José G. Franco

Power Doppler Endometrial Evaluation as a Method for the Prognosis of Embryo Implantation in an ICSI Program

AbstractPurpose: The objective of the present study was to evaluatepower Doppler of the endometrium as a parameter for theprognosis of embryo implantation in patients who underwentintracytoplasmic sperm injection (ICSI). Methods: The power Doppler was performed on a transversesection at the level of the uterine fundus on the day of humanchorionic gonadotropin in 185 patients who submitted toovarian stimulation for ICSI. The endometrium was dividedinto four equal quadrants and classified as grade I, II, III,or IV according to the visualization of the power Dopplerin the quadrants. The color Doppler signal was consideredto be positive when it reached at least the basal layer ofthe endometrium. Results: Age, number of days of stimulation, number offollicles ≥ 16 mm, number of oocytes in metaphase IIretrieved, and fertilization rate did not differ patients withthe four different types of endometrial grades. Endometrialthickness and the pulsatility index of uterine artery alsowere similar for the four grades. The rate of embryo implantationalso did not differ significantly (P = 0.53) amonggroups: grade I = 10%; grade II = 11.6%; grade III =15.4%; grade IV = 10.5%. The pregnancy rates were gradeI = 25%; grade II = 29.7%; grade III = 37.5%; gradeIV= 23.8% (P = 0.44). Conclusions: Our data demonstrate that isolated evaluationof endometrial vascularization with power Doppler is notan important factor for the prediction of pregnancy in anICSI program.

Effects of Vaginal Progesterone Administration Starting on the Day of Oocyte Retrieval on Pregnancy Rates

AbstractPurpose: Vaginal progesterone administration starting on the day of oocyte retrieval induced a decrease in uterine contraction frequency on the day of embryo transfer (ET) as compared with preovulatory values. Uterine relaxation before ET is likely to improve outcome by avoiding displacement of the embryo from the uterine cavity (Fanchin, Righini, de Ziegler, Oliviennes, Ledée, Frydman: Fertil Steril 2001;75:1136–1140). The objective of the present study was to determine whether the early use of vaginal progesterone on the day of oocyte retrieval may alter the embryo implantation and pregnancy rates. Methods: A total of 103 patients were submitted for ovarian stimulation with GnRH-a and recombinant FSH (Puregon, Organon) for the application of invasive assisted reproduction techniques (ICSI). The patients were divided into two groups in a prospective and randomized manner: Group A (n = 51) where application of vaginal progesterone started (Utrogestan, Besins International) at the dose of 400 mg from the evening of the day of oocyte retrieval, and Group B (n = 52) started to apply vaginal progesterone at the same dose but from the evening of embryo transfer (2nd day). Results: The age of Group A patients (34.2 ± 4.6) was similar (p = 0.50) to that of Group B patients (34.8 ± 4.9). The number of oocytes retrieved and at metaphase II from Group A patients (10.6 ± 6.9 and 7.8 ± 6.0; respectively) did not differ significantly (p = 0.84 and p = 0.49, respectively) from the number of oocytes retrieved and metaphase II from Group B patients (10 ± 5.6 and 6.7 ± 4.7, respectively). Also, there was no difference (p = 0.48) in number of embryos transferred to Group A patients (2.7 ± 0.8) versus Group B patients (2.7 ± 0.9). Embryo implantation and pregnancy rates for Group A patients (12.6 and 27.4%, respectively) were equal (p = 0.98 and p = 1.0, respectively) to those for Group B patients (13.4 and 28.8%, respectively). Conclusion: Vaginal progesterone at the dose of 400 mg started on the day of oocyte retrieval did not increase implantation or pregnancy rates when compared to the same dose started on the day of embryo transfer.

Laser-assisted hatching of cryopreserved—thawed embryos by thinning one quarter of the zona

Laser-assisted hatching is little documented in the literature regarding its efficacy in cryopreserved-thawed (CT) embryo transfer cycles. The aim of the present study was to evaluate in a randomized manner the efficacy of thinning one quarter of the zona pellucida of CT embryos to a depth of 50-80% of the original thickness, via laser treatment (the qLZT-AH procedure), in improving implantation and pregnancy rates. Two populations were studied: population I, patients who had all their supernumerary embryos cryopreserved, regardless of their morphology, and population II, patients at risk of ovarian hyperstimulation syndrome who had all their embryos cryopreserved. Artificial and natural protocols were used for the embryo transfers. A total of 350 laser-thinned CT embryos were compared with 352 intact zona embryos. No difference in implantation or pregnancy rate was found after using qLZT-AH in either population. These findings suggest that qLZT-AH should not be routinely performed in cryopreserved embryo programmes.

