Perurethral transvesical route for oocytes retrieval: an old technique for a new indication in oncofertility.

We aimed to evaluate the safety and efficiency of the peruretheral transvesical oocyte retrieval in oncofertility. We conducted a retrospective comparative study in our assisted reproductive technologies center. STUDY GROUP: 28 pubertal young women affected by malignancies, referred for fertility preservation and refusing transvaginal (TV) procedure. CONTROL GROUP: 28 infertile patients, aged less than 25 years, who have undergone in vitro fertilization with TV oocyte retrieval. The ovarian stimulation was significantly longer on the study group. There was no difference between the two groups regarding mean number of collected metaphase II oocytes. One patient of the study group had a transient dysuria. These preliminary data suggest that, in oncofertility, peruretheral transvesical oocyte retrieval is an alternative when the TV route is refused or not feasible.

Basal follicle stimulating hormone level correlated to age is a good prognostic criterion for the outcome of intracytoplasmic sperm microinjection.

BACKGROUND: Age and increased FSH serum level in women are prognosis criteriae associated with decreased fertility. OBJECTIVE: The aim of this study was to investigate whether age-specific FSH concentration can be a predictor of the outcome of ovarian stimulation in women undergoing IVF. METHODS: A total of 676 women undergoing their first IVF cycle over a 3-year period were included in this retrospective cohort study. Patients were grouped according to age (< or ≥  38 years), and within each age range, patients were grouped into bFSH quartiles (< or ≥  9.6 mUI/L). We have considered four study groups: group A (Age < 38 years and FSH < 9.6 m UI/l), group B (Age < 38 years and FSH ≥ 9.6 m UI/l), group C (Age ≥ 38 years and FSH < 9.6 m UI/l), group D (Age ≥ 38 years and FSH ≥ 9.6 m UI/l). The outcome measures in each group included: consumed quantity of gonadotrophin, poor response, cycle cancellation, oocyte yield, number of embryos obtained, embryonic quality (grade 1 embryo), as well as, fertilization, implantation, clinical pregnancy and childbirth rates. Analysis of the Results compares laboratory parameters and ICSI Results, based on a statistical analysis that is essentially descriptive. RESULTS: High bFSH levels in young patients (< 38 years) predicts a higher poor response (p < 0.0001), higher stopped cycles (p < 0.0001), lower oocyte yield (p < 0.0001) and lower embryos obtained (p < 0.0001) in IVF cycles but does not translate to either lower pregnancy, childbirth or implantation rates. In old women high FSH level does not influence ICSI outcome but may increase poor response (p <0.01) and stopped cycles (p < 0.0001). In each age group, the rate of spontaneous miscarriage does not increase according to FSH level. The pregnancy rate and child birth rate are better in young women with high FSH levels than in older women with normal FSH levels (p < 0.05). CONCLUSION: The findings of this study suggest that basal FSH concentrations when correlated to age is a good predictive factor of ovarian response for assisted reproductive treatment. In young women a high FSH level may affect laboratary parametres but not pregnancy rate. In old women normal FSH level does not improve ICSI outcome but may increase "avorted" cycles. Pregnancy rate and child birth rate are better in young women with high FSH levels than in older women with normal FSH levels.

Progesterone for Luteal Phase Support in In Vitro Fertilization: Comparison of Vaginal and Rectal Pessaries to Vaginal Capsules: A Randomized Controlled Study.

BACKGROUND: In IVF, Luteal phase support is usually performed using vaginal progesterone. A part of patients using this route reports being uncomfortable with this route. We tried to study whether the rectal route could be an effective alternative and associated with less discomfort. PATIENTS AND METHODS: A prospective randomized controlled study. All patient were eligible for IVF treatment for infertility. After oocyte pickup, 186 patients were allocated to one the following protocols for luteal phase support: (i) rectal pessaries group: natural progesterone pessaries administered rectally 200 mg three times a day, (ii) vaginal pessaries group: natural progesterone pessaries administered vaginally 200 mg three times a day), and (iii) vaginal capsules group: natural micronized progesterone capsules administered vaginally 200 mg three times a day. On the day of pregnancy test, patients were asked to fill in a questionnaire conducted by an investigator in order to assess the tolerability and side effects of the LPS treatment taken. The primary endpoint was the occurrence of perineal irritation. RESULTS: Fifty eight patients were assigned to the rectal pessaries group, 68 patients to the vaginal pessaries group, and 60 patients to the vaginal capsules group. All patients adhered to their allocated treatment. Implantation and clinical pregnancy rates per transfer did not differ between the three groups. Perineal irritation, which was our primary endpoint, was the same for all the three groups (respectively 1.7 % versus 5.9 % versus 11.7%). Regarding the other side effects, more patients experienced constipation and flatulence with the rectal route, whereas more patients reported vaginal discharge in the vaginal capsules group. CONCLUSION: Rectal administration for luteal phase support is effective and well accepted alternative to vaginal route.

