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Objective: To determine whether the process of oocyte vitrification affects oocyte viability in in vitro fertilization (IVF) patients between 30 and 39 years of age., Design: Prospective controlled study., Setting: Private IVF practice., Patient(s): A total of 30 women assigned and 22 qualified., Intervention(s): Denudation of oocytes, cryopreservation of oocytes using vitrification method in a medium with 15% ethylene glycol (EG), 15% dimethylsulfoxide (DMSO), and 0.5 M sucrose., Main Outcome Measure(s): Oocyte survival, fertilization, day-3 embryo quality, blastocyst formation, clinical pregnancy, implantation, and live-birth rates., Result(s): After denudation of oocytes, mature sibling oocytes were randomly allocated to the fresh and vitrified groups. The survival rate was 79.6% after vitrification/warming. Overall, no statistically significant differences were found in fertilization, day-3 embryo quality, or blastocyst formation rates between the fresh and vitrified groups. The positive β-human chorionic gonadotropin, clinical pregnancy rate, and implantation rate were 13 (59.0%) of 22, 10 (45.4%) of 22, and 16 (30.1%) of 53 for the vitrified group. The overall efficiency in achieving a live birth was 11 (5.9%) of 186 per vitrified oocyte., Conclusion(s): The impact of vitrification can be reduced to a minimal level, making it possible to achieve high pregnancy and implantation rates in this age group of IVF patients., (Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

The goal of this work was to select the best combination of encapsulants for the microencapsulation of oregano essential oil by spray dryer with the addition of Arabic gum (AG), modified starch (MS) and maltodextrin (MA). The simplex-centroid method was used to obtain an optimal objective function with three variables. Analytical methods for carvacrol quantification, water activity, moisture content, wettability, solubility, encapsulation efficiency (ME) and oil retention (RT) were used to evaluate the best combination of encapsulants. The use of AG as a single wall material increased ME up to 93%. Carvacrol is the major phenolic compound existent in the oregano essential oil. Carvacrol exhibits a maximum concentration of 57.8% in the microparticle with the use of 62.5% AG and 37.5% MA. A greater RT (77.39%) was obtained when 74.5% AG; MS 12.7% and 12.7% MA were applied, and ME (93%) was improved with 100% of gum.

Objective: To evaluate the efficiency of oocyte donation cycles using egg "cryo-banking.", Design: Study conditions for vitrified/warmed oocytes for 20 non-autologous recipients (from 10 donors) were set prospectively, and outcomes of it were later compared retrospectively to nine fresh donations cycles., Setting: Private assisted reproductive technology program., Patient(s): Ten donors and 20 infertile recipients., Intervention(s): Oocytes were vitrified 3 to 4 hours after collection and cryo-stored. Intracytoplasmic sperm injection was performed 3 hours after warming, and embryos were in vitro cultured for 5 days. Two or three blastocysts were transferred per patient., Main Outcome Measure(s): Oocyte survival, fertilization, development, clinical pregnancy, and implantation rates., Result(s): A total of 153 oocytes were warmed and 134 survived. A total of 117 fertilized and 68% developed to blastocyst stage. A total of 47 embryos were transferred (2.35 embryos per recipient) and 26 implanted. Fifteen patients achieved ongoing pregnancies initially, and two additional pregnancies were obtained after transfer of supernumerary vitrified/warmed embryos. Nine of the 10 donors from the current study had previous fresh donations cycles from where seven clinical pregnancies were established in nine recipients, providing the base for comparison., Conclusion(s): Oocyte donation using vitrified/warmed oocytes can provide high pregnancy and implantation rates, and thus can be considered as efficient treatment procedure with additional benefits to recipients.

Background: Current outcome results with embryos derived from thawed MII human oocyes are significantly lower than with embryos cryopreserved at the pronuclear stage. Here, we investigated whether freezing-thawing was associated with changes in oocyte mitochondrial polarity (DeltaPsim) that could influence competence by altering ATP levels or the ability of the cytoplasm to regulate intracellular Ca2+., Methods: Fresh and thawed uninseminated and unfertilized MII oocytes were stained with the DeltaPsim-specific probe JC-1 to detect clusters of high-polarized mitochondria (J-aggregate positive) and with the Ca2+- specific probe Fluo-4 to measure changes in intracellular levels of this cation. ATP content per oocyte was measured directly and cortical granules were visualized with a cortical granule-specific probe., Results: A significant difference between fresh and thawed MII oocytes existed for pericortical J-aggregate fluorescence and for the ability of the cytoplasm to increase free Ca2+ in response to ionophore exposure. No significant difference in ATP contents was measured and cryopreservation was not associated with an apparent release of cortical granules., Conclusion: Irreversible loss of high DeltaPsim in thawed oocytes may be associated with defects in Ca2+ signalling after insemination and could have downstream consequences for normal embryogenesis., (Copyright 2004 European Society of Human Reproduction and Embryology)

