Fertilization and pregnancies following intracytoplasmic injection of testicular spermatozoa

Purpose Intracytoplasmic injection (ICSI) with testicular sperm was performed in 16 couples. All men had ductal obstruction and failed previous attempts of epididymal sperm microaspiration.Methods Testis tissue was obtained by excisional biopsies and incubated in HEPES buffered EBSS medium over 24 h at 37C. Motile sperm (Grade 1 to 2) were recovered in 13 patients and fertilized a total of 62 oozytes. Four pregnancies were achieved.Results One healthy boy and two girls (twin pregnancy) were born.Conclusions The ongoing pregnancies revealed no fetal abnormalities on ultrasound scanning.Presented at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, April 3–7, 1995, Vienna, Austria.     

Novel method for the cryopreservation of testicular sperm and ejaculated spermatozoa from patients with severe oligospermia: a pilot study

OBJECTIVE: To investigate Volvox globator as an easy-to-handle vehicle and as a safe alternative for cryopreservation of functional motile sperm cells. DESIGN: Prospective, controlled, clinical pilot study. SETTING: Two in vitro fertilization (IVF) outpatient clinics for reproductive medicine. PATIENT(S): Fifteen patients with severe male infertility (density <100 motile sperm per milliliter) who were recruited from two IVF programs. The sperm cells were not intended for clinical use after thawing. INTERVENTION(S): In each case, a predetermined number (n = 8) of motile and morphologically intact sperm cells were injected into each Volvox sphere and then cryopreserved. The quality of the sperm cells and the handling of the Volvox spheres were verified. MAIN OUTCOME MEASURE(S): Postthaw recovery rate in cases of severe male infertility and the amount of motile sperm after thawing. RESULT(S): The postthaw recovery rate was 100%. At least 60% of the sperm cells were motile after thawing. CONCLUSION(S): The use of the spherical algae Volvox globator offers a promising, inexpensive, and easy approach to the cryopreservation of functional motile sperm cells. Volvox globator is an alternative in countries that prohibit the destructive use of oocytes, even after fertilization has failed.     

Ultrastructural observations in human oocytes and preimplantation embryos after zona opening using an erbium-yttrium-aluminium-garnet (Er:YAG) laser

For more than 3 years we have performed laser-assisted hatching prior to embryo transfer in patients with recurrent implantation failure using an erbium-yttrium-aluminium-garnet (Er:YAG) laser system that operates in the infrared region of the light spectrum. The laser beam is guided through a quartz fibre and is brought into direct contact with the zona pellucida. This study was undertaken to evaluate the ultrastructural effects of this laser on the zona pellucida and underlying cell membrane of unfertilized human oocytes and pathologically fertilized preimplantation embryos using light and scanning electron microscopy. The Er:YAG laser produces an almost circular zona opening in the shape of a truncated cone tapering off towards the inside, with a mean diameter of 18 mm. The exact diameter of the drilled site depends on the diameter of the fibre tip and the total number of pulses applied. After laser interaction, the zona matrix and the surface of the underlying ooplasm membrane showed no degenerative alterations. We conclude that the Er:YAG laser is an effective microsurgical tool for achieving reproducible, precise zona openings particularly suitable for purposes of assisted hatching because of their characteristic shape.      

Axillary metastases of an occult primary carcinoma of the breast-discovered only by 99mTc-tetrofosmin scintigraphy

Recent reports consider 99mTc-tetrofosmin scintigraphy to be a powerful new diagnostic tool for discriminating malignant from benign breast disease. We report on a woman suffering from histologically confirmed axillary metastases of a primary unknown, occult carcinoma, whose origin was suspected within the breast. All the diagnostic procedures performed to discover any lesion failed or were inconclusive. The primary cancer was clearly visualized, however, in the right breast by means of 99mTc-tetrofosmin scintigraphy. Conclusion. We suggest that 99mTc-tetrofosmin scintigraphy is a powerful method to detect breast cancer, especially when other diagnostic imaging procedures are inconclusive.     

Non-invasive detection of alterations of the carotid artery in pregnant women with high-frequency ultrasound.

OBJECTIVE: To compare the thickness of the layers of the carotid artery wall in pregnant and fertile non-pregnant women. DESIGN: Prospective cross-sectional study. SUBJECTS: Fifty-one pregnant women at a mean gestational age of 38.9 weeks and 64 fertile non-pregnant women were examined at a University hospital. METHODS: The three layers (adventitia, media, intima) of the superficial wall of the left common carotid artery were identified and measured with high-resolution ultrasound (22.5 MHz). RESULTS: Pregnant women had a thinner intima layer (0.25 +/- 0.07/0.29 +/- 0.08 mm) and a thicker media layer (0.31 +/- 0.08/0.27 +/- 0.09 mm) compared with controls. A statistically significantly higher intima/media ratio was calculated for the pregnant women (1.14 +/- 0.03), compared with the non-pregnant women (0.88 +/- 0.04). CONCLUSION: There are differences in the thickness of the histological layers of the carotid artery wall in pregnant compared with non-pregnant women. This is likely to be due to the effect of different estradiol levels in these two groups.     

Premature ovarian failure: etiology and prospects.

