To evaluate the efficiency of single and multiple embryo transfers in a donor-recipient program using vitrified/warmed oocytes. Retrospective study. A total of 5014 MII oocytes were obtained and vitrified from 225 donors who were screened and tested according to FDA and ASRM guidelines. Eggs were vitrified 3-4 h after retrieval using minimal volume method. ICSI was performed 2-3 h after warming. Embryos were evaluated morphologically prior to transfer, and when high quality embryos were observed the recipient had the possibility of a single embryo transfer. Clinical and laboratory parameters were analyzed using the One-way ANOVA and the Fisher's exact test at the level of P<0.05. Donors mean age was 26.1 ± 2.7 years.Tabled 1Single-ETMulti-ETPNo. of recipients126351Recipient age (mean ± SD)42.2 ± 4.240.7 ± 4.4<0.01No. of oocytes warmed (mean ± SD)711 (5.6 ± 1.8)2238 (6.4 ± 2.0)<0.01No. of oocytes survived (%)643 (90.4)1957 (87.4)0.04No. of oocytes fertilized (%)560 (78.8)1707 (76.3)NSNo. of good quality embryos on Day3 (mean ± SD)380 (3.0 ± 1.6)1040 (3.0 ± 1.4)NSNo. of blastocysts (mean ± SD)*Only for Day-5 ET cases (113 for SET and 274 for MET).396 (3.5 ± 1.5)976 (3.6 ± 1.5)NSNo of embryos for ET (mean ± SD)126725NANo. of embryos re-vitrified (mean ± SD)265 (2.1 ± 1.5)356 (1.0 ± 1.3)<0.05No. clinical pregnancies (%)77 (61.1)211 (60.1)NSNo. of implantations (%)78 (61.9)317 (43.7)<0.05There were 105 (49.8%) pregnancies with multiple implantations in the MET group, while there was only one twin pregnancy (due to monozygotic twinning) in the SET group (P<0.001).* Only for Day-5 ET cases (113 for SET and 274 for MET). Open table in a new tab There were 105 (49.8%) pregnancies with multiple implantations in the MET group, while there was only one twin pregnancy (due to monozygotic twinning) in the SET group (P<0.001). The outcome of the current study shows that SET of vitrified/warmed donor oocytes provides both high implantation rates (and virtually no risks for multiples) and high pregnancy rates similar to multiple embryo transfers. Thus, SET should be recommended when high quality embryo(s) are available for qualifying patients to achieve safe pregnancy outcomes.

Using sibling oocytes, the objective of this study was to compare the intracytoplasmic sperm injection (ICSI) fertilization rates to those achieved with conventional IVF in patients with high rates of oocyte immaturity. This study was observational in nature, and included 91 patients who were treated using split insemination techniques. The fertilization rates for the ICSI group and the IVF group were 41.1 +/- 15.0% and 53.2 +/- 19.8%, respectively (P <: 0.0001). There was no significant difference in day-3 embryo quality between the two groups. There was a significantly higher number of embryos frozen in the IVF group than in the ICSI group: 357 (84.8%) and 297 (76.7%), respectively (P = 0.037). Furthermore, the number of embryos either transferred or frozen was significantly higher in the IVF group than the ICSI group: 459 of 1173 (39.1%) and 385 of 1268 (30.4%), respectively (P < 0.0001). These data indicate that conventional IVF results in a higher fertilization rate than ICSI. Furthermore, IVF provided more embryos available for transfer or cryopreservation when compared with ICSI, thereby optimizing the patient's cycle.

Objective: To determine the relationship between platelet-activating factor acetylhydrolase (PAFah) content in semen and sperm motility.Design: The PAFah levels in semen were measured and correlated with sperm motility.Setting: Clinical laboratory in a private assistant reproductive technology clinic.Patient(s): Three hundred and twelve men seeking diagnosis and treatment of infertility.Intervention(s): Semen samples were collected from 312 healthy mature men seeking infertility treatment. Sperm motility and PAFah activity were measured in seminal plasma. Data was analyzed by Student's t test and regression analysis.Main Outcome Measure(s): PAFah activity and sperm motility.Result(s): Seminal PAFah content ranged from a low of 179 IU/L to a high of 27457 IU/L. The overall mean PAFah content in semen was 780.59 IU/L. Linear regression analysis revealed a significant (R-2 = 0.655) and negative relationship between PAFah content in semen and sperm motility. Semen specimens with high percent motility (>= 50%) had significantly lower PAFah concentrations (442.03 +/- 14.37 IU/L) than those with the lower percent sperm motility (< 50%) (882.16 +/- 18.45 IU/L).Conclusion(s): The data confirm the presence of PAFah in human semen and that activity is significantly and negatively correlated with sperm motility. The PAFah is proven to be a candidate for sperm decapacitation factors, whereas PAF is qualified to be a candidate for sperm capacitation factors.