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., Design: Two-cell stage preimplantation stage mouse embryos exposed to PAF (10(-7) M) for 15 min prior to intraoviductal transfer., Methods: Preimplantation stage embryos were recovered from eCG/hCG primed BDF1 female mice. Embryos were exposed to synthetic PAF (10(-7) M) for 15 min. PAF-treated embryos were transferred to the oviducts of pseudopregnant female CD-1 female mice. Superovulated and cultured BDF1 embryos not treated with PAF served as in vitro controls and naturally ovulated embryos with no collection/culture served as in vivo controls. Embryos were permitted to develop to term (18-21 days). The number of pups born per litter and litter weights subsequently were recorded., Results: A total of 160 BDF1 mouse embryos were collected, treated, and transferred (20 per CD-1 recipient) as described. There was a significant (P < 0.05) increase in the number of pups born to the PAF treatment group (56/80; 70%) as compared to the control group (44/80; 55%). There was also a significant difference (P < 0.05) in litter birth weights between the PAF (1.31 g/litter) and controls groups (1.25 g/litter). There was a significant difference (P < 0.05) in birth weights between the PAF treatment group and the in vivo group (1.51 g/litter). There was a significant difference in birth weights between the in vitro-control and in vivo groups (1.51 g/litter). There were no observational malformaties to pups born in any group., Conclusions: Brief exposure of preimplantation stage embryos to PAF will result in a significant increase of delivery rates (pups/litter) as well as birth weights. However, the increase of birth weight was significantly below that found naturally. Additional studies are warranted to elucidate the mechanism of PAF's action in the preimplantation stage embryo and subsequent uterine development.

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 PAF (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.

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)., Methods: Endogenous sperm lipids were measured for PAF content by a specific radioimmunoassay following insemination of zona pellucida-free hamster ova. Data were analyzed by regression analysis and Student's t test., Results: Regression analysis revealed a positive and significant relation (R2 = 0.806; P < 0.05) between PAF content in human sperm and SPA outcome (pass: > or = 5.0; fail: < 5.0, penetrations/ova). Patients that passed (22.61 +/- 5.21 picomoles/10(6)) the SPA had significantly (P < 0.01) higher PAF levels in their sperm than patients that failed (12.91 +/- 1.76 picomoles/10(6) cells) the test., Conclusions: PAF content in capacitated-induced sperm has a significant and positive relationship with fertilization potential. Fertilization potential may be predicted by measuring PAF levels in capacitation-induced human sperm. Determining PAF content in capacitated human sperm may be a beneficial diagnostic tool for the infertility specialist.

Background: A definitive need exists to identify a biomarker of embryonic viability. Platelet-activating factor (PAF) production by human embryos is related to pregnancy potential., Methods: Conditioned embryo culture media were obtained following conventional IVF on day 3, with PAF levels and pregnancy outcomes correlated., Results: Overall pregnancy rate was 68% (17/25) with a mean of 84.1 (+/- 8.5) pmol/l/embryo PAF level. PAF levels ranged from a 216.4 pmol/l/embryo (pregnant) to a 3.7 pmol/l/embryo (not pregnant). There was a significant difference (P < 0.05) in PAF content between pregnant (92.1 +/- 9.5 pmol/l/embryo) and non-pregnant groups (52.5 +/- 16.6 pmol/l/embryo). Patients were categorized into three groups based upon PAF levels: low (< or= 5 pmol/l/embryo); medium (51-100 pmol/l/embryo) and high (>100 pmol/l/embryo). The low (60%) group had a significantly (P < 0.05) lower pregnancy rate than either the medium (85%) or high (89%) groups. A receiver-operator characteristic curve predicted a cut-off limit of 45 pmol/l/embryo for PAF content in human embryo conditioned culture media., Conclusions: The data demonstrate a correlation between PAF levels in human embryo conditioned culture media and pregnancy outcome. Additionally, as embryonic PAF levels increase so does the corresponding pregnancy rate. Therefore, PAF may be used as an indicator of embryo viability and for predicting pregnancy outcome.

