Estradiol production during controlled ovarian hyperstimulation correlates with treatment outcome in women undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To study the value of E(2) production during controlled ovarian hyperstimulation (COH) in predicting IVF-ET outcome. DESIGN: Historical cohort. SETTING: Academic infertility center. PATIENT(S): A cohort of 270 patients who completed 324 consecutive IVF-ET treatment cycles. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Area under the curve for E(2) levels (AUC-E(2)) from the first day of COH until the day of hCG administration was calculated and cycles grouped into low, average, and high AUC-E(2) groups. Clinical pregnancy rates per cycle were compared among the three groups, and correlations with AUC-E(2) values were calculated for all patients and after sub-grouping according to age, COH protocol and infertility diagnosis. RESULT(S): Cycles with low and high AUC-E(2) values had significantly lower pregnancy rates particularly in patients 35 years or older. There was a positive correlation between AUC-E(2) and pregnancy rates up to a certain AUC-E(2) level above which a negative correlation was found. The turning point between positive and negative correlations occurred at a significantly lower AUC-E(2) level in patients 35 years or older. CONCLUSIONS: Estradiol production during COH correlates with IVF-ET outcome. Women >35 years of age seem more vulnerable to high E(2) levels.

Area under the curve for estradiol levels do not consistently reflect estradiol levels on the day of hCG administration in patients undergoing controlled ovarian hyperstimulation for IVF-ET.

PURPOSE: Most studies reported estradiol (E2) levels attained on day of hCG administration when investigating effect of E2 on IVF outcome. We studied whether a relationship exists between the area under the curve for E2 levels (AUC-E2) and E2 levels on hCG day during IVF-ET. METHODS: Retrospectively, we analyzed data for 313 patients who completed one IVF-ET cycle each. Patients were sorted according to AUC-E2 levels. Then we compared between each patient's own AUC-E2 and the corresponding E2 level on hCG day for the same patient. RESULTS: Although overall AUC-E2 correlated positively with E2 levels on hCG day, there was no consistent correlation between individual patients. CONCLUSIONS: AUC-E2 reflects more accurately the amount of E2 produced by the follicles during controlled ovarian hyperstimulation. The absence of a uniform correlation between AUC-E2 and E2 on hCG day may result in different conclusions when studying outcomes of IVF treatment.

Live birth following intracytoplasmic sperm injection after a 15-hour weather-related delay in cryopreservation of testicular sperm.

OBJECTIVE: To report a live birth after transfer of cryopreserved blastocysts derived from testicular sperm that were cryopreserved 15 hours after biopsy due to inclement weather. DESIGN: Case report. SETTING: University-affiliated private clinic. PATIENT(S): Couple undergoing IVF with intracytoplasmic sperm injection (ICSI) using testicular extracted sperm for male factor infertility due to unexplained obstructive azoospermia. INTERVENTION(S): An IVF case involving testicular biopsy, cryopreservation, and thawing of testicular tissue, ICSI, embryo freezing, and transfer of cyropreserved blastocysts. MAIN OUTCOME MEASURE(S): The ability to use cryopreserved testicular tissue in which optimal time from biopsy to freezing had been delayed because the weather was inclement. RESULT(S): In spite of a 15-hour delay in processing and cryopreservation of the testicular biopsy, subsequent thawing yielded viable sperm that resulted in viable embryos after ICSI. Transfer of fresh embryos was unsuccessful. However, subsequent use of supernumerary viable blastocysts that were frozen resulted in the live birth of a baby. CONCLUSION(S): A 15-hour delay in processing and cryopreservation of a testicular biopsy did not affect the viability of the extracted sperm after thawing. Although it is desirable to freeze the testicular specimen as soon as possible, the delay in processing in this case due to inclement weather did not affect the potential for fertilization and implantation of a viable cryopreserved blastocyst.

N-Acylethanolamines in human reproductive fluids.