The objective of the present study was to compare a traditionally used bovine-derived hyaluronidase (Hyase) with the newly developed recombinant human-derived enzyme product (Cumulase) in intracytoplasmic sperm injection (ICSI) procedures using a sibling oocyte model in a prospective randomized design. The results of the Study demonstrate that Cumulase is safe and effective in an ICSI treatment program and can provide comparable if not improved parameters, including fertilization and embryo developmental rates.

Purpose : A major gene responsible for the control of preimplantation cleavage rate is the Ped gene, the product of which is the Qa-2 protein. Fast, but not slow developing mouse embryos express the Qa-2 protein. Platelet-activating factor (PAF) is a novel and potent signaling phospholipid that has unique pleiotropic properties in addition to platelet activation. PAF plays a significant role in virtually every reproductive event, including ovulation, fertilization, implantation, and parturition. The role of the Ped gene in PAF production by preimplantation embryos is yet to be established. The presence of this gene provides embryos with a reproductive advantage over those that are Ped negative, and may also serve as a regulator of PAF synthesis. The study hypothesis is that the amount of PAF produced is dependent upon the presence or absence of the Ped gene.Methods : B6.K1 (Ped negative) and B6.K2 (Ped positive) mouse embryo-conditioned culture media were assayed for PAF content by a PAF-specific radioimmunoassay.Results : There was a significant (p < 0.001) difference in blastocyst development rates between the Ped+ B6.K2 (61.0%) and the Ped- B6.K1 (25.3%) embryo culture groups. There was a significant difference (p < 0.05) in PAF production between the Ped+ B6.K2 (4.70 +/- 0.46 pmol per embryo) embryo culture group and the Ped- B6.K1 (10.02 +/- 3.49 pmol per embryo) embryo group. The B6.K1 (Ped-) embryo group produced > 2x more PAF than did the B6.K2 (Ped+) group.Conclusions : The Ped gene plays a role in PAF production and release in preimplantation stage embryos. The use of two mouse identical strains, except for the Ped gene, show that its presence is associated with an increase in developmental potential. Embryos where the Ped gene was absent produced significantly higher levels of PAF, which may aid in their survival.

OBJECTIVE:To evaluate cycle outcomes in patients with either poor or normal prognosis undergoing IVF treatment with a GnRH antagonist (ganirelix acetate) for LH suppression. DESIGN:Nonrandomized, noncontrolled, retrospective review. PATIENT(S):204 patients, aged 23-41 years, undergoing IVF. INTERVENTION(S):Patients completed 225 consecutive cycles of IVF with a GnRH antagonist (Antagon; Organon, Roseland, NJ) for LH surge prevention. Sixty cycles were conducted in patients with a known poor prognosis, whereas 165 were conducted in patients with a normal IVF prognosis. MAIN OUTCOME MEASURE(S):Pregnancy rate (PR), for the series as a whole and according to prognosis, and serum E2 patterns. RESULT(S):The PR per initiated cycle for the series as a whole was 33.3%. The pregnancy rate was 42.1% per ET for the entire series, with a cycle cancellation rate of 21%. When evaluated by prognosis, the poor-prognosis patients had PRs of 8.3% per attempt and 15% per transfer, whereas the normal-prognosis patients had PRs of 40% per attempt and 45% per transfer. Pregnancy rate did not vary by E2 pattern (drop, plateau, or rise). Oral contraceptive pretreatment was noted to be associated with high cancellation rates in the group of known poor responders, whereas for the group as a whole, cycle outcome was unaffected by the use of oral contraceptives. CONCLUSION(S):Use of GnRH antagonists in patients with an a priori poor IVF prognosis results in predictably poor outcomes. Patients without factors predicting poor outcome have acceptable PRs. The pattern of E2 rise immediately after initiation of GnRH antagonists does not predict cycle outcome. Oral contraceptives can be successfully used to schedule antagonist-based IVF cycles but might increase the risk of cycle cancellation in some patient populations.