Objective: We evaluated the efficacy of blastocyst transfer in decreasing the incidence of high-order multiple gestations after in vitro fertilization., Study Design: We conducted a retrospective analysis of 218 patients who were undergoing in vitro fertilization and by our criteria of three 8-cell embryos on day 3 could receive either a day 3 transfer of cleaved embryos or a day 5 transfer of blastocysts. Ongoing pregnancy rates, implantation rates (determined by the total number of visualized gestational sacs), and multiple pregnancy rates were compared between the 2 groups., Results: Respective day 3 and day 5 ongoing pregnancy rates (61% and 51%) and implantation rates (35% and 33%) were not significantly different. There were 9 triplet or higher gestations in the day 3 group and 0 in the day 5 group., Conclusion: Blastocyst transfer can be used to reduce the number of embryos transferred and the resultant incidence of high-order multiple pregnancies while maintaining high pregnancy rates.

Purpose: Simultaneous fluorescence in situ hybridization (FISH) was used in a preimplantation genetic diagnosis program to determine which embryos were normal for chromosomes X, Y, 13, 18, and 21., Methods: Single blastomeres were disrupted and attached to glass slides using acetic acid and ethanol. Using a ratio mixture of chromosome enumeration DNA probes in combination with locus-specific identifier DNA probes, FISH was performed for the identification of chromosomes X, Y, 13, 18, and 21., Results: Fourteen couples enrolled in IVF produced 134 embryos for biopsy. Blastomeres subjected to five-color FISH revealed that 22% of embryos were normal for chromosomes X, Y, 13, 18, and 21. In addition, 52% were abnormal and no results could be detected for 25%. Twelve couples underwent embryo transfer, two couples did not receive embryos due to lack of any normal embryos, and three couples became pregnant., Conclusions: The simultaneous detection of five-color FISH is a feasible method to detect aneuploidy in preimplantation embryos from women of advanced maternal age.

Objective: Our goal was to achieve a good pregnancy rate after in vitro fertilization; more than one embryo, if available, is transferred to the uterine cavity. This is a recognition of the low implantation rates of embryos from in vitro fertilization. A consequence of this can be high-order multiple implantation with obstetric complications., Study Design: Retrospectively, we reviewed 42 months' in vitro fertilization experience; we related the number of embryos transferred and the pregnancy outcome. During this period 2173 fresh and frozen-thawed embryo transfers were performed. One to six embryos were transferred to women whose average age was 34.4 years (range 21 to 49)., Results: A total of 734 delivered pregnancies (33.8% per embryo transfer) was analyzed according to whether they were single or multiple, and this was related to the original number of embryos transferred. The overall multiple pregnancy rate was 31.3% (24.7% twins, 5.8% triplets, 0.8% quadruplets)., Conclusions: There was a trend toward a higher pregnancy rate with more embryos transferred. The embryonic implantation rate, which reflects the number of embryos that implant per total transferred, was not significantly different in any one group, except in older women in whom more than one embryo was transferred. Whereas greater numbers of embryos (more than three) were transferred in couples with a poorer prognosis for successful in vitro fertilization (e.g., older women [> 36 years old], previous failure of in vitro fertilization, poor embryo quality, or severe male factor causing infertility), there still remained a significant trend toward a higher pregnancy rate when more embryos were transferred. The embryonic implantation rate did not decline in the poorer-prognosis groups (more than three embryos transferred), yet the multiple pregnancy rate was increased. Technologic procedures such as embryo biopsy for aneuploidy screening are proposed as one means to reduce embryo numbers transferred without decreasing the overall pregnancy rate.

Purpose: This retrospective analysis was designed to assess the performance of human embryos following cryopreservation based on whether they were originally developed in standard culture medium (65 cycles, 223 embryos) or cocultured on partial monolayers of bovine oviductal epithelial cells (63 cycles, 198 embryos). Embryo cryosurvival and implantation were compared between the study group and the contemporaneously match controls., Results: During a 2-year period when no factors of the cryopreservation program were altered, 63 transfers of 159 surviving thawed control cleavage-stage embryos (71.3% survival) that were 54% intact gave rise to 11 viable pregnancies (17.5%/ET), to yield an implantation rate of 6.9% per embryo. Sixty-three transfers of 147 thawed cocultured embryos (74.2% survival) that were 61% intact gave an implantation rate of 13.6% per embryo that was significantly higher than the control group (P < 0.05)., Conclusion: Coculture of embryos prior to cryopreservation does not appear to improve cryosurvival; however, it does improve implantation postthaw compared with embryos following standard culture prior to cryopreservation.

