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Percutaneous epididymal sperm aspiration (PESA) in men with obstructive azoospermia.

OBJECTIVES: Assessing the efficiency of repeated percutaneous epididymal sperm aspiration (PESA) in men with obstructive azoospermia, and also the possibility of cryopreservation of remaining material for future use in intracytoplasmic sperm injection (ICSI). METHOD: Retrospective study, in which 79 procedures of PESA were assessed in 58 patients (mean age = 45 years), whose partners had mean age of 34 years. Vasectomy was the most frequent cause of obstructive azoospermia (n = 46). RESULTS: Motile spermatozoa were obtained in 65 procedures (82%). PESA was twice repeated for 15 patients, 3 times for 5 patients, and 4 times for 1 patient. Spermatozoa were found in 13 (87%) patients in the second attempt, in 4 (80%) patients in the third attempt, and in the only patient that had accomplished 4 procedures. In 30 procedures (37%), we have obtained enough material for cryopreservation. In 12 among the 13 samples thawed (n = 13 patients), motile spermatozoa were found, and ICSI was accomplished. Four patients that did not use their samples requested the elimination of the material. Total rate of pregnancy per transference was 21/55 (38%). In 14 PESA procedures, it was not possible to find spermatozoa; in these cases, the patients opted for accomplishing the procedure of testicular sperm aspiration (TESA). CONCLUSION: PESA is an efficient and simple method of retrieving spermatozoa, allowing repeated procedures. Additionally, spermatozoa collected through PESA can be cryopreserved.

Journal Article↗

Zona-free oocyte fertilized with intracytoplasmic sperm injection and underwent further division: case report and literature review.

The zona pellucida (ZP) plays a protective role during fertilization and early embryonic development. It is related to sperm binding, the acrosome reaction, prevention of polyspermic fertilization, and holding blastomeres together before the morular stage. Zona-free oocytes are accidentally encountered. If these oocytes are healthy, they can be fertilized normally by intracytoplasmic sperm injection (ICSI). We reported on a couple with male infertility undergoing oocyte retrieval after ovarian hyperstimulation. Before the ICSI procedure, cumulus cells surrounding the oocytes were removed, which resulted in one oocyte escaping from its ZP. The zona-free oocyte was fertilized normally with ICSI and developed to the 8-cell stage. We observed that the zona-free zygote had the ability to further divide, despite its loose contact. The zona-free embryo was transferred with other zona-intact embryos, but the implantation failed. We conclude that zona-free oocytes can be rescued, fertilized with ICSI, and cultured for further transfer or cryopreservation.

Adult↗

Efficacy of microsurgical epididymal sperm aspiration (MESA) and intracytoplasmic sperm injection (ICSI) in obstructive azoospermia.

PURPOSE: Our purpose was to test whether micromanipulation using subzonal insemination and intracytoplasmic sperm injection could improve the poor fertilization and pregnancy rates obtained when attempting in vitro fertilization in patients with congenital absence of the vas deferens and unreconstructable obstructive azoospermia with microsurgically retrieved epididymal spermatozoa. RESULTS: Conventional in vitro fertilization (group A; 14 cycles), subzonal insemination (group B; 13 cycles), and intracytoplasmic sperm injection (group C; 28 cycles) were carried out in 55 treatment cycles. Fertilization rates for groups A, B, and C were 16.1, 31.4, and 48.6%, respectively (P < 0.05). Clinical pregnancy rates for groups A, B, and C were 7.1, 7.7, and 32.1% (P < 0.05), respectively. In five cycles, intracytoplasmic sperm injection using epididymal sperm from alloplastic spermatoceles was performed and two clinical pregnancies (40%) were obtained. CONCLUSIONS: The combined microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection procedure is highly effective in improving the fertilization and pregnancy rate in congenital absence of the vas deferens and unreconstructable obstructive azoospermia. Furthermore, alloplastic spermatoceles may be useful for repeat sperm aspirations.