Since its discovery nearly 30 years ago platelet-activating factor (PAF) has emerged as one of the more important lipid mediators known. PAF (1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphorylcholine) exists endogenously as a mixture of molecular species with structural variants of the alkyl moiety. PAF is a novel potent signaling phospholipid that has unique pleiotropic biological properties in addition to platelet activation. PAF also plays a significant role in reproduction and is present in the sperm of a number of primate species. PAF content in squirrel monkey sperm is significantly higher during the breeding season than the non-breeding season. PAF content in rhesus sperm has a significant relationship with sperm motility. PAF content in human sperm has a positive correlation with seminal parameters and pregnancy outcomes. The enzymes (lyso-PAF-acetyltransferase and PAF-acetylhydrolase) necessary for PAF activation and deactivation are present in primate sperm. PAF-acetylhydrolase may act as a "decapacitation factor". Removal of this enzyme during capacitation promotes PAF synthesis increasing primate motility and fertilization. PAF also plays a significant role in the fertilization process, enhancing the fertilization rates of oocytes. Enhanced embryo development has also been reported in oocytes fertilized with PAF-treated sperm. Exogenous PAF will also significantly improve primate artificial insemination pregnancy outcomes. PAF antagonists inhibit sperm motility, acrosome reaction, and fertilization thus suggesting the presence of receptors for PAF. The PAF-receptor is present on primate sperm, with altered transcript levels and distribution patterns on abnormal cells. Whereas, the exact mechanism of PAF in sperm function and reproduction is uncertain, its importance in normal primate fertility is substantial.

Body mass index (BMI) has been demonstrated to affect female fertility; however, little information is available on the impact of BMI on male fertility or semen parameters. Therefore, the study objective was to determine the relationship between BMI and semen parameters, including sperm chromatin integrity. We analyzed data on semen samples from 520 men who were grouped based upon calculated BMI values (normal, 20-24 kg/m(2); overweight, 25-30 kg/m(2); obese, >30 kg/m(2)). The data collected included patient height and weight, semen volume, sperm concentration, percent sperm motility, percent sperm morphology (normal forms), and sperm chromatin integrity (DNA fragmentation index [DFI]). Data were analyzed by regression analysis and analysis of variance (ANOVA) with Tukey's test for multiple pairwise comparisons. The overall BMI mean (+/-SEM) was 27.5 (+/-0.49) kg/m(2). Linear regression revealed a significant (P < .05) and negative relationship between BMI and the total number of normal-motile sperm cells. ANOVA revealed a significant difference (P < .05) in the total number of normal-motile sperm cells among the different BMI groups. The number of normal-motile sperm cells per BMI group was as follows: normal, 18.6 x 10(6); overweight, 3.6 x 10(6); and obese, (0.7) x 10(6). All multiple pairwise comparisons were found to be significantly (P < .05) different. The overall DFI mean (+/-SEM) was 24.7 (+/-2.57). Linear regression revealed a significant (P < .05) and positive relation between BMI and DFI. Men presenting with a BMI greater than 25 kg/m(2) have fewer chromatin-intact normal-motile sperm cells per ejaculate. Therefore, to ensure maximum fertility potential, patients may be advised to reduce body weight.

