Vincent W. Aoki

Body mass index is inversely related to intra-follicular HCG concentrations, embryo quality and IVF outcome

Decreased periovulatory human chorionic gonadotrophin (HCG) concentrations have been shown to be associated with diminished fertilization rates. This study evaluated if intra-follicular HCG concentration may be related to body mass in 247 IVF patients using their own oocytes and 58 patients receiving donor oocytes, and evaluated if such a relationship might affect IVF outcome. A significant inverse correlation (r = -0.353, P < 0.001) was observed between the body mass index (BMI) and intra-follicular HCG concentration. The mean HCG concentrations were significantly decreased (P < 0.001) in patients with a BMI >30 kg/m(2) compared with patients with a BMI of 20-30 kg/m(2) or BMI <20 kg/m(2) (17.6 versus 45.1 and 52.5%, respectively). The clinical pregnancy rates (P < 0.001) and embryo quality (P < 0.05) were significantly different for the three groups. In donor oocyte recipients, the pregnancy rate was significantly decreased (P < 0.0001) for recipients with a BMI >25 kg/m(2) compared with those with a BMI from 21-25 kg/m(2) and BMI <21 kg/m(2) (43.8 versus 72 and 76.5%, respectively). These data indicate that intra-follicular HCG concentration is inversely related to BMI, and may be related to a concurrent decrease in embryo quality and pregnancy rates.

Comparison of four media types during 3-day human IVF embryo culture

The aim of this study was to compare the effectiveness of human tubal fluid (HTF), G1.2, Sage Cleavage and Life Global media for IVF outcome during 3-day culture of human embryos. A three-phase auto-controlled study was conducted in which IVF outcome was compared between (1) HTF and G1.2, (2) HTF and Cleavage, and (3) Cleavage and Life Global. In phase 1, no differences in embryo quality were observed between HTF and G1.2. However, embryos derived from intracytoplasmic sperm injection (ICSI) displayed significantly improved quality when grown in HTF versus G1.2. No differences in pregnancy and implantation rates were observed in cases where embryos transferred were grown exclusively in HTF or G1.2 media. In phase 2, embryo quality was significantly improved for embryos cultured in Cleavage versus HTF media (P < 0.001). However, pregnancy, implantation and spontaneous abortion rates were similar between the two media. In phase 3, there were no differences in embryo quality, pregnancy, implantation, and spontaneous abortion rates between Cleavage and Life Global media. Overall, the data indicate that Life Global and Cleavage media yield similar results in a 3-day IVF culture programme. Cleavage medium is superior to HTF, as evidenced by significantly improved embryo quality (P < 0.001). Meanwhile, HTF medium is superior to G1.2 for ICSI cases.

Identification of Genetic Variation in the 5′ and 3′ Non-coding Regions of the Protamine Genes in Patients with Protamine Deregulation

Deregulation of sperm nuclear protamine ratio (P1/P2) has been shown to correlate with male factor infertility in humans, but the cause of this abnormal protein expression has yet to be identified. Recent studies have shown that there is little genetic variability in the coding regions of either of the protamine gene sequences. However, these studies did not investigate the 5′ or 3′ non-coding regions of these genes for mutations that might account for changes in the transcriptional or translational regulation of the protamines. In an effort to determine if genetic variation in these non-coding regions may account for aberrant protamine expression, we have sequenced the 5′ and 3′ untranslated regions (UTRs) of both protamine 1 (P1) and protamine 2 (P2) genes in a population of infertile men with protamine deregulation, men presenting for infertility work-up with normal protamine ratios, and a population of unrelated, fertile men from the Utah Genetic Reference Project (UGRP). This analysis has identified 14 single nucleotide polymorphisms (SNPs), of which 13 were novel SNPs in the UTRs of P1 and P2, and verified the existence of a variable length repeat (VLR), GAn, in the P2 5′ region. The SNP frequencies and VLR allelic frequencies did not achieve statistical significance between the populations, however, one of the SNPs identified in the 3′ UTR of protamine 2 was found at a low frequency in the abnormal protamine patients, but was completely absent in men with verified normal protamine ratio and donors of known fertility. In conclusion, a number of SNPs have been reported in the protamine genes and the untranslated regions, however, these gene variants do not appear to be responsible for protamine deficiency. Hence, the underlying cause for aberrant protamine expression may possibly be due to abnormalities in candidate spermatogenic transcriptional/translational regulators, post-translational modifiers, or as-of-yet unidentified factors affecting the testicular environment.