Vascular endothelial growth factor (-460 C/T, +405 G/C, and +936 C/T) polymorphisms and endometriosis risk in Tunisian population.

The vascular endothelial growth factor (VEGF), a major angiogenic factor, is known to play an important role in the development of endometriosis. The aim of this study was to investigate the association of three VEGF (-460 C/T, +405 G/C, and +936 C/T) polymorphisms with the risk of endometriosis in the Tunisian population. This study includes 105 women with endometriosis and 150 women with no laparoscopic evidence of disease. Genotyping of the VEGF -460 C/T, +405 G/C, and +936 C/T polymorphisms were performed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The distribution of genotypes (P = 0.006) and allele (P = 0.0009) frequencies of the +936 C/T polymorphism was significantly different between patients and controls. Patients with stages III-IV endometriosis showed a higher VEGF + 936T allele frequency than controls (P = 0.0001). However, the distribution of genotypes and allele frequencies of the VEGF -460 C/T and +405 G/C polymorphisms did not differ significantly between endometriosis patients and controls. These findings suggest that the VEGF +936 C/T polymorphism may be a risk factor for endometriosis development and the VEGF +936 T allele is associated with an increased risk of stages III-IV endometriosis in the Tunisian population.

Glutathione S-transferase M1 and T1 gene polymorphisms and risk of endometriosis in Tunisian population.

OBJECTIVE: An association between endometriosis and the glutathione S-transferase M1 (GSTM1)- and GSTT1-related genes has been proposed on account of the detoxification properties of the GST enzymes. The aim of the present study was to investigate whether the polymorphisms and null mutations are associated with the susceptibility to endometriosis. METHODS: The study included 105 women with endometriosis and 150 healthy women with no laparoscopic evidence of disease. Genotyping of the GSTM1 and GSTT1 gene polymorphisms was performed by Multiplex-PCR. RESULTS: There was a significant association of GSTM1 null and GSTT1 null genotypes with endometriosis both when studied alone (P = 0.001 and P = 0.03, respectively) and in combination (P = 0.00002). CONCLUSION: The findings suggest that the GSTM1 and GSTT1 gene deficiency predisposes to endometriosis in a Tunisian population and can confer a significant increased risk when the GST null genotypes are combined.

Failure of in vitro fertilization: prognosis criteriae.

BACKGROUND: age, obesity and increased FSH serum level in women are prognosis criteriae associated with decreased fertility and adverse Assisted Reproductive Technologies (ART) outcomes. OBJECTIVE: To assess the effect of age, FSH and BMI on pregnancy rate in ICSI. METHODS: A retrospective and comparative study of 500 women who underwent ICSI cycle during the study period from January 2004 to December 2005. Age, FSH and BMI were compared in two groups of patients: Those achieving a pregnancy: The "pregnancy+" group and those failing to have a pregnancy: The "pregnancy-"group. For each of previous parameters ROC curve and logistic regression study were performed. RESULTS: age was significantly lower in "pregnancy+" group (32,4±3,9 years vs 33,7±4,8 ans ; p=0,005). Analysis of ROC curve and logistic regression study show that for age, the most discriminative cut-off for predicting pregnancy is 38 years (Se=7,5%, Sp=75,6%) (AUC=0,572; p=0,02) (OR=2,1 ; LR+=6,7 ;IC[1 ;1,4] ; p0,009). FSH was significantly lower in "pregnancy+" group (5,5±1,8UI /L vs 6,2±3UI/L, p=0,003). Analysis of ROC curve and logistic regression study show that for FSH, the most discriminative cut-off for predicting pregnancy is 9UI/L (Se=3,7%, Sp=85.2%) (AUC=0,539 ; p=0,03) (OR=3,6; IC[1,4 ;9,3]; LR+ =10,1; p=0,003). BMI was also significantly lower in "pregnancy+" group (24,7± 3,6 kgm-2 vs 27,1±4,5 kgm-2 ; p<10-3). The most discriminative cut-off for predicting pregnancy is 25,4 kgm- 2 (Se=31,7%, Sp=33,3%) (AUC=0,663 ; p<10-3) (OR=4; IC[2,1 ;7,7]; LR+ =19,38; p<10-3). CONCLUSION: age, FSH and BMI affect markedly the prognosis of ICSI. We found significantly lower Pregnancy rates in older women (> 38 years), in women with elevated FSH (> 9UI/L) or elevated BMI (> 25,4 kgm-2). Our results can be used when counseling and before including patients in an IVF program, to give them probability of success and weight loss required to optimize chances of pregnancy.