Epididymis↗

Intracytoplasmic sperm injection increased fertilization and good-quality embryo formation in patients with non-male factor indications for in vitro fertilization: a prospective randomized study.

OBJECTIVE: To compare the fertilization rate and formation of good-quality embryos with conventional IVF and ICSI in patients with non-male factor infertility. DESIGN: Prospective controlled study. SETTING: Infertility clinic. PATIENT(S): Thirty-five patients with non-male factor infertility. INTERVENTION(S): Retrieved sibling oocytes were randomly assigned to conventional IVF or ICSI. Of sibling oocytes assigned to ICSI, only metaphase II oocytes were injected with sperm. MAIN OUTCOME MEASURE(S): Fertilization rate and formation of good-quality embryos per retrieved oocyte. RESULT(S): Per retrieved oocyte, ICSI resulted in better fertilization rate compared with conventional IVF (71.3% [134 of 188] vs. 57.2% [107 of 187]). Per retrieved oocyte, ICSI also resulted in better formation of good-quality embryos at 48 hours after retrieval compared with conventional IVF (64.4% [121 of 188] vs. 47.1% [88 of 187]). CONCLUSION(S): In IVF patients with non-male factor infertility, subjecting some sibling oocytes to ICSI increased the fertilization rate and formation of good-quality embryos per retrieved oocyte. It also avoided the problem of total fertilization failure in almost all cases.

Adult↗

Ovarian tissue banking for cancer patients: is ovarian cortex cryopreservation presently justified?

The effect of chemotherapy and radiotherapy on future fertility is of concern to patients and their families. Whereas sperm banking is commonly performed, female gametes are not so amenable to cryopreservation. One alternative includes postponing cancer treatment to enable ovulation induction and oocyte aspiration. Whenever possible, retrieved oocytes should be fertilized in vitro prior to cryopreservation. Frozen embryos could serve to produce pregnancies if ovarian failure occurs. Donor sperm can be offered to single patients, as frozen-thawed unfertilized oocytes yield poor pregnancy rates. Ovarian cortex cryopreservation should still be considered an experimental technology as no pregnancies have been obtained in humans. Therefore, ovarian cortex banking should be used only for young girls, adolescents and when IVF is contraindicated. Reattachment of ovarian vasculature could prevent ischaemic follicular loss and enable ovarian transplantation in the future. This procedure is currently under investigation in animals. At the present time, we recommend urgent IVF in most patients requesting fertility preservation. Ovarian cryopreservation should be offered when emergency IVF is not possible.

Animals↗

Sperm characteristics and outcome of human assisted fertilization by subzonal insemination and intracytoplasmic sperm injection.

OBJECTIVE: To investigate the influence of sperm characteristics on the treatment by subzonal insemination (SUZI) and intracytoplasmic sperm injection of couples with severe male infertility. DESIGN: A retrospective analysis of 300 consecutive cycles of assisted fertilization concerning 202 infertile couples was performed. One hundred fifty-three couples underwent 362 unsuccessful IVF cycles, whereas on 49 couples IVF was not performed because of poor sperm characteristics. SETTING: Procedures were performed in an institutional research environment. PATIENTS, PARTICIPANTS: Couples in which the male partner was the presumed cause of repeated failure to achieve conception by IVF or in which seminal parameters were unacceptable for IVF. INTERVENTIONS: Three hundred transvaginal oocyte retrievals were performed after superovulation by GnRH agonist and gonadotropins. MAIN OUTCOME MEASURES: After SUZI and intracytoplasmic sperm injection the following parameters were evaluated: fertilization, cleavage, pregnancy, and implantation rates in relation to the sperm parameters and the proportion of acrosome-free spermatozoa after different treatments. RESULTS: Normal fertilization occurred in 18% of the oocytes treated by SUZI and in 44% after intracytoplasmic sperm injection. Only the treatment by electroporation showed a positive correlation with the fertilization rate. Fourteen pregnancies were obtained after SUZI, 8 pregnancies after intracytoplasmic sperm injection, and 8 pregnancies after a combination of the two procedures. A score calculated from the sperm parameters after selection correlated with the fertilization obtained after SUZI, whereas a score calculated from the parameters before sperm selection correlated with the pregnancy rate. Sperm morphology influenced the implantation rate of the embryos obtained with these two procedures. CONCLUSIONS: Intracytoplasmic sperm injection and SUZI can successfully treat couples who fail IVF or who cannot benefit from IVF. Different treatments can be applied to semen samples to increase the number of acrosome-reacted spermatozoa. The few significant relations found between sperm characteristics and the outcome of assisted fertilization cannot predict the outcome.

