D. Loutradis

Oocyte morphology correlates with embryo quality and pregnancy rate after intracytoplasmic sperm injection

OBJECTIVEnTo evaluate the relation of oocyte morphology with embryo quality and pregnancy rates (PRs) after intracytoplasmic sperm injection (ICSI).nnnDESIGNnRetrospective study of patients undergoing ICSI.nnnSETTINGnUniversity Hospital IVF Center.nnnPATIENT(S)nSixty-eight patients who underwent ICSI and had transfer of good-quality embryos (grade 3), 60 patients with transfer of both good- and poor-quality embryos (grade 3 and grade 2), and 18 patients with transfer of poor-quality embryos (grade 2).nnnINTERVENTION(S)nComparison of the outcome of ICSI in the three groups of patients and the relation of oocyte morphology to embryo quality.nnnMAIN OUTCOME MEASURE(S)nOocyte morphology and embryo quality (grade). Fertilization, cleavage, and pregnancy rates. Serum E2 on the day of hCG administration.nnnRESULT(S)nOocytes with poor morphology (dark cytoplasm; many vacuoles or fragments in cytoplasm) led to poor-quality embryos and consequently to lower PRs (5.5% versus 29.4%). Serum E2 on the day of hCG administration was significantly higher in the group with good-quality embryos compared with that with poor-quality embryos (2,047 +/- 135.7 versus 1,651 +/- 164.8 pg/mL, respectively).nnnCONCLUSION(S)nSerum E2 on the day of hCG administration is a marker of embryo quality. Oocyte morphology correlates well with embryo quality and PRs after ICSI.

FSH receptor gene polymorphisms have a role for different ovarian response to stimulation in patients entering IVF/ICSI-ET programs

Purpose: To examine the frequency distribution of the Ser680Asn polymorphism of the follicle-stimulating hormone receptor (FSHR) gene in ovarian dysfunction (OD) infertile women, “poor responders” (PR) and “good responders” (GR).n Methods: The hormonal profiles and treatment of all patients were analyzed and FSHR polymorphism was examined by PCR and RFLP. Women from all groups were classified as Asn/Asn, Asn/Ser, and Ser/Ser genotypes.n Results: The frequency distribution of Ser/Ser, Asn/Ser and Asn/Asn variants in OD patients was 45.5, 22.7, and 31.8%, respectively. Day 3 FSH levels in OD and GR patients were higher in Ser/Ser and Asn/Asn subgroups. Asn/Ser carriers from OD and GR groups provided more follicles and oocytes compared to other allelic variants.n Conclusions: GR patients carry more often the Asn/Ser genotype. The latter is correlated with more follicles and oocytes in both OD and GR patients. The Ser/Ser variant might be related to higher serum FSH levels, while the Asn/Ser with lower.

Factors controlling blastocyst implantation

Establishment of early pregnancy is promoted by a complex network of signalling molecules that mediate cell-to-cell and cell-to-extracellular matrix communications, in order to manifest controlled invasion of the trophectoderm and successful implantation. During the period known as the window of implantation, the endometrium expresses specialized proteins, many of which exhibit potential use as markers of endometrial receptivity. Trophoblast differentiation to the invasive phenotype also depends on the up-regulation of certain peptides and the down-regulation of others. Disruption of each pathway is theoretically possible, and studies in animal models suggest that implantation defects result when the function of these proteins is blocked. Indeed, the implantation process is currently considered the most important limiting factor for the establishment of viable pregnancy. The present overview of the literature reports critical families of molecules located at the embryo-maternal interface and describes the mechanisms of interplay and control. Since these factors are crucial to the process of implantation, targeting them might be a valuable contraceptive tool. Conversely, induction of specific peptides may prove to be beneficial in certain infertility cases.

