Silvina Bocca

Fragmentation of DNA in morphologically normal human spermatozoa

OBJECTIVE To evaluate DNA fragmentation in spermatozoa with normal morphological appearance. DESIGN Prospective study. SETTING Academic tertiary center. PATIENT(S) Fertile, subfertile, and infertile men were studied. INTERVENTION(S) Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-fluorescein nick-end labeling assay and morphology assessment by phase contrast in the swim-up fractions. MAIN OUTCOME MEASURE(S) Simultaneous assessment of the percentage of normally shaped sperm and DNA fragmentation. RESULT(S) No DNA fragmentation was found in spermatozoa with normal morphology in any of the samples from the fertile group. In only one sample from the subfertile group did we observed normally shaped sperm cells exhibiting DNA fragmentation. However, in all the samples from the infertile group, we observed normal spermatozoa with DNA fragmentation. Spermatozoa from this late group exhibited a high proportion of DNA damage. CONCLUSION(S) In infertile men with moderate and severe teratozoospermia, the spermatozoa with apparently normal morphology present in the motile fractions after swim-up may have DNA fragmentation.

The Z Technique: An Easy Approach to the Display of the Midcoronal Plane of the Uterus in Volume Sonography

Objective. The purpose of our study was to evaluate the ease of learning and clinical applicability of a technique (Z technique) for display of the midcoronal plane of the uterus in volume sonography. Methods. Eight physicians were randomly assigned to 2 groups, A and B, after being instructed on the basic principles of post processing of sonographic volumes. Physicians in group A were asked to individually display the midcoronal plane of the uterus in each of five 3‐dimensional volumes of uteri. Physicians in group B were instructed on the Z technique and were then asked to individually display the midcoronal plane of the uterus in the same volumes as group A. The time needed for display of the midcoronal plane of the uterus and the quality of the midcoronal plane image retrieved were recorded for each volume. Results. The mean time required to display the midcoronal plane of the uterus for all volumes was 47.7 seconds in group B compared with 110.7 seconds in group A (P = .002). Furthermore, a significant improvement in time performance was seen for physicians in group A after learning the Z technique (23.2 seconds after compared with 110.7 seconds before; P = .0001). The quality of the image produced was notably better for all physicians after learning the Z technique. Conclusions. The Z technique is an easy technique to learn. Physicians who learn this technique are able to retrieve the midcoronal plane of the uterus faster and improve its image quality in volume sonography.

Anti-Müllerian hormone serum levels predict response to controlled ovarian hyperstimulation but not embryo quality or pregnancy outcome in oocyte donation

The objective of this retrospective cross-sectional study was to evaluate the value of basal serum anti-Müllerian hormone (AMH) levels as a predictor of ovarian response and pregnancy outcome in a donor egg program. The study showed that AMH was superior to other biomarkers of ovarian reserve in predicting low and high response in young women selected as oocyte donors, but that it was not predictive of embryo morphology or pregnancy outcome in the recipient population.

A cross-sectional evaluation of the first cohort of young adults conceived by in vitro fertilization in the United States

OBJECTIVE To assess the quality of life and susceptibility for chronic disease development of the oldest generation of young adults conceived by IVF in the U.S. DESIGN Cross-sectional. SETTING Single tertiary clinic. PATIENT(S) Young adults conceived by standard IVF between 1981 and 1990. INTERVENTION(S) Self-administered questionnaire. MAIN OUTCOME MEASURE(S) Indicators of physical, psychologic, and behavioral health. RESULT(S) A total of 173 (31%) of 560 eligible young adults completed the questionnaire. Mean age was 21.2 years (range 18-26 years) and male-to-female ratio was 3:4. A limited number were conceived through gamete donation but none through oocyte/embryo micromanipulation. Prevalence rates of overweight and obesity were 35% and 10%, respectively. More than 65% were ever diagnosed with a chronic condition; most diagnoses were psychiatric, ocular, respiratory, and cardiometabolic in nature. Almost 40% of respondents were lifetime smokers, 62% reported binge drinking in the previous year, and >90% were physically active in the preceding month. Survey participants were mostly similar to a subsample of the 1999-2004 National Health and Nutrition Examination Survey on selected health indicators. CONCLUSION(S) Young adults conceived by IVF appear to be healthy and well adjusted, although the preponderance of psychologic health problems requires further investigation.

Automated assessment of ovarian follicles using a novel three-dimensional ultrasound software

OBJECTIVE To evaluate the accuracy, efficiency, and applicability of a new automated method of follicular assessment. DESIGN Retrospective review of three-dimensional ultrasonographic ovarian volumes. SETTING Academic center. PATIENT(S) Three-dimensional ovarian volumes from patients undergoing IVF. INTERVENTION(S) Three-dimensional volumes of gonadotropin-stimulated ovaries and simulated ovarian follicles (SOFs) of known volume were evaluated with the new automated follicular assessment software (SonoAVC). MAIN OUTCOME MEASURE(S) [1] Maximum absolute error for the SonoAVC in assessing the volumes of the SOFs; [2] correlations between the automated and manual follicular measurements; [3] time required to analyze all of the follicles in a given ovarian volume. RESULT(S) The SOF of 4, 6, and 10 mm were evaluated. The SonoAVCs maximum absolute error for the volumes of the 4, 6, and 10 mm SOFs was 0 (0%), 0.01 (8.3%), and 0.02 (3.8%) cc, respectively. Three hundred forty-seven follicles, ranging in diameter from 2.3-32 mm, were evaluated. The correlation coefficient for the SonoAVC-generated relaxed follicular diameter and the manual measured mean follicular diameter was 0.99. The time required to analyze all of the follicles in a given ovarian volume for the automated and manual method was 133 versus 361 seconds, respectively. CONCLUSION(S) The SonoAVC proved to be a very accurate and efficient way to measure ovarian follicles. The measurements obtained by the SonoAVC correlated extremely well with the manual measurements we obtained.