Angiotensin-converting enzyme: Serum levels during normal pregnancy

ACE activity of the serum of 52 normal pregnant women was measured in vitro under conditions of substrate saturation with Hip-His-Leu as substrate. The product His-Leu was measured by fluorimetry after reaction with o-phthaldehyde. ACE activity (nmol/min/ml serum) was 30.6 +/- 7.8, 28.8 +/- 7.4, and 30.9 +/- 8.2 for the first, second, and third trimester of pregnancy, respectively. No statistically significant differences (p greater than 0.05) in ACE activity were detected among the three trimesters of normal pregnancy with either serum volume or serum protein as reference value. These values are within the range reported by Friedland and Silverstein13 for 51 male and seven female healthy blood bank donors. We conclude that the evolution of normal pregnancy does not significantly modify the levels of ACE in peripheral blood serum.

A Prospective, Randomized Comparison of Two Commercial Media for ICSI and Embryo Culture

AbstractPurpose:The aim of this prospective, randomized study was to compare the results obtained in ICSI with two culture media, P-1 (Irvine Scientific) and IVF-50 (Scandinavian IVF Science). Methods: A total of 182 patients undergoing ICSI treatment were randomly included in this study and divided in two groups: Group I: P-1 medium (n = 91) or Group II: IVF-50 medium (n = 91). All the embryos were transferred on the second day. Results: Patient age did not differ (p = .29) between Group I (34.8 ± 4.8) and Group II (34.0 ± 4.5). The number of oocytes retrieved from Group I (10.6 ± 6.7) was also similar (p = .49) to that retrieved from Group II (11.1 ± 6.4). In addition, there was no difference (p = .25) in the number of oocytes retrieved at metaphase II between Group I (7.9 ± 4.6) and Group II (8.7 ± 4.6). Normal fertilization rates, abnormal fertilization rates, and cleavage rates were similar (p = .62, p = .48, and p = .9, respectively) between Group I (68.4 ± 23.3%, 6.7 ± 10.3%, and 98.7 ±4.6%) and Group II (65.3 ± 26.2%, 9.0 ± 13.8%, and 98.9 ± 3.9%, respectively). The embryo score was also similar (p = .62) for both groups (Group I: 31.9 ± 14.0 and Group II: 33.4 ± 15.8). There was no difference in the number of embryos transferred (p = .69) between Group I (2.8 ± 1.0) and Group II (2.8 ± 1.1). In addition, pregnancy rates/puncture, pregnancy rates/transfer, implantation rates, and abortion rates were also similar for Group I (36.2%, 37.0%, 17.4%, and 12.1%, respectively) and Group II (31.8%, 33.7%, 15.8%, and 10.3%, respectively) (p = .64, p = .75, p = .72, and p = 1.0, respectively). Conclusions: There were no differences in the results obtained with culture media P-1 (Irvine Scientific) and IVF-50 (Scandinavian IVF Science) for ICSI and embryo culture.

Administration of β2-Adrenergic Agonists During the Peri-Implantation Period Does Not Improve Implantation or Pregnancy Rates in Intracytoplasmic Sperm Injection (ICSI) Cycles

AbstractPurpose: The objective of the present investigation was to determine implantation and pregnancy rates in patients undergoing ICSI and treated with β2-adrenergic agonists, considering the uterine-relaxing action of these agents. Methods: A total of 225 women undergoing ICSI at the Center for Human Reproduction, “Sinhá Junqueira” Maternity Foundation, entered the study. Patient participation in each group was random, by drawing lots, using a randomization table previously elaborated for the study (2:2:1). The group I (90 women) received 10 mg of terbutaline daily for 15 days starting on the day of oocyte retrieval; group II (90 women) received 20 mg of ritodrine daily during the same period of time as group I; group III (45 patients) received no treatment and was used as control. The evaluation was interrupted in 3 patients of group I and in 30 patients of group II because of a high incidence of side effects. Results: Pregnancy, implantation, and miscarriage rates were not significantly different (p > 0.05) between the three groups: 29.88%, 13.25%, and 26.9% for group I; 33.33%, 17.5%, and 10.0% for group II; 28.88%, 15.07%, and 15.38% for group III, respectively. Conclusions: The results of this study do not support the routine use of β2-adrenergic agonists during the peri-implantation period in assisted reproductive technology cycles.