Adult↗

Possible factors affecting the development of oocytes in in-vitro maturation.

To date, pregnancy rates from oocytes matured in-vitro (IVM) have been much lower than those with in-vivo stimulated maturation. In order to improve the developmental potential of IVM oocytes, we studied the effect of three possible factors on pregnancy rates: (i) priming in vivo with FSH before aspiration; (ii) the time interval of maturation in vitro, and (iii) timing the aspiration by monitoring the serum concentrations of oestradiol and inhibin A. In all experiments, oocyte retrieval was performed transvaginally and oocytes were matured individually in culture medium (TCM 199) under oil. Intracytoplasmic sperm injection (ICSI) was carried out on all metaphase II oocytes. Suitable embryos (maximum of two) were replaced after culturing for 2-3 days in IVF medium. Endometrial priming consisted of 2 mg 17beta-oestradiol, taken orally three times a day from the day of oocyte retrieval, and intravaginal progesterone suppositories initiated 2 days later. In the first experiment, 20 women were randomly allocated to two groups: group I (n = 10 cycles) received no stimulation, while group II (n = 10 cycles) received recombinant FSH 150 IU/day for 3 days, initiated on day 3. FSH priming did not affect the rates of maturation, fertilization or cleavage, and no effect was seen on embryo development. The second experiment included 48 patients undergoing 55 unstimulated cycles. The effects of IVM periods of 28 and 36 h were compared. Shortening the IVM period did not compromise subsequent embryo development. The third study analysed the results of maturation of oocytes obtained in 87 cycles in 75 unstimulated normal women, after a leading follicle of 10 mm in diameter and an endometrial thickness of at least 5 mm were observed. A pregnancy rate of 12.6% (11/87) per aspiration and 17.4% (11/63) per transfer was obtained. Serum concentrations of oestradiol and inhibin A were evaluated retrospectively. Significantly more pregnancies were obtained in cycles with a detected increase in the concentration of oestradiol from day 3 to the day of aspiration (19% per aspiration) compared with cycles without such an increase (0% per aspiration). A higher pregnancy rate was observed after an increase in inhibin A concentration (24 versus 0%). In conclusion, monitoring the timing of aspiration may potentially improve the developmental potential of immature oocytes. FSH priming did not affect either cleavage rate or embryo development. Shortening the maturation period from 36 to 28 h did not compromise subsequent embryonic development.

Adolescent↗

Delivery after transfer of frozen-thawed embryos from in vitro-matured oocytes in a woman at risk for ovarian hyperstimulation syndrome.

The present report describes the birth of a healthy infant after cryopreservation of embryos produced from in vitro-matured oocytes retrieved from a woman at risk of developing ovarian hyperstimulation syndrome (OHSS) during conventional in vitro fertilization (IVF) cycles. A conventional long protocol including gonadotropin-releasing hormone agonist (GnRHa) and gonadotropins induced a risk of OHSS. Oocyte retrieval was performed on day 11 of the cycle, and 27 immature oocytes were obtained. Following incubation for 24 h in maturation medium, 74.1% (20/27) of the oocytes were at the metaphase II stage. Fourteen oocytes (14/20, 70.0%) were fertilized after intracytoplasmic sperm injection (ICSI) with her husband's spermatozoa and cultured for 3 days. On day 4 following oocyte retrieval, three embryos at the 8-16 cell stage were transferred into the woman's uterus, and five spare embryos were frozen. Since the fresh embryo transfer failed to result in pregnancy, three post-thaw embryos were transferred into the woman three months later. Transfer of the frozen embryos resulted in pregnancy with delivery of a healthy infant girl.