Objective: Failure to fertilize eggs in vitro may be countered by micromanipulation of gametes to place selected spermatozoa underneath the zona pellucida of the egg or directly into the egg, thereby improving chances of fertilization and production of viable embryos. Analysis of our clinical data for assisted fertilization was undertaken to assess those factors of relevance in this therapy, and a description of our procedures are given., Study Design: Retrospective analysis of 85 cycles (73 couples) of in vitro fertilization and embryo transfer performed at a private infertility clinic, in which micromanipulation for assisted fertilization was used to overcome either severe male factor infertility or idiopathic failure to fertilize, was performed., Results: In 60 cycles where only embryos from under zona insemination were available for uterine transfer, 15 singleton and two twin pregnancies occurred (28.3% viable pregnancy rate per transfer, 14.1% embryonic implantation). In 14 of these cycles embryos arose only after repeated under zona insemination adding more spermatozoa; this accounted for four of the singleton and one of the twin pregnancies (38.5% pregnancy rate, 22.2% embryonic implantation). No embryos arose from partial zona dissection performed in five cycles on sibling eggs. However, in 16 cycles conventional insemination yielded fertilization in six cycles, and mixed transfer of these embryos and sibling embryos from under zona insemination gave rise to one pregnancy from four transfers (pregnancy rate 25%, embryonic implantation 7.1%). Likewise, in nine cycles donor spermatozoa yielded fertilization in eight cycles, and mixed transfer with sibling embryos fertilized by under zona insemination with partner's spermatozoa gave rise to two pregnancies from five transfers (pregnancy rate 40%, embryonic implantation 15.8%). Fertilization and pregnancy rates did not differ whether couples suffered either from male factor infertility or from previous idiopathic fertilization failure. Direct egg injection of a single spermatozoon into 105 eggs gave an 88.6% egg survival and 32.3% fertilization. Mixed transfers with sibling embryos from conventional and under zona insemination yielded one triplet, one twin, and three singleton pregnancies., Conclusions: Overall, a 24.7% (21/85) viable pregnancy rate per cycle initiated occurred when only embryos from assisted fertilization were available. This strongly indicates that assisted fertilization made a real contribution in cases where either insufficient spermatozoa were available for conventional insemination or in cases where previous fertilization failure had arisen. The wide range of seminal parameters were found to be unhelpful in defining chances of success with assisted fertilization.

Objective: Failure to ejaculate may be overcome by use of electroejaculation. However, such semen samples are often unsuitable for therapies like intrauterine insemination. The combination of electroejaculation with in vitro fertilization, including gamete micromanipulation, should improve chances of fertilization and pregnancy in such cases., Study Design: Within a private infertility clinic electroejaculation in combination with intrauterine insemination was carried out in 18 cycles (10 couples). Four couples went on to receive therapy by electroejaculation plus in vitro fertilization, along with six other couples (15 cycles total) with semen too poor for intrauterine insemination., Results: One term pregnancy arose in the electroejaculation-intrauterine insemination group, and one term pregnancy plus one continuing pregnancy arose from two couples (three cycles) who underwent in vitro fertilization with conventional insemination after electroejaculation. Six couples (nine cycles) had embryos arising only from gamete micromanipulation transferred, and this yielded two term pregnancies, one spontaneous abortion, and a biochemical pregnancy. Two couples (three cycles) failed to achieve fertilization even with micromanipulation; however, donor-inseminated eggs gave rise to two term pregnancies and one continuing pregnancy in these patients., Conclusions: This report confirms the feasibility of in vitro fertilization in conjunction with electroejaculation and extends the therapy to incorporate gamete micromanipulation.

Thirty female New Zealand white rabbits underwent standard laparotomy. Each ovary was bivalved and hemostasis was achieved with bipolar electrocautery. One ovary was then randomized to the closure group, whereas the other ovary was placed in the nonclosure group. In the closure group, the ovarian capsule was closed with a continuous suture of 8-0 Vicryl absorbable surgical suture material with microsurgical technique. In the nonclosure group, the ovaries were left open. Three weeks later the animals were killed and the ovarian adhesions were graded with a standardized scale by an observer (A. A. T.) blinded to the closure status of the animals. Five control animals underwent sham operations with minimal adhesion formation. Statistical analysis of the study animals by the paired Student t test showed a significantly higher adhesion score on the ovaries that were microsurgically closed compared with the ovaries not closed (p = 0.02).

