Pregnancy outcome according to male diagnosis after ICSI with non-ejaculated sperm compared with ejaculated sperm controls.

The aim of this study was to describe pregnancy outcome in couples who had undergone ICSI using non-ejaculated sperm from men with non-obstructive azoospermia, obstructive azoospermia and aspermia compared with the outcome of ICSI with ejaculated sperm from men with severe oligozoospermia, treated during the same time period. This nationwide cohort study included all children born after ICSI with non-ejaculated sperm in Norway, from when the method was first permitted in Norway in April 2004 to the end of 2010, resulting in 420 pregnancies and a total of 359 children. In 235 of these children, the father was diagnosed with obstructive azoospermia, in 72 with non-obstructive azoospermia, in 31 with aspermia, and in 21 the male cause was unclassifiable. The control group consisted of 760 children from 939 pregnancies conceived by ICSI with ejaculated sperm. Sex ratio, birth weight, rate of pregnancy loss and congenital malformations were not significantly associated with sperm origin or the cause of male factor infertility.

Blastocyst transfer and gender: IVF versus ICSI.

PURPOSE: To investigate the difference in sex ratio in our centre after day 5 embryo transfer among neonates delivered after fresh in vitro fertilization (IVF), fresh intra cytoplasmatic sperminjection (ICSI) and frozen embryo replacement (FER) compared to expected sex ratio in normal population. METHODS: Retrospective data after IVF/ICSI/FER therapy from 1995 - 2007 and data from the literature on similar subjects. RESULTS: Among a total of 420 births after day 5 transfer, there were 225 male and 195 female babies. However, after ICSI alone 30 were female and 21 male. Adding all available data from the literature on gender ratio after long term culture, the ratio of male births to total births (0.545) was significantly (p = 0.004) different from the expected ratio (0.515). CONCLUSIONS: It appears that significantly more males are born than expected after day 5 transfer in IVF. However, a subgroup analysis in our data comparing IVF, ICSI and FER revealed more females born after ICSI and FER, suggesting that further research is needed in this area.

PP072. Development of polyphenols-containing nanoparticles for the treatment of vaginal inflammation.

INTRODUCTION: Vaginal inflammation during pregnancy is linked to miscarriage, preterm labour, placental dysfunction and perinatal complications. Despite the availability of several anti-inflammatory drugs, there is no general consensus regarding safe formulation especially for use in early pregnancy. On the other hand, polyphenols especially "curcumin" from Turmeric (powdered rhizome of Curcuma longa) and "catechins" from green tea have been traditionally used and widely discussed (>16,000 citations for curcumin and >24,000 for catechin in SciFinder) as pharmacologically active anti-inflammatory components and considered as safe. However, due to their poor solubility, low bioavailability and stability, their clinical application remains to be limited. OBJECTIVES: Our aim was to develop a non-teratogenic and non-toxic polyphenol-bearing nanoparticles which can be safely used in early pregnancy for the treatment of vaginal inflammation. METHODS: Vesicle-based liposomal formulations for polyphenols (curcumins and catechins) were developed and standardized. Liposomal curcumins and catechins were characterized in respect to their stability, size and entrapment efficiency. Antioxidant activities were evaluated by DPPH/ABTS+/O2.- and SOD activities. Anti-inflammatory activities were evaluated by measuring LPS-induced NO and pro-inflammatory cytokines such as IL-1B and TNF-A in macrophages (J774.1) and LPG-induced cytokines IL-8 in human vaginal cells lines (End1/E6E7, Ect1/E6E7, VK2/E6E7). Antioxidant and anti-inflammatory activities of curcumin/catechin and their corresponding vesicle-based formulation were compared. RESULTS: Phospholipid-based nanoparticles (approx. 200 nm) were found to be stable and incorporating high amount of curcumins or catechins. Liposomal curcumin was found to be 2- to 6-folds more potent than curcumin in inhibiting NO production and pro-inflammatory cytokines in macrophages. IL-8 was inhibited up to 67% by the liposomal curcumin in human vaginal cell lines (End1/E6E7, Ect1/E6E7, VK2/E6E7) as compared to curcumin. However, similar vesicle formulations of catechins were not significantly superior to non-vesicle formulations of catechins. CONCLUSION: Standardized liposomal curcuminoids have potential as an effective and safe topical formulation for the treatment of vaginal inflammation in early pregnancy.