Adult↗

Births of kittens produced by intracytoplasmic sperm injection of domestic cat oocytes matured in vitro.

In Experiment 1, cleavage frequency and in vitro development of domestic cat embryos produced after in vitro maturation of oocytes obtained from ovaries after ovariohysterectomy (in vitro) with that of oocytes retrieved from follicle-stimulating hormone-treated donors at 24 h after administration of luteinizing hormone (in vivo) and fertilization by intracytoplasmic sperm injection (ICSI) or IVF were compared. In each group presumptive zygotes were assessed for cleavage on IVC Days 1 and 4 and for development to blastocysts on IVC Day 7. In vitro matured oocytes had lower frequencies of meiotic maturation (59.2% v. 66.5%), cleavage at Day 1 (41.4% v. 64.9%) and development to the morula stage at Day 4 (65.8% v. 87.9%) than did in vivo matured oocytes, after ICSI and IVF. Development to the blastocyst stage was lower in in vitro matured oocytes (19.0%) than in vivo matured oocytes (29.5%) after ICSI. In Experiment 2, we evaluated the capacity of sperm injected oocytes without a visible polar body to undergo cleavage and in vitro development. More in vivo matured than in vitro matured oocytes underwent cleavage at Day 1 (46.6% v. 12.6%) and developed to the morula stage by Day 4 (66.7% v. 46.1%), but no blastocysts were obtained at Day 7 in either group. In Experiment 3, we evaluated the in vivo viability of domestic cat embryos derived from ICSI of in vitro matured oocytes. Morula stage embryos were transferred to 18 domestic cat recipients either on Day 4 or 5 after oocyte recovery. A total of 3 domestic cat recipients were pregnant after transfer to recipients on Day 5. Two pregnant cats delivered two normal and healthy live male kittens on Day 68 of gestation and the remaining cat delivered a male kitten on Day 62 that died during the last two days of gestation. These results demonstrate that: (1) inadequate cytoplasmic maturation of in vitro matured domestic cat oocytes is the main cause of deficient oocyte activation; (2) the injection of oocytes without a visible polar body is a useful technique to evaluate oocyte cytoplasmic maturation; and (3) blastocysts obtained after ICSI of in vitro matured oocytes are viable and not a result of parthenogenesis.

Animals↗

Laser-assisted cryopreservation of single human spermatozoa in cell-free zona pellucida.

Improved procedure for efficient cryopreservation of single human spermatozoa in cell-free human zona pellucida is reported. We used a diode laser system for efficient and precise creation of a single hole into the zona pellucida of a degenerated or immature human oocyte. This allowed the extraction of the cytoplasm using a micropipette with a diameter of 10-15 microns. Through the same opening, human spermatozoa were inserted into the empty zona. We used motile and laser immobilized spermatozoa. Immobilized sperm were obtained by a single laser irradiation delivered in the vicinity of the sperm tail prior to insertion. This new immobilization procedure was shown to have no deleterious effect on membrane integrity and sperm viability. Following sperm transfer into the zona, the laser-drilled hole was closed with an oil droplet which was expelled from the micropipette during withdrawal to avoid loss of spermatozoa. This facilitated detection of the otherwise translucent zona during the cryopreservation procedure. After thawing, all cryopreserved zonae (20/20) could be successfully retrieved. Spermatozoa were recovered from the zona pellucida through the hole used for insertion. The rate of sperm recovery for initially motile spermatozoa was 80% vs. 92% for laser immobilized spermatozoa. Sperm viability was 81% and 84%, respectively, detected by a Hoechst stain. This technique makes cryopreservation of single human spermatozoa easy and feasible and appears beneficial for couples with severe male infertility and for those facing repeated surgical sperm extraction.