Correlation of sperm penetration assay score with polyspermy rate in in-vitro fertilization

Background The sperm penetration assay (SPA) is used to predict the fertilizing capacity of sperm. Thus, some programs rely on SPA scores to formulate insemination plans in conjunction with in-vitro fertilization (IVF) cycles. The purpose of this study was to evaluate if a relationship exists between SPA scores and polyspermy rates during conventional IVF cycles. Methods A total of 1350 consecutive IVF patients using conventional IVF insemination were evaluated in the study. Oocytes were inseminated three hours post-retrieval by the addition of 150,000 to 300,000 progressively motile sperm. Approximately 18 hours after insemination, the oocytes were evaluated for fertilization by the visualization of pronuclei. The presence of three or more pronuclei was indicative of polyspermy. Polyspermy rates, fertilization success, embryo quality, and pregnancy rates were analyzed retrospectively to evaluate their relationship with SPA score, count, motility, number of progressively motile sperm inseminated, oocyte pre-insemination incubation time, patient age, and diagnosis. Results A significant positive relationship was observed between SPA score and polyspermy rate (rs = 0.10, p < 0.05). Patients with a normal SPA score had significantly higher polyspermy rates than those with abnormal SPA scores (6.3% ± 1.5% vs. 2.0% ± 0.7%, p < 0.05). Fertilization percentage was significantly lower in the group with severely abnormal SPA scores versus all other SPA groups (57.5% ± 2.1% vs. 70.2% ± 1.3%, p < 0.005). Although embryo quality was not affected, both clinical pregnancy and implantation rates improved slightly as SPA score increased. In addition, there was a decrease in the rate of spontaneous abortion as SPA score increased. Conclusions These data indicate SPA score is positively correlated with polyspermy rates and IVF fertilization percentage. Additionally, there is a slight increase in clinical pregnancy rates, and embryo implantation rates with increased SPA. Furthermore, there is a slight decrease in spontaneous abortions rates related to increased SPA.

Evaluation of the developmental competence and chromosomal compliment of mouse oocytes derived from in-vitro growth and maturation of preantral follicles

PurposeTo evaluate the developmental potential and aneuploidy rates of in-vitro versus in-vivo grown and matured mouse oocytes.MethodsMice were superovulated to obtain in-vivo matured oocytes. Mouse preantral follicles were also mechanically isolated and cultured in-vitro. In-vitro fertilization (IVF) was performed and fertilization, cleavage, and morula/blastocyst formation rates were compared between groups. Cytogenetic analysis was used to compare oocyte aneuploidy rates and aneuploidy characteristics in the developing embryos.ResultsIn-vivo oocyte maturation resulted in higher IVF fertilization, cleavage, and morula/blastocyst formation rates versus in-vitro follicle culture (96.4% versus 78.5%, p < 0.001; 95.3% versus 77.4%, p < 0.001; 94.1% versus 76.9%, p < 0.001). Total aneuploidy rates were higher in embryos derived from in-vitro matured oocytes versus those grown in-vivo (4.0% versus 1.3%, p < 0.05).ConclusionsResults indicate a reduced developmental competency of in-vitro matured oocytes. The data also highlight an increased susceptibility to meiotic errors in early stage follicles undergoing in vitro culture.

Collaboration Within the Reproductive Endocrinology and Infertility Practice: Integration of Laboratory and Clinic Operations

The quality of patient care in the reproductive medicine clinic hinges on effective integration of laboratory and clinic operations. The reproductive endocrinology and infertility (REI) practice represents a truly unique niche, intimately partnering the laboratory and clinic in overall patient treatment and management paradigms. As such, to deliver optimal patient care, there are a number of unique challenges an REI clinic faces in effectively integrating the two arenas. This chapter discusses these challenges and outlines methods by which REI practitioners may better integrate clinic and laboratory operations to deliver high quality care to patients. Specific areas of focus include patient care and management, quality management programs, regulatory affairs, risk management, continuing education, and professional development.

The Genetics of Abnormal Protamine Expression

During spermiogenesis, the sperm chromatin undergoes dramatic remodeling. The testis-specific protamine proteins facilitate these nuclear changes by replacing the somatic cell histones, a process that produces highly condensed, transcriptionally silent chromatin. In humans, there are two forms of sperm protamine: protamine 1 (P1) and protamine 2 (P2), which occur in a strictly regulated 1:1 ratio. Sperm protamine-deficiency and P1:P2 ratio deregulation have been implicated in male infertility. The details of the underlying genetic basis of abnormal protamine expression are just emerging. This chapter summarizes our current knowledge of the sperm protamines, their relationship with male infertility, and what is currently understood regarding the genetic basis of abnormal protamine expression.