Objective: To investigate quantitative aberrations involving p53 copy numbers in eutopic endometrial and endometriotic tissue from two populations.Design: Comparative analysis of normal and diseased tissue.Setting: Tissue specimens collected in Iceland and USA.Patient(s): Subjects with moderate/severe endometriosis (Iceland, n = 26; USA, n = 45). Paraffin-embedded tissue from 19 matched Icelandic cases and seven unaffected controls. American cases were fresh surgical tissue from 17 matched cases and 28 unaffected controls. DNA isolation and real-time polymerase chain reaction (PCR) with TaqMan assay were performed.main Outcome Measure(s): The frequency of p53 loss and/or gain based on quantitative differences for copy numbers of p53 located on chromosome (17p) and GAPDH on a control locus (chromosome 12p).Result(s): Among American cases, significant p53 gain (n = 13) or loss (n = 4) was observed in 17 of 21 cases. In Icelandic cases this was not seen to the same degree. Mean normalized p53 values were 3.46 and 1.16 copies per reaction, respectively. Significant differences were observed between normalized p53 in the control blood and affected tissue for the American and Icelandic cases compared to standard GAPDH control but not in normal Icelandic and American endometrium.Conclusion(s): The results continue to support a role for nonrandom somatic p53 locus alterations in the pathogenesis of late or sever-stage endometriosis. Differences between Icelandic and American subjects have implications for generalization of genome-wide approaches.

Problem: Platelet-activating factor (PAF) plays a significant role in fertility. Preimplantation stage embryos produce PAF (ePAF) which is required for development. PAF's mechanism of action is receptor-mediated and its presence has been reported in the developing mouse and human embryo. Exposure of preimplantation stage mouse embryos results in higher implantation rates. However, the effect of such treatment on live-birth rates and birth weights has not been reported. Therefore, the objective the study was to determine the effect of exposing preimplantation mouse embryos to PAF on subsequent birth rate and weight.

Introduction: Previous studies have demonstrated the presence of ribonucleic acid (RNA) in mature, ejaculated sperm and that sperm with poor motility have significantly more RNA than good motility sperm. However, no studies have investigated the relationship between RNA content and sperm morphology. Therefore, the study objective was to determine the relationship between total RNA content and sperm morphology. Methods: Sperm samples were separated by density centrifugation (Promotor, CERES Fertility, San Diego, CA) into morphologically normal and abnormal populations. Sperm morphology scores were determined by strict criteria. Total RNA was extracted from sperm via the Roche (Indianapolis, IN, USA) RNA isolation kit. Total RNA (μg of RNA per million sperm cells) recovered per sperm population from each ejaculate was determined by spectrophotometer and calculated as follows: (absorbance at 260 nm)×(conversion factor)×(dilution factor). The conversion factor for RNA is 0.04 μg/μl per optical density unit at 260 nm. Total RNA purity was calculated as the absorbance at 260 nm divided by the absorbance at 280 nm. Data were analyzed by Student's t-test. Results: A total of 179 semen specimens from different men underwent RNA extraction and analysis as described. All samples had absorbance readings between 0.1 and 1.0 at 260 nm and were determined to be accurate. Total RNA purity estimates for all samples were within the acceptable ratio of 1.7 and 1.9. RNA content in morphologically normal sperm (2.83±0.14 μg/106 cells) was significantly higher (P<0.05) than the amount found in morphologically abnormal sperm (2.26±0.23 μg/106 cells). Conclusions: Ribonucleic acid content is significantly lower in morphologically abnormal sperm. Morphologically abnormal sperm may have impaired transcriptional and, or translational pathways thus negatively impacting cell function.

Objective: To determine the efficacy of treating semen specimens with platelet-activating factor (PAF) before IUI.Design: Prospective randomized double-blinded study of PAF treatment of sperm for patients with a history of infertility undergoing IUI.Setting: Private infertility center.Intervention(s): Patients had ovulation induction therapy with clomiphene citrate (CC) or gonadotropin, two IUIs per month with PAF treatment.Main Outcome Measure(s): Clinical pregnancy rates.Result(s): There was a significant difference in IUI pregnancy rates per cycle between control (10/56; 17.9%) and PAF (14/47; 29.8%) treatment groups in the normal male study arm. There was a significant difference in cumulative IUI pregnancy rates between control (10/35; 28.6%) and PAF (14/26; 53.9%) patient groups in the normal male study arm. There was no significant difference in IUI pregnancy rates per cycle between control (12/124; 9.7%) and PAT (14/119; 11.8%) treatment groups in the male factor study arm. There was no significant difference in cumulative IUI pregnancy rates between control (12/46; 26.1%) and PAF (14/38; 36.8%) patient groups in the male factor study arm. There was a significant difference in overall cumulative IUI pregnancy rates between control (21/81; 25.9%) and PAF (27/64; 42.2%) patient groups.Conclusion(s): The inclusion of PAF into the IUI sperm wash procedure significantly improves pregnancy rates. However, the significant improvement can only be shown to affect men presenting with normal semen parameters. (C) 2004 by American Society for Reproductive Medicine.