Cryopreservation↗

Performance of different ovarian reserve markers for predicting the numbers of oocytes retrieved and mature oocytes.

OBJECTIVE: To assess the performance of different ovarian reserve tests for predicting the total numbers of oocytes retrieved and mature oocytes. METHODS: A retrospective study was performed on 71 women undergoing their first IVF/intracytoplasmic sperm injection treatment. Basal ovarian reserve screening was performed on days 2-3 of a spontaneous cycle. Patients were down-regulated with the GnRH agonist, whereas ovarian stimulation was carried out with recombinant FSH, starting from day 2 to 3 of the cycle. The main outcome measures were the numbers of oocytes retrieved and mature oocytes. RESULTS: The total number of oocytes was positively correlated with AFC (p<0.0001) and E(2) levels post-GnRH (p<0.004), whereas there was an inverse correlation with age (p<0.0001). The number of mature oocytes also correlated with AFC (p<0.008) and E(2) levels post-GnRH (p<0.009), and inversely with age (p<0.0004). Univariate linear regression of square root of number of oocytes (SQNO) shows that acceptable predictors of number oocytes, based on model significance and R(2) are AFC (R(2)=0.215), age (R(2)=0.24) and E(2) variation (R(2)=0.09). The following model is proposed to predict the number of mature oocytes: ln(MO)=ln(NO)-2.09+0.028 AGE+0.03 BMI. CONCLUSION(S): The number of antral follicles and patient age appear to be good markers of ovarian response in IVF treatments. Additional information is provided by stimulated E(2) levels.

Adult↗

Epididymal sperm aspiration with assisted reproductive techniques: difference between congenital and acquired obstructive azoospermia?

From July 1992 to May 1994, 31 cases of obstructive azoospermia-induced infertility underwent 35 epididymal sperm aspiration procedures and assisted reproductive techniques. These included in-vitro fertilization (IVF), zygote intra-Fallopian transfer (ZIFT), subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI) with embryo transfer or tubal embryos transfer. The motile spermatozoa were retrieved using a gauge 24 Medicut cannula and flush medium. Total oocyte fertilization rate was 23.4% and the clinical pregnancy rate was 12.5% (four out of 32 treatment cycles). Congenital absence of the vas deferens was found in 16 cases (51.6%) and secondary genital duct obstruction was present in 15 cases (48.4%). In all, 29 aspirations were retrieved from the caput of the epididymis and six from the epididymal body or tail. The fertilization rate for the caput spermatozoa was much less than that for other areas of the epididymis (P < 0.05). Though there were no predictable differences in fertilization rates between the congenital and acquired groups, by using epididymal spermatozoa for assisted reproduction, the congenital group seemed to have a stronger tendency to achieve pregnancy (20 versus 5.9%).

Adult↗

Changes in motility patterns during in-vitro culture of fresh and frozen/thawed testicular and epididymal spermatozoa: implications for planning treatment by intracytoplasmic sperm injection.

The present report describes the motility changes in vitro (percentage motile and progressively motile) of freshly collected testicular and epididymal spermatozoa and following freeze/thaw of the same spermatozoa from a man with obstructive azoospermia. Washed spermatozoa were cultured in micro droplets under paraffin oil or in test tubes using HEPES-buffered or bicarbonate-buffered medium containing 10% human serum. In fresh testicular sperm cultures 60-65% of the sperm cells became motile within 2 days of culture; the motility was maintained for a further 4-5 days before a decline was observed. The progressive motility improved markedly on the third day of culture and it peaked around day 5. Only a small number of frozen/thawed testicular spermatozoa became motile during in-vitro culture (15-20%) and the motility was maintained for only 2-3 days before it declined. Furthermore, only 10-12% of the spermatozoa showed progressive motility. Spermatozoa recovered from micro-epididymal sperm aspiration (MESA) showed a gradual decrease in progressive motility and in 5 days all sperm cells were found to be immotile in both freshly collected and frozen/thawed spermatozoa. All culture systems supported sperm motility. It is clear that testicular spermatozoa, particularly from men with obstructive azoospermia, can be collected and maintained in vitro for up to 1 week before the oocyte retrieval but when frozen testicular or epididymal spermatozoa are used it is more reliable to thaw these spermatozoa on the day of intracytoplasmic sperm injection.

