Oolemma rupture inside the intracytoplasmic sperm injection needle significantly improves the fertilization rate and reduces oocyte damage.

OBJECTIVE: To compare the effect on fertilization, oocyte damage, embryo freezing, and pregnancy rates of two different techniques for rupturing the oolemma during intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective study. SETTING: Fertility Center, Alliant Health System Hospital. PATIENT(S): Seventy-nine consecutive IVF-ICSI cases. INTERVENTION(S): Patients in group I had ICSI performed by pushing the needle into the oocyte until the oolemma was observed to break outside the needle. In group II the oolemma was aspirated into the needle until it ruptured inside the needle. MAIN OUTCOME MEASURE(S): In group II ICSI resulted in significantly higher fertilization and lower oocyte damage rates (66% and 13%) than in group 1 (39% and 29%). There were no statistically significant differences in embryo cleavage rates or pregnancy rates per retrieval between the two groups. A greater number of cases had embryos cryopreserved in group II than in group I. RESULT(S): Rupturing the oolemma by aspirating it into the ICSI needle (group II) improved laboratory outcomes compared with the more traditional technique of breaking this membrane by the stabbing action of the needle (group I). This modification of the ICSI technique also increased the number of patients with cryopreserved embryos and therefore could increase the pregnancy rate per patient. CONCLUSION(S): The site and technique used to rupture the oolemma during ICSI has a significant effect on the fertilization and damage rates.

Improved clinical outcomes for in vitro fertilization with delay of embryo transfer from 48 to 72 hours after oocyte retrieval: use of glucose- and phosphate-free media.

OBJECTIVE: To evaluate clinical outcomes of day 2 versus day 3 ET using a culture media with no glucose or phosphate. DESIGN: Retrospective clinical study. SETTING: Hospital-based fertility clinic. PATIENT(S): One hundred seventy-six IVF-ET patients undergoing controlled ovarian supraovulation. INTERVENTION(S): IVF and delaying the ET by 1 day. MAIN OUTCOME MEASURE(S): Number of blastomeres per embryo, implantation and pregnancy rates. RESULT(S): Delaying the ET from day 2 to day 3 after oocyte retrieval significantly increased implantation rates (13% versus 24%) and ongoing/delivered pregnancy rates per retrieval (26% versus 44%). Day 3 embryos with > or = 8 blastomeres resulted in a significantly higher pregnancy rate (53%) than day 3 embryos with < 8 cells (23%) and day 2 embryos with > or = 4 cells (31%) or < 4 cells (11%). CONCLUSION(S): Day 3 ET was associated with a significant increase in implantation and pregnancy rates. Delaying the ET until day 3 may permit the selection of more viable embryos than on day 2. The absence of glucose and phosphate from the culture media is compatible with good IVF outcomes.

A simplified in vitro assay of delayed hypersensitivity in diagnosis of dermatomycoses.

A tissue culture technique for detection of cellular hypersensitivity in an animal model of dermatomycoses is presented. It is based on specific inhibition of migration of leukocytes sensitized to dermatophytic antigens. The application and specificity of this new immunological technique were studied in guinea pigs infected with Trichophyton rubrum, Microsporum vanbreuseghemii, and Epidermophyton floccosum. In all the infected animals, sensitivity to trichophytin was demonstrated to be independent of the clinical disease phase. The in vivo skin-test reactions were common to those of the group of dermatophytes and did not distinguish between the different species. A statistical difference was observed in the leukocyte migration indices of sensitized cells to the homologous and the heterologous antigens of these pathogens (p less than 0.05, Student's test). It is concluded that the leukocyte migration inhibition assay provides a specific expression of cellular hypersensitivity and may be considered suitable for investigation of cellular immunity in vitro in clinical diagnosis or research. This is a simple, rapid, and inexpensive technique that requires small volumes of peripheral blood.