Objective: Body mass index (BMI) has been demonstrated to impact fertility. Females with a BMI >25 typically are insulin resistant, suffer from PCOS and have a poor fertility prognosis. Those undergoing ART therapy typically require larger gonadotropin dosages and produce fewer mature oocytes plus have a higher incidence of miscarriages. Men with high BMI values present with lower numbers of normal-motile sperm cells. However, the impact of BMI on sperm chromatin integrity is unknown. Therefore the study objective was to determine the relation between BMI and sperm chromatin integrity.Design: Prospective comparison of BMI scores in male patients and sperm chromatin integrity.Methods: Patient height (H) and weight (W) were recorded day of semen collection. BMI was calculated for each patient as follows: (W∗704.5)/(H2). Sperm chromatin integrity was determined by the flow cytometric-based sperm degree of DNA fragmentation chromatin structure assay (SCSA™). Sperm chromatin integrity (the percentage of cells with fragmented DNA) is reported as DNA fragmentation index (DFI).Results: A total of 30 semen specimens were analyzed. The overall BMI mean (+SEM) was 26.9 (+0.62). The overall DFI mean (+SEM) was 24.7 (+2.57). Linear regression [DFI = (1.145∗BMI)-6.079] revealed a significant (P<0.05) and positive relation between BMI and DFI.Conclusion: A positive relationship between BMI and DFI per subject was observed. As men's BMI increases beyond 25 their respective sperm's DFI also increases. Typically, a male presenting with a DFI over 30% will have reduced fertility with an increase of miscarriages. Therefore, infertility patients should be advised to reduce body weight prior to any ART procedure. Additional studies are warranted to determine the impact BMI has upon male fertility and pregnancy outcomes. Objective: Body mass index (BMI) has been demonstrated to impact fertility. Females with a BMI >25 typically are insulin resistant, suffer from PCOS and have a poor fertility prognosis. Those undergoing ART therapy typically require larger gonadotropin dosages and produce fewer mature oocytes plus have a higher incidence of miscarriages. Men with high BMI values present with lower numbers of normal-motile sperm cells. However, the impact of BMI on sperm chromatin integrity is unknown. Therefore the study objective was to determine the relation between BMI and sperm chromatin integrity. Design: Prospective comparison of BMI scores in male patients and sperm chromatin integrity. Methods: Patient height (H) and weight (W) were recorded day of semen collection. BMI was calculated for each patient as follows: (W∗704.5)/(H2). Sperm chromatin integrity was determined by the flow cytometric-based sperm degree of DNA fragmentation chromatin structure assay (SCSA™). Sperm chromatin integrity (the percentage of cells with fragmented DNA) is reported as DNA fragmentation index (DFI). Results: A total of 30 semen specimens were analyzed. The overall BMI mean (+SEM) was 26.9 (+0.62). The overall DFI mean (+SEM) was 24.7 (+2.57). Linear regression [DFI = (1.145∗BMI)-6.079] revealed a significant (P<0.05) and positive relation between BMI and DFI. Conclusion: A positive relationship between BMI and DFI per subject was observed. As men's BMI increases beyond 25 their respective sperm's DFI also increases. Typically, a male presenting with a DFI over 30% will have reduced fertility with an increase of miscarriages. Therefore, infertility patients should be advised to reduce body weight prior to any ART procedure. Additional studies are warranted to determine the impact BMI has upon male fertility and pregnancy outcomes.