Adult↗

A prospective, randomized, double-blind clinical trial to study the efficacy and efficiency of a fixed dose of recombinant follicle stimulating hormone (Puregon) in women undergoing ovarian stimulation.

A prospective, randomized, double-blind, multicentre (n = 5) study was conducted to compare the influence of either a 100 or 200 IU daily fixed-dose regimen of recombinant follicle stimulating hormone (FSH) on the number of oocytes retrieved and the total dose used in down-regulated women undergoing ovarian stimulation. Fertilization was done by intracytoplasmic sperm injection or conventional in-vitro fertilization. A total of 199 women were treated with FSH, 101 subjects with 100 IU and 98 subjects with 200 IU. In subjects of the 200 IU treatment group, significantly more oocytes were retrieved compared to the 100 IU group (10.6 versus 6.2 oocytes, P < 0.001). The total dose needed to develop at least three follicles with a diameter of > or = 17 mm was significantly lower in the 100 IU treatment group (1114 IU versus 1931 IU, P < 0.001). In the low-dose group, significantly lower serum concentrations of oestradiol, progesterone and FSH were observed at the day of human chorionic gonadotrophin administration. Although more cycle cancellations due to low response were seen in the 100 IU group (n = 24 versus n = 3), the clinical pregnancy rate per started cycle was similar (24.7% in the 100 IU group versus 23.3% in the 200 IU group). In the high-dose group, more side-effects, in particular more cases of ovarian hyperstimulation syndrome, were noted. It is concluded that compared to 200 IU, the use of a 100 IU fixed dose is less efficacious in terms of the number of oocytes retrieved, but more efficient as indicated by a lower total dose.

Buserelin↗

Efficacy and clinical outcome of preimplantation genetic diagnosis using FISH for couples of reciprocal and Robertsonian translocations: the Korean experience.

OBJECTIVE: To evaluate the efficacy and clinical outcome of preimplantation genetic diagnosis (PGD) using fluorescence in situ hybridization (FISH) for couples with chromosomal translocations. METHODS: PGD using FISH was performed in 59 cycles of 43 couples with reciprocal translocations, and 11 cycles of 6 couples with Robertsonian translocations. The diagnostic and clinical data were reviewed in a series of 70 treatment cycles of 49 couples from January 2001 to June 2002 at Samsung Cheil Hospital, Korea. RESULTS: A total of 1408 oocytes were retrieved, and 938 (81.7%) out of 1148 matured oocytes were fertilized by intracytoplasmic sperm injection (ICSI). Single blastomere biopsy and FISH analysis were successfully carried out in 99.3% (890/896) and 94.4% (840/890), respectively. Among 193 normal or balanced embryos, 169 embryos were transferred in 64 cycles (91.4% per started cycle). Twenty clinical pregnancies including two ectopic pregnancies and three spontaneous miscarriages (28.6% per started cycle, 31.3% per transfer cycle, 40.8% per couple) were established. Of the three spontaneous miscarriages, one was karyotyped as normal, one had an unbalanced arrangement and one was tetraploid. One case of preterm twin delivery occurred and 16 healthy babies were delivered in 12 single and 2 twin pregnancies. CONCLUSION: The clinical outcome was successful in 28.6% (14/49) of the treated couples with translocations after PGD. The spontaneous abortion rate was significantly reduced from 95.8% (69/72) to 16.7% (3/18) in these couples.

Abortion, Spontaneous↗

Normal infant by a gestational carrier for a phenylketonuria mother: alternative therapy.

