-1.04, p = 0.03) and hCG-day endometrial thickness (OR = 1.6, CI: 1.15-2.24, p = 0.005) were found to increase the live birth rate. Implantation rate (m = 8.3 +/- 14.6) was significantly lower in cases with leucocytospermia (n = 33) compared to cases without leucocytospermia (n = 67, m = 17.4 +/- 24.6, p = 0.02). CONCLUSION: Leucocytospermia, hCG-day E2 level and endometrial thickness, normal sperm morphology, and number of good quality embryos are predictors of implantation, clinical pregnancy and live birth rate following ICSI.

Bleeding patterns in women using intramuscular progesterone for luteal support in in-vitro fertilisation cycles.

This paper aims to evaluate the effects of i.m. progesterone on bleeding patterns after in-vitro fertilisation embryo transfer (IVF-ET). It is a retrospective cohort study conducted in the reproductive endocrinology and IVF department of a teaching hospital. One hundred consecutive women were studied who had undergone IVF-ET using 'long protocol' stimulation with leuprolide acetate-recombinant follicle stimulating hormone (rFSH) and who did not become pregnant. Intramuscular (i.m.) progesterone (50mg once daily) was started the day before oocyte retrieval and continued for a minimum of 12-14 days following embryo transfer. The main outcome measures were time interval between oocyte retrieval and onset of bleeding, luteal phase serum progesterone and oestradiol (E2) levels, and midluteal endometrial thickness. Of the 100 patients whose charts were reviewed, 67 bled (group A) before progesterone treatment was discontinued (17 days after oocyte retrieval) and 33 (group B) bled after progesterone treatment was discontinued (> 17 days). Mean onset of bleeding was 16.2+/-2.6 days after oocyte retrieval. Serum progesterone concentrations were similar in the two groups on the day of hCG administration, whereas progesterone concentrations (in-group B) were higher on days 7 and 15 after oocyte retrieval. No statistically significant differences were found between two groups with respect to mean midluteal endometrial thickness and mean serum E2 concentrations on days 0, 7 and 15. The results suggest that i.m progesterone administration for luteal support in assisted reproduction cycles elongates luteal phase in some patients due to supraphysiological serum progesterone levels. However, most patients start to bleed in the absence of pregnancy despite continued progesterone treatment.

A retrospective review of perioperative complications in 360 patients who had Burch colposuspension.

This retrospective study reviews intraoperative and early complications of Burch colposuspension of 360 patients. Ten patients had massive haemorrhage and 8 of them had a blood transfusion. Three patients had a haematoma. Bladder injuries were noticed in 10 patients, 3 of whom were diagnosed postoperatively. One patient had unilateral ureteral kinking. Urinary retention occurred in 20 patients for more than 10 days and 2 required catheterization for 26 and 32 days respectively. Eighteen patients had a wound infection and 4 had a wound abscess. Twenty nine patients had a urinary infection. Urinary tract injury, haemorrhage and blood transfusion were significantly more common in women having secondary surgery than those having primary surgery. Deep venous thrombosis was diagnosed in 3 patients who had a Burch colposuspension with concomitant abdominal hysterectomy. Knowledge of possible risks and complications of Burch colposuspension may help plan a better preoperative work-up of patients and may minimize the intraoperative complications and increase surgical success and patient satisfaction.