A search of past and current articles on ovarian physiology and premature ovarian failure (POF) using MEDLINE was performed in order to present an overview of clinical manifestations, necessary laboratory investigations, possible etiologies and treatments for POF. POF is defined as gonadal failure before the age of 40 years. Initially, POF was thought to be permanent, but it is now believed that spontaneous remissions and even pregnancies are possible in affected women. In most cases, the etiology of POF remains elusive, but several rare specific causes have been identified. Although the etiology of POF is heterogenic, the treatment principles are the same. Hormone replacement therapy (HRT) is still the cornerstone of treatment. The only proven method of obtaining a pregnancy in patients with POF is fertilization of a donor oocyte. Cryopreservation of oocytes has worked well in animals but awaits refinement before it can be applied routinely to humans with prodromal POF, or to patients before chemotherapy or irradiation in order to save their oocytes for future fertilization. New alternatives to traditional HRT and methods of fertility preservation are under development, but understanding of the basic pathophysiology of POF is necessary for the development and use of innovative treatments.     

Prospective randomized study of an ultrashort gonadotrophin-releasing hormone agonist versus a modified suppression protocol for ovarian stimulation in intracytoplasmic sperm injection cycles

To compare oocyte quality and clinical outcome after an ultrashort or a modified suppression gonadotrophin-releasing hormone agonist (GnRHa) protocol for ovarian stimulation in intracytoplasmic sperm injection (ICSI) cycles, we conducted a prospective randomized study of 60 consecutive couples with severe male infertility admitted for their first in-vitro fertilization (IVF) and ICSI attempt. More cycles were cancelled after the ultrashort protocol (8/30) than after the modified suppression protocol (3/30), although the difference was not significant. There were no cases of severe ovarian hyperstimulation syndrome (OHSS) in the ultrashort group compared to three cases in the suppression group. The percentage of mature metaphase II oocytes recovered in both groups was similar (88 versus 86%), as were the fertilization or cleavage rates after ICSI. In the ultrashort group, a total of 64 embryos was replaced in 22 transfers (mean 2.9 embryos per transfer), resulting in three first trimester abortions and seven deliveries. In the suppression group, 11 deliveries were achieved after transfer of a total of 75 embryos in 27 patients (mean 2.8 embryos per transfer). In conclusion, there was no apparent difference between the two GnRHa protocols in terms of oocyte quality and clinical outcome. However, because of the lower rate of severe OHSS, in our study the ultrashort protocol was more appropriate for ovarian stimulation in ICSI cycles than the modified suppression protocol.      

Cetrorelix in an oral contraceptive-pretreated stimulation cycle compared with buserelin in IVF/ICSI patients treated with r-hFSH: a randomized, multicentre, phase IIIb study

BACKGROUND: The aim of this study was to assess the non-inferiorityof an oral contraceptive (OC)-pretreated cetrorelix regimenand a buserelin regimen in IVF/ICSI patients treated with r-hFSHin terms of total number of oocytes retrieved. METHODS: Multicentre,randomized study. One hundred and eighty two patients were randomizedto receive cetrorelix with OC pretreatment (n = 91) or to receivebuserelin (n = 91). The cetrorelix group started with dailyOCs on cycle day 5 and continued for 21–28 days. Cetrorelix(0.25 mg) was given daily from stimulation day 6 up to and includingthe day of r-hCG administration. The buserelin group startedwith buserelin (500 µg/day) for at least 10 days untildown-regulation was achieved, after which the dose was reducedto daily 200 µg up to and including the day of r-hCG administration.r-hFSH was started in both groups on a Friday, in the cetrorelixgroup 5 days after the last OC pill intake. Both regimens werefollowed by a standard IVF or ICSI procedure. The primary efficacyendpoint was the number of oocytes retrieved per patient. RESULTS:Number of oocytes, cancellation rates, r-hFSH requirements,number of oocyte retrievals during the weekend or public holidayand number of pregnancies were similar in both groups. Bothtreatment regimens were well tolerated. CONCLUSIONS: Cetrorelixpretreated with OCs resulted in similar number of oocytes retrievedcompared with a long buserelin protocol. Both regimens werewell tolerated and allowed scheduling of the oocyte retrieval,with only small number of retrievals falling on a weekend orpublic holiday.     

A new method to process testicular sperm: combining enzymatic digestion,accumulation of spermatozoa,and stimulation of motility

Purpose

In azoospermia processing of the TESE material often results in a sample of reduced purity. This prospective study was set up to clarify whether a combination of enzymatic digestion, density gradient centrifugation and stimulation of motility (where indicated) is a feasible option in TESE patients.

Methods

A total of 63 samples (showing spermatozoa) were processed by the present tripartite processing method. The resulting sperm sample of high purity was directly used for ICSI and subsequent cryopreservation when quality of the accumulated sperm sample allowed for it (n = 39 cycles).

Results

Compared to the control group blastocyst formation rate in the present tripartite processing technique was significantly (P < 0.01) higher (55.2 vs. 43.7 %). Fertilization rates differed significantly (P < 0.001) between cases in which motile sperm could be used (58.4 %) compared to ICSI with immotile sperm (45.0 %). Clinical pregnancy rate per transfer was 40.0 % (24/60) using fresh and 21.6 % (8/37) with cryopreserved TESE material. The calculated live birth rates were 31.7 and 21.6 %, respectively. Thirty-five healthy children were born.

Conclusions

A comparison with a control group suggests that the present approach using standardized ready-to-use products is efficient and reliable. Presumably healthy live births further indicate the safety of the procedure.