The consequences of pregnancies in untreated phenylketonuria (PKU) mothers are a high incidence of spontaneous abortion, intrauterine growth retardation with microcephaly, congenital malformations, and abnormal intellectual development. PKU fathers, on the other hand, produce normal children. Obviously children of PKU women and men are at least heterozygous, proving that the abnormalities produced by the PKU mothers are not genetic but "intrauterinely environmental." Exposure to the mother's metabolic abnormalities affects the fetus during the entire pregnancy. A PKU mother can produce a healthy infant if she maintains a very restricted and controlled diet before and during pregnancy. However, even the most recent reports describe a very high incidence of congenitally abnormal children of PKU mothers, hence dietary compliance is not working in all cases. A 26-year-old PKU patient with proven fertility underwent standard ovarian stimulation in preparation for oocyte retrieval. Following conventional co-incubation of the oocytes and her husband's sperm, two embryos were transferred to the gestational carrier's uterine cavity, resulting in a single intrauterine pregnancy. Birth was induced at 39 weeks of gestation. The male infant weighed 3486 g. Head circumference was 36 cm and length 50.5 cm; there was no evidence of any abnormality and/or malformation. At 1 year of age, the child's growth measurements and development assessments were normal. This describes the first reported successful term pregnancy of an untreated PKU mother with the help of a gestational carrier (GC), producing a normal infant. This is an alternative method that should be offered to PKU women who are unable and/or unwilling to maintain a well controlled diet before and during pregnancy.

Complementary Therapies↗

Reproducibility of transvaginal three-dimensional endometrial volume measurements with virtual organ computer-aided analysis (VOCAL) during ovarian stimulation.

OBJECTIVE: To calculate inter- and intraobserver reliabilities for three-dimensional endometrial volume measurements during in vitro fertilization using virtual organ computer-aided analysis (VOCAL). PATIENTS AND METHODS: Three-dimensional ultrasound measurements of the endometrium using VOCAL software were performed on the day of oocyte retrieval in each of 79 consecutive patients undergoing in vitro fertilization or intracytoplasmic sperm injection. Endometrial volume was calculated every 15 degrees and every 30 degrees by two different observers in order to determine the reproducibility of the technique. RESULTS: Intraobserver reliability for the 15 degrees measurements was 0.97 for both observers; for the 30 degrees measurements, it was 0.93 for one observer and 0.96 for the other. Interobserver reliability was 0.80 for the endometrial volumes calculated every 15 degrees and 0.83 for the volumes calculated every 30 degrees. The intra- and interobserver measurement agreement showed good reproducibility. However, the volumes calculated every 15 degrees were more accurate because the means of differences were close to zero. CONCLUSION: VOCAL provides a reproducible method for the estimation of the endometrial volume.

Adult↗

Reduced aromatase activity in granulosa cells of women with endometriosis undergoing assisted reproduction techniques.

The aim of the present study was to measure the in vitro aromatase activity in granulosa cells of women with endometriosis submitted to assisted reproduction techniques. A case-control study was conducted on eight patients with endometriosis and eight with other infertility causes submitted to in vitro fertilization or intracytoplasmic sperm injection. Granulosa cells were obtained from pre-ovulatory follicles during oocyte retrieval and cultured for 24 h in the presence or absence of testosterone (2 x 10(-6) and 2 x 10(-5) M), follicle-stimulating hormone (FSH) and insulin-like growth factor-I (IGF-I) (both at 50 ng/ml). Estradiol (radioimmunoassay) was measured in the obtained culture fluids. The basal production of estradiol and its production under testosterone addition to the culture (aromatase activity) were analyzed. Reduced aromatase activity was detected in cultured granulosa cells in endometriosis cases, compared with controls, when testosterone was added at the concentration at 2 x 10(-6) M (p = 0.0303). The basal production of estradiol was also reduced in endometriosis patients (p = 0.0390). The effect of addition of FSH and IGF-I did not differ between groups. In conclusion, the in vitro basal production of estradiol and aromatase activity in granulosa cells were reduced in women with endometriosis submitted to assisted reproduction techniques, compared with the control group.

Adult↗