Optimal utilization of cryopreserved human semen for assisted reproduction: recovery and maintenance of sperm motility and viability.

PURPOSE: Our purpose was to evaluate sperm motility and viability and the maintenance of these parameters in already cryopreserved semen samples following repeated freezing/thawing cycles. METHODS: Human spermatozoa were subjected to five cycles of cryopreservation/thawing. Recovery of sperm motility and viability and the proportion of viable nonmotile sperm were determined up to 6 hr after thaw. RESULTS: Sperm motilities (prefreeze motility, 70.1%; n = 9 samples) after each of five freeze/thaw cycles were 24.4, 8.0, 3.5, 1.5 and 1.8%. The recovery of sperm viability was higher than that of motility after each cycle: 39.1, 25.3, 22.6, 17.8, and 16.5%. Recoveries of motility and viability were improved if the thawed samples were left in the original cryopreservation medium prior to refreezing vs. if a washing/ resuspension step was included. The recovery of sperm motility in the first thawing cycle was indicative of the expected motile sperm recovery in the second thawing cycle. CONCLUSIONS: Cryopreserved semen that is intended to be reused in future assisted reproduction treatments should be thawed only once and aliquoted in the original freezing medium before refreezing. The recovery of sperm motility and viability in the second thawing cycle, thus the applicability of the sample in conventional in vitro fertilization or intracytoplasmic sperm injection may be anticipated in > 90% of the samples. In view of intracytoplasmic sperm injection it is important that sperm viability is maintained better than motility; after the first, second, and third thawing cycles the ratios of motile:nonmotile viable sperm were 1:1, 1:4, and 1:7, respectively.

Improving the intracytoplasmic sperm injection technique by transmembrane electric potential monitoring.

OBJECTIVE: To evaluate electrophysiologic techniques for transmembrane potential measurement during intracytoplasmic sperm injection (ICSI). DESIGN: Mature mouse oocytes were subjected to intracellular measurements of membrane potential using conventional techniques and modifications of the techniques for use with sham ICSI. During the procedure, the actual penetration of the membrane was determined subjectively or objectively with or without monitoring membrane potential. SETTING: Academic medical center. ANIMAL(S): Mouse. INTERVENTION(S): Measurement of oocyte membrane potential (Sham ICSI). MAIN OUTCOME MEASURE(S): Detectability of transmembrane potential using different electrodes and comparison of subjective and objective determination of membrane penetration. RESULT(S): Measurement of the membrane potential with the same glass pipettes used in ICSI means compromise between signal amplitude and compatibility with the conventional ICSI setup. Signal quality is related inversely to the diameter of the injection pipette, and its amplitude decreases as the concentrated electrode filling solution is replaced by physiologic solutions used in ICSI. When successful membrane penetration is suspected by visualization, measuring the potential at the tip of the injection pipette often proves otherwise. Conversely, when membrane penetration is confirmed by detection of transmembrane potential, the procedure may appear subjectively unsuccessful. CONCLUSION(S): Monitoring of transmembrane potential can be done successfully in conjunction with standard ICSI and has several potential applications.

Office Microlaparoscopy Under Local Anesthesia

We assessed the utility, tolerance, and costs associated with office microlaparoscopy under local anesthesia using fiberoptic microlaparoscopes (<2 mm) and accessory instrumentation (<2 mm) for 100 of the following procedures: diagnostic laparoscopy to evaluate infertility including chromopertubation; diagnostic laparoscopy for chronic pelvic pain (CPP), including biopsy of endometriosis and conscious pain mapping; and early second-look laparoscopy including lysis of adhesions. A prospective nonselected cohort study was conducted on all women with CPP or infertility who consented to office microlaparoscopy under local anesthesia. A questionnaire was developed to follow all aspects of patient acceptance and tolerance of the procedures, and women were screened preoperatively, and 30 minutes and 1 week postoperatively. Pain was evaluated with modification of the McGill pain inventory relative intensity scale. A subset of questions evaluated the recovery period, and time and length of all aspects of the procedure and recovery were recorded. Second-look procedures were significantly more painful than diagnostic ones, and women with CPP experienced significantly greater pain than those with infertility. Virtually all expressed high satisfaction with the procedure. There were no complications, and no procedures required general anesthesia. Compared with traditional laparoscopy, there was a reduction in costs greater than 70%.

Baseline cyst formation after luteal phase gonadotropin-releasing hormone agonist administration is linked to poor in vitro fertilization outcome.

OBJECTIVE: To investigate the prognostic significance of baseline ovarian cysts after luteal phase GnRH agonist (GnRH-a) administration for IVF-ET. DESIGN: All nondonor IVF-ET cycles in one program in which luteal phase GnRH-a was administered between July 1993 and January 1994 were assessed for the formation of baseline ovarian cysts defined as a mean diameter > or = 15 mm. Outcome data from the IVF cycles were compared between patients with and without baseline ovarian cysts. RESULTS: Of 78 IVF cycles, baseline cysts > or = 15 mm were noted in 26 cycles. Cycles in which cysts were formed were associated with significantly older patients with significantly higher baseline FSH values. Cycles in which cysts were present demonstrated fewer follicles, retrieved oocytes, and embryos. Cyst cycles also demonstrated a lower peak E2 level, implantation rate, and clinical pregnancy rate (PR) per initiated cycle (7.7% versus 32.7%). Cyst cycles also demonstrated a higher cancellation rate. Logistic regression modeling, accounting for age, confirmed significantly lower clinical PRs in cycles with a baseline cyst. CONCLUSIONS: Baseline cyst formation after luteal phase GnRH-a administration is both a marker for poor responders and a reliable predictor of poor stimulation and low PRs in a given IVF-ET cycle.

Pregnancy in patients with cyclic neutropenia.

Cyclic neutropenia is characterized by periodic episodes of neutropenia that are often associated with infectious complications. A patient with cyclic neutropenia experienced two pregnancies with an amelioration of her infectious complications and an increase in neutrophil count. Cyclic neutropenia may follow a benign course during pregnancy.