Birth of two infants who were seronegative for human immunodeficiency virus type 1 (HIV-1) after intracytoplasmic injection of sperm from HIV-1-seropositive men

OBJECTIVEnTo report two cases of live births after intracytoplasmic sperm injection (ICSI) in two women who were seronegative for human immunodeficiency virus type 1 (HIV-1) after the use of processed semen from their seropositive husbands.nnnDESIGNnCase reports.nnnSETTINGnUniversity hospital IVF center.nnnPATIENT(S)nTwo HIV-1 seropositive men and their HIV-1 seronegative female partners; all gave their informed consent in writing before undergoing the ICSI procedures.nnnINTERVENTION(S)nThe men provided semen samples that were processed with the use of Percoll and swim-up techniques. Ovarian stimulation in the women was performed with the long protocol using GnRH analogs and recombinant FSH. ICSI was performed.nnnMAIN OUTCOME MEASURE(S)nOocytes were fertilized by ICSI, and the resulting embryos were transferred to the patients. The mothers and babies were tested for HIV-1 antibodies.nnnRESULT(S)nIn the first case, seven mature oocytes were collected and fertilized with ICSI, and three embryos were transferred; the woman became pregnant and gave birth to a healthy boy. Six months after the birth, testing for HIV-1 antibodies in the woman and the baby gave negative results. In the second case, 10 mature oocytes were collected and fertilized with ICSI, and four embryos were transferred; the second woman became pregnant and also gave birth to a healthy boy. Testing for HIV-1 antibodies at the babys delivery also gave negative results.nnnCONCLUSION(S)nIn women who are infertile because of fallopian tube obstruction or in men who have poor quality semen for artificial insemination, ICSI can be performed using processed semen. This method, which involves the use of only one spermatozoon per oocyte, provides HIV-1 seropositive men with the opportunity to have children with a minimal risk-if any-of infecting their female partners.

Biological Factors in Culture Media Affecting in Vitro Fertilization, Preimplantation Embryo Development, and Implantation

Abstract: Optimal culture conditions are of paramount importance for in vitro fertilization of gametes, preimplantation embryo development, and implantation for all species. Water is the basis of all culture media, and ultrapure water should be employed. The main energy sources of a medium are lactate, pyruvate, and glucose. The concentrations of the first two vary in different media, whereas the latter is necessary mainly for the later stages (morula to blastocyst) of development. A fixed nitrogen source is essential for implantation embryo development whether this is provided by amino acids, albumin, or serum. Suboptimal culture conditions can block development. Pronuclear zygotes of most species (but not human) arrest at some point between the two‐cell and the 16‐cell stage. Modifying culture conditions can lead the embryos to develop through this block. Hypoxanthine also causes a two‐cell block to mouse pronuclear zygotes, and this again depends largely on culture conditions. Simple culture media are bicarbonate‐buffered systems with pyruvate, lactate, and glucose. Complex media, such as Hams F‐10, contain in addition amino acids and other elements found in serum. Human tubal fluid simulates the fallopian tube microenvironment. EDTA, gonadotropins, growth factors, and other substances can be included in the media to stimulate development. Coculture of embryos with oviductal cells has shown promising results.

Different ovarian stimulation protocols for women with diminished ovarian reserve.

PurposeTo review the available treatments for women with significantly diminished ovarian reserve and assess the efficacy of different ovarian stimulation protocols.MethodsLiterature research performed among studies that have been published in the Pubmed, in the Scopus Search Machine and in Cohrane database of systematic reviews.ResultsA lack of clear, uniform definition of the poor responders and a lack of large-scale randomized studies make data interpretation very difficult for precise conclusions. Optimistic data have been presented by the use of high doses of gonadotropins, flare up Gn RH-a protocol (standard or microdose), stop protocols, luteal onset of Gn RH-a and the short protocol. Natural cycle or a modified natural cycle seems to be an appropriate strategy. Low dose hCG in the first days of ovarian stimulation has promising results. Molecular biology tools (mutations, single nucleotide polymorphisms (SNPs)) have been also considered to assist the management of this group of patients.ConclusionsThe ideal stimulation for these patients with diminished ovarian reserve remains a great challenge for the clinician, within the limits of our pharmaceutical quiver.