The efficacy of two nonsteroidal antiinflammatory drugs, ibuprofen and indomethacin, in the prevention of postoperative adhesions was examined. Thirty-three guinea pigs were randomly divided into three groups: a control group (n = 11), an ibuprofen group (n = 11), and an indomethacin group (n = 11). All of the animals received standardized injuries, and adhesions were graded 4 weeks later. Both treatment groups were found to have significantly fewer (P less than 0.01) adhesions when compared with the control group with no difference among the two treatment groups. It is concluded that ibuprofen and indomethacin are equally effective in reducing postoperative adhesions.

The Gore-Tex surgical membrane, expanded polytetrafluoroethylene, has been used as a pericardial graft with minimal adhesion formation reported. The purpose of this study was to assess its efficacy as a barrier method for diminishing postoperative peritoneal adhesions in an animal model. Fifteen New Zealand white rabbits underwent laparotomy, with scrape and cut lesions created bilaterally on the uterine body and horns, respectively. On one side, the lesions were covered with the graft using 7-0 Gore-Tex suture; the contralateral side served as an internal control. After four weeks, the adhesions were graded and mean adhesion scores were calculated. The Gore-Tex score was 4.1 times higher than the control for scrape lesions and 1.9 times higher for cut lesions, but the difference was not statistically significant in either case. Overall, the Gore-Tex mean adhesion score was 2.3 times higher than the control, a statistically significant difference. The Gore-Tex surgical membrane did not appear to be an effective adjuvant for postoperative adhesion prophylaxis in this animal model.

The combined experience at two institutions was reviewed to evaluate endometrial biopsy and the prognosis for pregnancies affected by endometrial sampling during the cycle of conception. In the evaluation of 1,084 patients, 1,174 biopsies were performed. Twenty-seven endometrial biopsies were done during the cycle of conception, with spontaneous abortion occurring in six cases (22.2%). Multiple diagnoses were made (including luteal phase defects, endometritis and polyps), and interruption of pregnancy occurred. Several cases illustrate disturbing complications. Methods of minimizing pregnancy interruption and patient discomfort are available.

Unilateral tubal twin pregnancy is exceedingly rare: only 91 cases have been reported in the English literature to date. Two additional such cases are described.

Endometrial tissue from uteri of 35 nonpregnant, premenopausal women was assayed for prostaglandin E2 and F2 alpha binding site content as a function of the phase of the menstrual cycle and the pathologic state. For all specimens, tritium-labeled prostaglandin F2 alpha, binding was very low (less than 8 fmol/mg of protein) or undetectable regardless of the phase of the menstrual cycle or pathologic state or in the presence or absence of 10 mumol/L of indomethacin, a prostaglandin synthetase inhibitor. However, tritium-labeled prostaglandin E2 binding was detected in every specimen and was independent of the presence or absence of indomethacin. Binding of tritium-labeled prostaglandin E2, as determined by Scatchard analyses, was biphasic (dissociation constant approximately 1 nmol/L; dissociation constant for low-affinity sites approximately 10 nmol/L) for both proliferative and secretory endometrial tissue. However, the total number of prostaglandin E2 binding sites, determined from Scatchard or single-point analyses, was significantly higher (p less than 0.01) in proliferative endometrium compared to secretory endometrium. In addition, for endometrium from the proliferative phase of the menstrual cycle, the diagnosis of abnormal uterine bleeding was associated with higher (p less than 0.01) tritium-labeled prostaglandin E2 binding than diagnosis of dysmenorrhea, stress urinary incontinence and uterine prolapse, or pelvic inflammatory disease. Endometrial specimens with the last four diagnoses did not differ significantly (p greater than 0.1) from each other.

During the 12-month study interval ending March 30, 1986, there were 203 ectopic pregnancies at Grady Memorial Hospital, a ratio of one ectopic gestation per 34 deliveries. Twenty patients with isthmic ectopic pregnancies were selected for treatment by one of three operative modalities. Seven patients with ruptured isthmic ectopic pregnancies underwent segmental tubal resection without reanastomosis. All four patients who underwent segmental tubal resection with primary microsurgical reanastomosis had postoperative hysterosalpingograms demonstrating bilateral tubal patency. One pregnancy has occurred in this group. Nine patients underwent linear salpingostomy. In five of the six patients who had postoperative hysterosalpingography, patency of the involved fallopian tube was demonstrated. Four of these nine patients, including one patient with contralateral tubal occlusion, have conceived. We conclude that linear salpingostomy for isthmic ectopic pregnancies is as effective as segmental tubal resection with primary microsurgical reanastomosis in achieving tubal patency.