Impact of gamete cell quality on the outcome of intracytoplasmic sperm injection.

BACKGROUND: the quality of the gametes used for an intracytoplasmic microinjection of spermatozoïde is a significant factor which can influence pregnancy rates. AIM: To assess the effect of conventional sperm parameters, origin of spermatozoa and oocyte quality on pregnancy rate in ICSI. METHODS: A retrospective and comparative study of 500 women who underwent ICSI cycle during the study period from January 2004 to December 2005. Conventional sperm parameters (count, motility and morphology) and oocyte quality (mature and immatures oocytes) was compared in two groups of patients: Those achieving a pregnancy: The "pregnancy+" group and those failing to have a pregnancy: The "pregnancy-" group. RESULTS: Among the conventional sperm parameters, only spermatozoa count after preparation was significantly higher in "pregnancy+" group (p=0,02). We found significantly more pregnancies in ejaculated and epididymal sperm groups than in the testicular one (p<10-3). The number of oocyte retrieved was significantly higher in "pregnancy+" group (13,9±7 vs 10,6± 7,6 ; p<10-3) with mainly mature oocyte (metaphaseII) (9,1±5,5 vs 6,6±5,4 ; p<10-3). Analysis of ROC curve and logistic regression study show that for mature oocyte, the most discriminative cut-off for predicting pregnancy is 4 (OR=2,1 ; LR+=6,7 ;IC[1 ;1,4] ; p0,009). CONCLUSION: Conventional parameters of ejaculated sperm have almost no influence on pregnancy rates in ICSI. Testicular sperm seem to have worse results. The number of oocytes retrieved and the proportion of mature oocytes (metaphaseII) affect markedly the prognosis.

Seminal vesicle agenesis: An uncommon cause of azoospermia.

Seminal vesicle malformations are a rare cause of obstructive azoospermia, often associated with other internal genitalia and upper urinary tract birth defects. We report 5 new cases of seminal vesicle agenesis in men presenting with hypospermia and azoospermia. Imaging showed seminal vesicle unilateral agenesis in all patients. The remaining seminal vesicle was hypoplastic in 3 cases, dilated in 1 case and with abnormally thick content in another case. Vas deferens agenesis was observed unilaterally in 2 patients and bilaterally in 2 other patients. No renal malformations were detected. Genetic study showed in all cases a 46 XY karyotype without any microdeletions. A single heterozygous cystic fibrosis transmembrane regulator gene mutation was diagnosed in 1 man, but not found in his partner. Intracytoplasmic sperm injection using sperm from a testicular biopsy was performed in 3 couples, without success.

[Short vs long agonist protocols in poor responders undergoing IVF]. / Résultats de I'ICSI chex les mauvaises répondeuses : protocole court vs protocole long.