Alternative Approaches in the Management of Poor Response in Controlled Ovarian Hyperstimulation (COH)

Abstract: Improving pregnancy rates in patients with many failed attempts remains a challenge during IVF‐ET or ICSI‐ET programs. The availability of good‐quality oocytes is a prerequisite for good results in such programs. The use of a personalized protocol for controlled ovarian hyperstimulation (COH) that gives the best possible results for the specific patient is a main factor for the success in IVF or ICSI. The response of many patients to the ovarian stimulation used is very poor, giving fewer oocytes than expected, resulting in much lower, if any, pregnancy rates. The definition of a poor responder is not clear and differs among researchers. A variety of strategies have been used to improve response in these patients, regardless of the definition used. These include various ovulation induction protocols that we believe might assist these patients achieve a pregnancy. The difficulty is greater due to the fact that poor responders are not a homogeneous group and each patient may have a different cause. More studies with large numbers of patients are needed in order to find those protocols that could provide these couples with an acceptable pregnancy rate.

The Validity of Gynecological Ultrasonography

The present study was undertaken to examine the reliability of the sonographic diagnosis in 705 gynecological patients. The determination of the lesions was defined according to the operative diagnosis. The sensitivity, specificity and positive predictive value of the ultrasound technique were evaluated using the surgical findings as gold standard. In 631 patients (89.5%) the ultrasound examination established a correct diagnosis. The sensitivity and specificity of the ultrasound examination varied between 75-95.3% and 93.3-100%, respectively. The positive predictive value was found between 89.7 and 100%, while the false sonographic results were 10.4%, which included those of ectopic pregnancies. It is thus concluded that ultrasonography as compared to the surgical findings has proved to be of great value in establishing a gynecological diagnosis.

Oocyte donation to women over 40 years of age: pregnancy complications

Recently, oocyte donation to women of advanced age has led to a considerable number of conceptions, thus increasing the age limit for becoming pregnant. A main consideration encountered by physicians, though, is the potential medical and obstetric complications of a pregnancy at an advanced age. In this study, the obstetric complications, as well as the perinatal outcome, of pregnancies of aged recipients (above 40) are presented and compared to those of younger recipients. A significantly higher incidence of gestational diabetes (P < 0.001), an increased incidence of pre-eclampsia (at the 10% level of significance) and an increased risk for thrombophlebitis (again at the 10% level) was observed in the older patients, but a careful follow-up during their pregnancy led to a highly satisfactory obstetric and perinatal outcome. A rigorous precycle medical screening (especially for cardiovascular diseases and diabetes) and a careful follow-up during pregnancy is, therefore, imperative so that oocyte donation to older women is not withheld and continues to provide fertility possibilities to otherwise sterile patients.

A preliminary study of the effect of growth hormone on mouse preimplantation embryo development in vitro

The role of growth hormone (GH) in follicular development, ovulation and embryo development is currently under reconsideration. In this study, we have tried to investigate the effect of GH on preimplantation development of mouse embryos in vitro. Zygotes and two-cell mouse embryos were cultured without (control) or with GH. For zygotes, the addition of 0.2 micrograms/ml of GH resulted in 77 +/- 1% of blastocysts formed and 66 +/- 3% rate of hatching (control 64 +/- 4 and 31 +/- 3%, p < 0.05 and p < 0.01, respectively). For two-cell embryos, the addition of 0.2 micrograms/ml of GH resulted in 87 +/- 2% of blastocysts formed and 60 +/- 4% hatching rate (control 76 +/- 4 and 47 +/- 5%, p < 0.05 for both). This positive effect of GH addition implies that the latter can support mouse preimplantation development in vitro and it suggests, along with its local action on the ovary and its possible effects, via the insulin-like growth factor system, on the tubal and uterine epithelium, a continuous role of this hormone in reproductive physiology from follicular maturation to embryonic development and, possibly, implantation.