Differential pH in embryo culture.

OBJECTIVE: To determine the optimum pH in sequential media for embryo culture around the fertilization-zygote stage and cleavage stage, with use of a mouse embryo assay. DESIGN: Experimental laboratory study. SETTING: University Hospital and University Research Unit. ANIMAL(S): F1 hybrids between CD1 female and BDF male mice. INTERVENTION(S): Fertilized, one-cell mouse embryos were cultured 5 days in test media where pH was changed at defined time intervals. MAIN OUTCOME MEASURE(S): Percentage of good-quality embryos, defined by strict morphology. RESULT(S): A significantly improved development was observed when pH was as high as 7.30 before the pronuclear stage and lowered to pH 7.15 during the cleavage period. CONCLUSION(S): Good embryo development is consistent with two different pH values in sequential culture media. This could have important implications for embryo culture in human IVF.

New media for culture to blastocyst.

OBJECTIVE: To develop sequential media for extended culture of embryos in human IVF, with use of a mouse embryo assay. The BlastAssist system was developed. Blastocyst culture has been our routine method since 1994, using earlier media IVF and M3 in sequence. DESIGN: Experimental laboratory study. SETTING: University Hospital and university research unit. ANIMAL(S): F1 hybrids between CD1 female and BDF male mice. INTERVENTION(S): Two-cell mouse embryos were cultured 4 days in test media; 1 day in phase 1 medium followed by 3 days in phase 2 medium. MAIN OUTCOME MEASURE(S): Percentage of expanded and hatched blastocysts. RESULT(S): Compared with a two-cell block in the old culture system, more than 80% went to the blastocyst stage in the new media. The new medium system was based on a heavily modified low-glucose Earle's balanced salt solution. Additions were pyruvate, lactate, amino acids, human serum albumin, and Synthetic Serum Replacement. In contrast to regular tissue culture cells, embryos required an increased concentration of EDTA. The final media produced a blastocyst frequency of 94% and a day 4 hatching frequency of 71%. CONCLUSION(S): With the high blastocyst frequency demonstrated here, the combination of blastocyst culture and single ET should be an effective means of treatment in patients and could help to eliminate multiple gestations. The BlastAssist system is commercially available.

Noninvasive cerclage for the management of cervical incompetence: a prospective study.

OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of a noninvasive cerclage pessary in the management of cervical incompetence. METHODS: This is a prospective cohort study of all pregnant women treated for cervical incompetence during a 4-year period. Women with known risk factors for preterm delivery had transvaginal ultrasonography every 2-3 weeks after 17-19 weeks of gestation. Those with progressive shortening of cervix diagnosed before 30 weeks were treated with a cerclage pessary when the cervical length was < or = 25 mm. The pessary was electively removed at 34-36 weeks. The course and outcome of pregnancy were recorded. RESULTS: Thirty-two women were treated with a cerclage pessary. There were nine twin and two triplet pregnancies. Fifteen (47%) had two or more risk factors for preterm delivery. The mean gestational age at cerclage was 23 (17-29) weeks, cervical length 17 (5-25) mm. Two women required delivery before the onset of labor due to severe intrauterine growth restriction and one due to HELLP syndrome. These were excluded from further analysis. In the remaining 29 women, the interval between cerclage and delivery was 10.4 (2-19) weeks, mean gestational age at delivery 34 (22-42) weeks, and birth weight 2,255 (410-4,045) g. Thirteen (45%) women delivered before 34 weeks. There were a total of 35 live-born infants and four intrapartum fetal deaths (all between 22 and 25 weeks gestation). All women complained of increased vaginal discharge, but no other significant complications were observed that could be attributed to the use of pessary. CONCLUSION: Cerclage pessary may be useful in the management of cervical incompetence. Whether it can be a noninvasive alternative to surgical cerclage merits further investigation.