Objective: Although essentially a benign disease, endometriosis shares several features with malignancy, including local invasion of tissues and metastasis to distant organs. Thus, the multistep pathway resulting in cellular transformation is considered a valid model to investigate underlying genetic cause in endometriosis. In previous studies, we utilized fluorescent in situ hybridization (FISH) techniques to demonstrate that perturbations of chromosome 17 in general and the p53 locus in particular occur frequently in severe/late-stage endometriosis (Shin et al., 1997; Kosugi et al., 1999). Although FISH provides an attractive approach for single-cell analysis to detect somatic alterations in cells specifically localized to endometriotic lesions, the approach is not only tedious and labor-intensive but also limited by the availability and cost of fluorescently labeled probes. Alternatively, we employed quantitative real-time PCR for more accurate high-throughput assessment of aberrations involving p53 copy number in normal and endometriotic tissue specimens. Design: To examine matched normal blood and endometriotic DNA for quantitative differences in p53 and GAPDH (control locus) copy number compared to normal blood DNA from unaffected women. Methods: Under IRB approval, we obtained fresh surgical tissue specimens and a peripheral blood sample from 22 women undergoing extirpation of advanced stage disease. As a control group, peripheral blood from 28 unaffected women undergoing tubal ligation was also collected. DNA from all tissue specimens was extracted using the QIAamp DNA purification kit. Real-time quantitative PCR (TaqMan Assay; Applied Biosystems) to detect conserved sequences within exon 1 of the p53 gene was performed using the GeneAmp 7700 Sequence Detection System. The ubiquitous autosomal glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locus served as an internal control for PCR efficiency and was used to normalize the values for p53 sequence copies detected. Thus, normalized values reflect the ratio of p53 to GAPDH copies detected. In a normal cell, a normalized value of 1.0 would be expected since both p53 and GAPDH would each contribute two copies. Results: Among control-unaffected cases, normalized mean of 1.003 existed for the p53 locus. No significant differences (P >0.485) in the normalized values for p53 were observed between the unaffected controls and endometriosis subjects (peripheral blood). However, significant (P<0.008) loss or gain of p53 sequences was detected in 17 of 22 (77%) endometriotic tissue specimens. In 13 of the 17 cases, variable gain in p53 sequences was observed (2- to 589-fold greater compared to GAPDH). However, in the remaining 4 cases, loss in p53 sequences was observed, 0.5-fold compared to GAPDH. Conclusion: Our results support the role of non-random somatic alterations involving the p53 locus in the pathogenesis of most cases (77%) of late/sever stage endometriosis. The perturbations could be either gain or loss of p53, consistent with mutation of p53 being not an initial event but a late step. Although the sensitivity of this approach is similar to that of FISH, real-time PCR is semi-automated, allowing for high-throughput evaluation of many more genes and/or chromosomal regions. Therefore, real-time PCR provides an improved method for candidate gene screening at both the DNA and RNA level in endometriosis.

Purpose: Platelet-activating factor (PAF) is a potent signaling phospholipid that is found in mammalian sperm and has a positive correlation with fertility. Whereas PAF is present in human sperm, there are no relational reports on its content and the cells fertilization potential. Therefore, the study objective was to determine if PAF content in capacitated-induced sperm is related to fertilization potential as determined by the sperm penetration assay (SPA).

Endometriosis affects 10–15% of women of reproductive age and is a common cause of infertility and pelvic pain. Although endometriosis is characterized by abnormal growth or turn-over of cells, the genetic changes involved remain unclear. We employed a multi-color fluorescence in situ hybridization (FISH) strategy to determine the incidence of somatic chromosomal numeric alterations in severe/late stage endometriosis. Using alpha-satellite sequence-specific DNA probes for chromosomes 7, 8, 11, 12, 16, 17, and 18, simultaneous two- and three-color FISH were performed to evaluate the frequency of monosomic, disomic, and trisomic cells in normal control and endometriotic tissue specimens. In one of four endometriosis samples studied, a significantly higher frequency of monosomy for chromosome 17 (14.8%, χ24 = 53.3, P < 0.0001) and 16 (8.8%, χ24 = 11.4, P < 0.05) was observed. An increased number of cells with chromosome 11 trisomy (14.8%, χ24 = 96.2, P < 0.0001) were detected in a second case. In a third case, a distinct colony of nuclei with chromosome 16 monosomy (14.1%, χ24 = 21.39, P < 0.005) was detected. Acquired chromosome-specific aneuploidy may be involved in endometriosis, reflecting clonal expansion of chromosomally abnormal cells. That candidate tumor suppressor genes and oncogenes have been mapped to chromosomes 11, 16, and 17 suggests that chromosomal loss or gain plays a role in the development and/or progression of endometriosis.