AIM: Compare among poor responders: stimulation results, laboratory parameters and the final IVF results by assessing 2 different stimulation protocols: the long agonist protocol and the short agonist protocol. METHODS: An analytical retrospective study carried out over of period of 2 years: January 2006 and December 2007. During this period, a total of 1192 IVF cycles of ICSI type were performed in 892 patients. INCLUSION CRITERIA: short agonist or antagonist stimulated patients protocols and presenting two of the three following criteria: 1- Patients aged more than 38 years with an FSH plasmatic rate on the 3rd day of the cycle 9.5 UI/ml. 2- Antral follicle count (AFC) 5 for both ovaries. 3- Failure of anterior ovary stimulation: abandonment of cycle or 3 oocytes at data collection in a previous cycle. EXCLUSION CRITERIA: PCOS or single ovary. RESULTS: 65 patients, undergoing 92 attempts of ICSI cycles have been included in this study. Long agonist protocol was performed in 48 cases and Short agonist protocol was performed in 44 cycles. Both groups were comparable as to age (40,09 ± 6, 59 vs 41, 04 ± 1,71 years; NS), BMI (25,2±3,92 vs 25,35±4,09 Kgm-2 ; NS), infertility type (primary 41% vs 59%;NS ; ou secondary 58% vs 40,9% ; NS), FSH (9,98±2,42 vs 10,01±2,75 ; NS) and antral follicle count on day 3 (4,13±1,12 vs 3,8±1,16 FA ; NS). The estradiol rate, dosed on the onset day was significantly higher in the short protocol group (1534,27±1034,34 vs 1133,31±1053,58 pg/ml; p=0.034). However, the consumed quantity of gonadotrophins was lower in the short protocol group (1550±235,45 vs 1725,55±450,35 UI, p=0.01). A total of 13 cycles was stopped: 9 times for the long protocol (18.75 %) and 4 times for the short protocol (9.09 %) with statistically significant difference. The number of collected oocytes was significantly higher in the short protocol (7,64±3,70 vs 4,55±2,01, P<0.001). We significantly obtained more embryos in the short protocol (4,31±2,9 vs 2,16±2,2 embryos ; p<0,001). With higher number of grade 1 embryos (2,61 vs 1,14 embryons; p<0.001).The results in terms of pregnancy and living births show no significant difference between the 2 groups. CONCLUSION: The short protocol is more suited to the profile of ovarian poor responders. The long protocol standard has no place in poor responders. However, the long micro dose protocol and the long degressed micro dose protocol yield results at least equivalent to the short protocol.

[Basal fsh level is only predictive of the quantitative aspect of the ovarian response]. / Le taux basal de FSH à j3 du cycle est un facteur prédictif de l'aspect quantitatif de la réponse ovarienne à la stimulation.

BACKGROUND: Determination FSH and LH at day 3 of the menstrual cycle predicts the response to stimulation. AIM: To evaluate the value of FSH and LH measurements compared with women's age in predicting qualitative and quantitative ovarian response to gonadotrophin stimulation. METHODS: 305 patients underwent at least one intra cytoplasmic sperm injection (ICSI) cycle. The levels of FSH and LH at day 3 were determined in an earlier cycle. A good quantitative ovarian response was defined as ³3 oocytes retrieved and 3 embryos obtained. A good qualitative ovarian response was defined as a percentage of mature oocytes ³75% and immature ones²15% of the total number of oocytes retrieved with at least one top quality embryo obtained. RESULTS: Receiver operating characteristic (ROC) curves were generated for FSH, LH and female age. FSH is better than female age in predicting the number of oocytes retrieved (respectively ROCAUC=0.77, p=10-3 versus ROCAUC=0.73, p=10-3) and the number of embryos obtained (ROCAUC=0.69, p=10-3 versus ROCAUC=0.66, p=10-3). LH is non predictive. None of the three tested parameters was predictive of the fertilization and pregnancy rates. An FSH cutoff was calculated and a value of 7.8mUI/ml is associated with a sensitivity of 73% and a specificity of 70% for the prediction of ovarian response to controlled stimulation. CONCLUSION: Basal FSH level predicts good quantitative rather than qualitative response. LH is non predictive. FSH and LH do not predict pregnancy rate. Patients having high FSH levels should not be excluded from IVF/ICSI treatment.

A recurrent deletion of DPY19L2 causes infertility in man by blocking sperm head elongation and acrosome formation.