Expression of immunomodulatory genes, their protein products and specific ligands/receptors during the window of implantation in the human endometrium

We have demonstrated up-regulation of the immunomodulatory genes decay accelerating factor (DAF), interleukin 15 (IL-15) and osteopontin (OPN) during the window of implantation (WOI). Here, we characterized gene expression and determined the localization of their protein products and respective ligands at the opening and closure of the WOI. In addition, we used laser capture microdissection (LCM) to analyze the cell type-specific gene expression. Human endometrial biopsies from cycle Days 16, 21 and 24 were evaluated by real-time RT-PCR. Purified epithelial and stromal cells were obtained by LCM. Localization of the proteins and their ligands was assessed by immunohistochemistry. mRNA expression of DAF, IL-15 and OPN was significantly increased throughout the WOI. DAF, OPN and alpha(v)beta(3) integrin were strongly immunolocalized to the glandular compartment by Days 21 and 24, whereas C3, IL-15 and IL-15Ralpha were highly stained in both glandular and stromal compartments. After LCM, gene expression of DAF was 4.8-fold increased in epithelium versus stroma, whereas for OPN there was a 2-fold increase. For IL-15, the expression in stroma was 8.7-fold higher than in epithelial cells. The progressive increase of the expression of these immunomodulatory genes, proteins and ligands during the WOI, support a critical role at the time of endometrial receptiveness.

A novel model of human implantation: 3D endometrium-like culture system to study attachment of human trophoblast (Jar) cell spheroids

There is an urgent need to develop optimized experimental models to examine human implantation. These studies aimed to (i) establish a human endometrium-like three-dimensional (3D) culture system, and (ii) examine the attachment of trophoblast-like Jar spheroids to the culture. In the present work, 3D endometrial cultures were constructed with fibrin-agarose as matrix scaffold, and using epithelial and stromal cells from both human primary cultures and established cell lines. An attachment assay between trophoblast cells and the 3D culture was developed. Epithelial cells (cytokeratin(+)) concentrated on top of the matrix forming a monolayer, and stromal cells (vimentin(+)) resided within the matrix, resembling the normal endometrial structure. The capability of primary epithelial cells to form glands spontaneously was observed. Human trophoblast cells (Jar cells) were hCG(+) by immunostaining, allowed to form spheroids, and confirmed to secrete hCG into the medium. Time-dependent experiments demonstrated a high rate of attachment of Jar spheroids to the epithelium, and adhesion was strongly related to the various cell types present in the 3D culture. An architecturally and functionally competent 3D endometrial culture system was established, that coupled with Jar spheroids mimicking trophoblast cells, provides a unique in vitro model for the study of certain aspects of human implantation.

Uncomplicated full term pregnancy after da Vinci-assisted laparoscopic myomectomy

Laparoscopic surgery with the assistance of the da Vinci robotic system has been recently introduced in gynaecology, as this new technology provides three-dimensional vision and easier suture capability. This study reports, for the first time, the case of an uncomplicated full term pregnancy after laparoscopic myomectomy with the assistance of the da Vinci robotic system. The patient was a 35-year-old woman presenting with secondary infertility and a single 3 cm, predominantly intramural, fundal myoma. Following surgery, she conceived with minimal intervention therapy and delivered a healthy term infant by Caesarean section. The successful outcome suggests that this advancement can restore reproductive capacity with improved suturing capability of the uterine wall. More studies are needed to establish indications and outcome of the technique according to the number, size and location of fibroids.

A study of the cost, accuracy, and benefits of 3-dimensional sonography compared with hysterosalpingography in women with uterine abnormalities

We conducted a prospective blinded study to evaluate the costs, accuracy, risks, and benefits of 3‐dimensional (3D) transvaginal sonography compared to hysterosalpingography.

Cryopreserved/thawed semen for in vitro fertilization: results from fertile donors and infertile patients *

We evaluated the in vitro fertilization (IVF) outcome in 54cycles using cryopreserved/thawed semen from fertile donors. Controls were other IVF patients matched by time frame, female age, stimulation protocol, number of pre-embryos transferred, and absence of a male factor using freshly ejaculated normal semen samples. In the study group and controls, respectively, post-thaw swim-up motility was 83.1% and 89.5%; fertilization rate of preovulatory oocytes (91.8%, 95.7%) and ongoing pregnancy rate (PR) per transfer (21.1%, 25.0%) were similar. The excellent fertilization rate with frozen/thawed semen was achieved through high-concentration insemination (0.5 X 10 6 motile sperm/mL). With use of frozen/thawed samples from infertile men (normal and subfertile samples), PR was similar but fertilization rate was lower. Cryopreserved semen is a valuable option for infertile couples in IVF therapy.