N-Acylethanolamines (NAEs) are an important family of lipid-signaling molecules. Arachidonylethanolamide (anandamide) (AEA), palmitoylethanolamide (PEA), and oleoylethanolamide (OEA) are co-produced from similar phospholipid precursors when neurons are stimulated. AEA is an endogenous agonist (endocannabinoid) for cannabinoid receptors. It binds with higher affinity to type CB1 than to type CB2 cannabinoid receptors. PEA does not bind to CB1, while the hypothesis that it reacts with putative CB2-like receptors has been questioned. OEA does not activate currently known cannabinoid receptors, but it mimics the effects of AEA and cannabinoids in reducing the fertilizing capacity of sea urchin sperm. OEA and PEA also act as entourage compounds by inhibiting the hydrolysis of AEA by fatty acid amide hydrolase. Cannabinoid receptors and/or AEA are present in mammalian reproductive organs including the testis, epididymis, prostate, ovary, uterus, sperm, preimplantation embryo and placenta, as well as prostatic and mammary carcinomas. We now report that analysis by high-performance liquid chromatography/mass spectrometry (HPLC/MS) shows the presence of AEA, PEA, and OEA in human seminal plasma, mid-cycle oviductal fluid, follicular fluid, amniotic fluid, milk, and fluids from malignant ovarian cysts. Previous studies showed that AEA-signaling via cannabinoid receptors regulates capacitation and fertilizing potential of human sperm, early embryonic development and blastocyst implantation into the uterine mucosa of rodents, as well as proliferation of human mammary and prostatic carcinomas. Current results imply that NAEs also may modulate follicular maturation and ovulation, normal and pathological ovarian function, placental and fetal physiology, lactation, infant physiology, and behavior. Collectively, these findings suggest that NAEs in human reproductive fluids may help regulate multiple physiological and pathological processes in the reproductive system, and imply that exogenous cannabinoids delivered by marijuana smoke might impact these processes. This study has potential medical and public policy ramifications because of the incidence of marijuana abuse by adolescents and adults in our society, previously documented reproductive effects of marijuana, and the ongoing debate about medicinal use of marijuana and cannabinoids.

Evidence that anandamide-signaling regulates human sperm functions required for fertilization.

Ejaculated mammalian sperm require several hours exposure to secretions in female reproductive tracts, or incubation in appropriate culture medium in vitro, before acquiring the capacity to fertilize eggs. Arachidonylethanolamide (AEA), also known as anandamide, is a novel lipid-signal molecule that is an endogenous agonist (endocannabinoid) for cannabinoid receptors. We now report that AEA is present in human seminal plasma, mid-cycle oviductal fluid, and follicular fluid analyzed by high-performance liquid chromatography/mass spectrometry. Sperm are sequentially exposed to these reproductive fluids as they move from the vagina to the site of fertilization in the oviduct. Specific binding of the potent cannabinoid agonist [(3)H]CP-55,940 to human sperm was saturable (K(D) 9.71 +/- 1.04 nM), suggesting that they express cannabinoid receptors. R-methanandamide [AM-356], a potent and metabolically stable AEA analog, and (-)delta(9) tetrahydrocannabinol (THC), the major psychoactive constituent of Cannabis, modulated capacitation and fertilizing potential of human sperm in vitro. AM-356 elicited biphasic effects on the incidence of hyperactivated sperm motility (HA) between 1 and 6 hr of incubation: at (2.5 nM) it inhibited HA, while at (0.25 nM) it stimulated HA. Both AM-356 and THC inhibited morphological alterations over acrosomal caps between 2 and 6 hr (IC(50) 5.9 +/- 0.6 pM and 3.5 +/- 1.5 nM, respectively). Sperm fertilizing capacity, measured in the Hemizona Assay, was reduced 50% by (1 nM) AM-356. These findings suggest that AEA-signaling may regulate sperm functions required for fertilization in human reproductive tracts, and imply that smoking of marijuana could impact these processes. This study has potential medical and public policy ramifications because of the incidence of marijuana abuse by adults in our society, previously documented reproductive effects of marijuana, and the ongoing debate about medicinal use of marijuana and cannabinoids.