An increasing number of couples require medical assistance to achieve a pregnancy, and more than 2% of the births in Western countries now result from assisted reproductive technologies. To identify genetic variants responsible for male infertility, we performed a whole-genome SNP scan on patients presenting with total globozoospermia, a primary infertility phenotype characterized by the presence of 100% round acrosomeless spermatozoa in the ejaculate. This strategy allowed us to identify in most patients (15/20) a 200 kb homozygous deletion encompassing only DPY19L2, which is highly expressed in the testis. Although there was no known function for DPY19L2 in humans, previous work indicated that its ortholog in C. elegans is involved in cell polarity. In man, the DPY19L2 region has been described as a copy-number variant (CNV) found to be duplicated and heterozygously deleted in healthy individuals. We show here that the breakpoints of the deletions are located on a highly homologous 28 kb low copy repeat (LCR) sequence present on each side of DPY19L2, indicating that the identified deletions were probably produced by nonallelic homologous recombination (NAHR) between these two regions. We demonstrate that patients with globozoospermia have a homozygous deletion of DPY19L2, thus indicating that DPY19L2 is necessary in men for sperm head elongation and acrosome formation. A molecular diagnosis can now be proposed to affected men; the presence of the deletion confirms the diagnosis of globozoospermia and assigns a poor prognosis for the success of in vitro fertilization.

[Clinicals and biological aspects of ICSI cycles on patients with polycystic ovary syndrome]. / Aspects cliniques et biologiques de l'ICSI chez les femmes porteuses du syndrome des ovaires Polykystiques.

AIM: To analyse clinical et biological pecularities of Polycystics Ovarian Syndrome (PCOS) patients enrolled on ICSI cycles and compare them to normo-ovulatory women. METHODS: 100 controlled ovarian stimulation cycles for ICSI in women with PCOS and 200 cycles in normo-ovulatory women. RESULTS: There was no significant difference in term of cancellation rate (5.5% in PCOS group vs 5%; NS). The mean number of follicles was higher in patients with PCOS (18.1 +/- 8.5 vs 9.4 +/- 5.5; p < 0.05). Oocyte mature rate and fertilization rate were higher in PCOS group (67% vs 52%; p < 0.05) (75% vs 63.7%; p < 0.05) respectively. Grade 1 embryo rate was significantly higher in PCOS group (69% vs 53%; p < 0.05). Implantation rate (16.6% vs 12.1%; NS) and clinical pregnancy rate per transfer (31.5% vs 22.2%; NS) did not differ statistically in the two groups. Miscarriage rate was higher in PCOS group but this did not reach the statistical significance (20% vs 7.1%; NS). 11 cases of Ovarian hyperstimulation syndrome occurred in PCOS group versus 4 on normo-ovulatory group. CONCLUSION: Use of ICSI as fertilization technique was correlated with good biologic parameters on PCOS patients with better fertilization rate and embryo quality and similar pregnancy rate comparing to normo-ovulatory women. However, it still be great concern about high risk of miscarriages and Hyperstimulation ovarian syndrome.

[A retrospective study of 339 ICSI cycles: assesment of the two first years of activity of the Assisted Reproductive Technology Center of Aziza Othmana Hospital]. / Etude retrospective de 339 cycles d'injection intra cytoplasmique de spermatozoïdes: bilan des deux premières années d'activité de l'unité d'assistance médicale à la procréation de l'hôpital Aziza Othmana.

BACKGROUND: Intracytoplasmic sperm injection (ICSI) is a micromanipulation-assisted fertilization, whereby one spermatozoon is injected into the oocyte cytoplasm. Initially, ICSI was the treatment of choice for male factor infertility. However, because of the high fertilization and pregnancy rates achieved with this technique, the scope of the procedure has been widened to include couples with other causes of infertility. AIM: The aim of this study was to study the progression of the activity of the assisted reproductive technology's center of Aziza Othmana's Hospital and the ICSI results during the first two years. METHODS: Our study included 269 infertile couples who underwent 339 ICSI cycles between 1st May 2001 and 30 April 2003. Cycles with no oocytes obtained at the follicular aspiration and women aged over 40 years were excluded from this study. RESULTS: The number of ICSI cycles progressed in our center: 150 ICSI cycles in the 1st year, 189 ICSI cycles in the 2nd year. The mean number of picked-up oocytes was 8,8 +/- 5,6. The fertilization rate was 62%. The mean number of transferred embryos was 3,1 +/- 1,5. The pregnancy rate per transfer was 32,4%. The miscarriage rate was 28,4%. The take home baby rate was 67,9%. CONCLUSION: The number of couples undergoing ICSI cycles in our center is increasing. The fertilization rates and pregnancy rates in our center are similar to those published in the literature.