Psychological Evaluation Test After the Use of Assisted Reproduction Techniques

AbstractPurpose: The emotional changes provoked by the use of assisted reproduction techniques (ART) may trigger important psychological reactions. The objective of the present study was to develop a psychological evaluation test (PET–ART) in order to identify the occurrence of psychological problems and to facilitate their treatment. Methods: A total of 128 women were submitted to PET–ART of the Center for Human Reproduction, “Sinhá Junqueira” Maternity Foundation, after application of IVF/ICSI program at least once. The causes of infertility were male-related in 45% of cases, female-related in 48%, and both male- and female-related in 7%. Infertility was primary in 79% of cases and secondary in 21%. The mean age of the women was 34.5 ± 5.2 years and the mean age of the men was 37.9 ± 6.8 years. The PET–ART was evaluated using a questionnaire with 15 questions selected in order to detect emotional reactions caused by infertility. The responses were assigned four grades with respect to intensity (1 = mild intensity; 2 = medium intensity; 3 = high intensity; and 4 = maximum or unbearable intensity). The sum of the responses corresponded to PET–ART score ranging from 15 to 60 points. The reliability of the questionnaire was evaluated by the alpha coefficient of Cronbach. Results: The PET–ART identified five questions receiving 50% or more responses of the high/maximum intensity type (sum of the percentages of responses with a score of 3 and 4). The questions were the following: 1—The waiting time before being submitted to a pregnancy test (82.8% of the patients); 2—A negative result of the pregnancy test (77.3% of the patients); 3—The degree of anxiety in a new attempt to obtain pregnancy (76.5% of the patients); 4—Finding the money necessary for the repetition of the IVF/ICSI techniques (66.4% of the patients); 5—The possibility of collecting few eggs, or of forming or not an embryo in the laboratory is an expectation that makes me anxious (57.8% of the patients). The mean PET–ART was 33 ± 6. The alpha coefficient of Cronbach was 0.757. Conclusions: The PET–ART was an efficient tool for the identification of women with emotional changes provoked by the application of ART and for the planning of their treatment. However, a general psychological approach was developed for each emotional factor regardless of PET–ART.

Constriction of Human Umbilical Artery Induced by Local Anesthetics

In order to study the direct effects of local anesthetics on the umbilical artery, an in vitro perfusion technique, utilizing 3-cm segments of human umbilical artery obtained within 15 min of delivery, has been developed. Basal perfusion pressure (tonus) and the frequency of phasic contractions were determined in 37 arterial segments perfused for 240-min periods with Tyrodes solution containing lidocaine in concentrations of 2.0, 8.0 or 16 microgram/ml of bupivacaine in concentrations of 0.25 or 9.8 microgram/ml. A control group consisting of 11 arterial segments was perfused only with Tyrodes solution. The lowest concentration of each anesthetic approximated that reportedly present in the umbilical arteries following administration of epidural anesthesia during labor. Both lidocaine and bupivacaine induced dose-related increases of up to 100% in tonus above the control levels of approximately 60 mm Hg, statistically significant (p less than 0.05) except for 2 microgram/ml lidocaine. At the same molar concentration, bupivacaine was more effective than lidocaine (p less than 0.05). Both anesthetics also increased the frequency of phasic contractions. These data suggest that these agents may exert their effects on the fetus, at least in part, through a direct constricting action on the umbilical artery.

The effects of age on sperm quality: an evaluation of 1,500 semen samples

Objective: The aim of this study was to evaluate the effects of aging on semen quality in a population of infertile couples. Methods: A cross-sectional study of semen samples obtained from 1,500 men randomly selected from couples who attended an infertility clinic was conducted. The analyses were performed using Spearman’s correlation and Mann-Whitney tests. The age groups consisted of men ≤ 35 years, from 36–45 years and > 45 years of age. The semen analysis was performed according to the WHO criteria, and morphology was evaluated using the motile sperm organelle morphology examination (MSOME). The percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV, occupying > 50% nuclear area) were determined. The percentages of DNA fragmentation were assessed using the TUNEL assay. Results: A regression analysis revealed that the percentages of LNV spermatozoa and sperm DNA fragmentation positively correlated with age. Conversely, a regression analysis revealed that the percentage of normal sperm, sperm progressive motility and sperm vitality negatively correlated with age. As in the previous test, the analysis by age group showed that there was a significant reduction (P < 0.05) in the percentage of normal sperm, sperm progressive motility and sperm vitality as age increased. Conversely, the percentage of spermatozoa with LNVs and sperm DNA fragmentation significantly increased (P < 0.05) as age increased. Conclusion: Semen quality seems to be influenced by aging. The age-related decrease in sperm quality suggests that delaying childbearing, not only for women but also for men, may jeopardize reproductive capacity.

Social oocyte cryopreservation: a portrayal of Brazilian women

Objective This study aimed to determine what Brazilian childless women of reproductive age think about oocyte cryopreservation to postpone pregnancy and their reasons for performing or not performing this procedure. Methods Women of reproductive age were randomly selected from the general population using different e-mail lists and were invited to participate in the study by completing an online web survey regarding social oocyte cryopreservation. The survey was also distributed through social media to women of reproductive age. Results Although most of the responders had a partner (86.9%) and had already planned the pregnancy of their first child (69.6%), 85.4% (379) considered the potential of social oocyte freezing to improve their chances of giving birth later in life. Those that had already planned pregnancy were two times more likely to intend to freeze their oocytes (p=0.03). The most important barrier for not undergoing oocyte cryopreservation was cost. The women who indicated that they could not currently undergo the procedure now because of cost were two times (p=0.03) more likely to intend to cryopreserve their oocytes than women who thought that they would not need to delay pregnancy. Conclusion Brazilian women who think that they are not ready to have a family are discovering the option of oocyte cryopreservation. Most participants considered safeguarding their reproductive potential. Making the procedure more accessible could give women the opportunity to make proactive decisions about the future of their fertility.