[Increased risk of early pregnancy loss and lower live birth rate with GNRH antagonist vs. long GNRH agonist protocol in PCOS women undergoing controlled ovarian hyperstimulation]. / Augmentation du taux de fausses couches spontanees precoces et diminution du taux de naissances vivantes avec le protocole antagoniste de la GNRH versus agoniste long chez les patientes porteuses d'un syndrome des ovaires polykystiques.

AIM: to compare standard long GnRH agonist protocol (Triptorelin) and GnRH antagonist regimens (Cetrorelix) in polycystic ovary syndrome (PCOS) patients undergoing controlled ovarian stimulation (COS) for ICSI cycles. METHODS: Retrospective case-control study. 106 PCOS patients undergoing COS for ICSI with long GnRH agonist protocol (Triptorelin) were matched with age and BMI to 106 PCOS patients undergoing COS for ICSI with GnRH antagonist (Cetrorelix) during the same period. Ovarian stimulation with recombinant follicle stimulating hormone (rFSH) was used in the two groups. Oral contraceptive pill pretreatment was used in all patients undergoing ovarian stimulation using GnRH antagonists. ICSI was performed for male infertility in all cases. The main outcome measures evaluated were: cancellation of the cycles, number of aspirated follicles, oocyte maturity, fertilization rate, Embryo quality, pregnancy and implantation rates, clinical abortion rate, multiple pregnancy rate and the live birth rate rate. Kchi2 test and t Student test were used for differences between normo-ovulatory and PCOS patients and the limit of significance was set at p < 0.05. RESULTS: There was no significant difference in term of cancellation rate (2.8% vs 1.8%; NS). Duration of gonadotrophin stimulation (9.7 +/- 0.7 vs. 11.2 +/- 1.9 days; p < 0.001) and gonadotrophin consumption (2209.0 +/- 548.3 vs. 1411.1 +/- 217.9 UI: p < 0.001) were significantly decreased with GnRH antagonist. The mean oestradiol level on the triggering day was significantly higher in the agonist group (3347.85 +/- 99 vs. 2354.45 +/- 839; p < 0.001 ).A fall in LH level of > or = 50% from stimulation day 8 (S8) to S1 was observed in GnRH antagonist group. Risk of ovarian hyperstimulation syndrome (OHSS) was significantly decreased with GnRH antagonist (1.8% vs 10.7%; p = 0.01). The mean number of retrival oocytes (15.9 +/- 5.9 vs. 17.3 +/- 8.3; ns) and the mean number of mature oocytes (11.43 +/- 4.2 vs. 11.91 6.4; ns) were similar in the two groups, fertilization rate (73.3% vs 75.8%; NS), mean number of grade 1 and 2 embryos (6.3 +/- 2.7 vs. 6.9 +/- 3.9; NS), mean number of transferred embryos (1.9 +/- 0.7 vs. 1.8 +/- 0.7; NS), implantation rate (13.3% vs. 18.45%; ns) and clinical pregnancy rate per transfer (28.6% vs 31.1% ; NS) did not differ statistically in the two groups. Twin and triplet pregnancies rates were also similar in the two groups (7.1% vs. 9.3%; NS) and (3.5% vs. 3.1%; NS) respectively. Live birth rate (12.2% vs. 20.7%; p < 0.001) was significantly lower in GnRH antagonist group and miscarrage rate was significantly higher in this same group (42.8% vs. 18.7%; p < 0.001). CONCLUSION: GnRH antagonist protocol is a short and simple protocol with a significant reduction in incidence of OHSS and amount of gonadotrophins. However, GnRH antagonist protocol provides a lower live birth rate and an increased risk of early pregnancy loss compared to the GnRH agonist long protocol. Further studies are necessary for more solid conclusions.