Stephen Junk

The fertilization of human oocytes by spermatozoa from men with antispermatozoal antibodies in semen

Seventy-two couples, including 15 with antispermatozoal antibodies in the male partners semen, were studied in a program of in vitro fertilization and embryo transfer. Cases were further subclassified as normospermic or oligospermic and antispermatozoal antibodies were assessed with categorization into the respective human immunoglobulin classes as determined using the indirect immunobead test. The study reveals that fertilization is significantly reduced (P<0.001) only if both IgA and IgG antibodies are present in semen but there is no reduction if either class is present alone. The fertilization rate of oocytes is significantly reduced (P<0.001) by sperm from oligospermic samples, and there is a further reduction in those cases with combined IgA/IgG antispermatozoal antibodies.

FSH priming improves oocyte maturation, but priming with FSH or hCG has no effect on subsequent embryonic development in an in vitro maturation program.

AIM To determine whether maturation and subsequent blastocyst development of in vitro matured oocytes can be improved by in vivo follicle stimulating hormone (FSH) or human chorionic gonadotrophin (hCG) priming, using a mouse model. EXPERIMENTAL DESIGN Five groups of oocytes were used: in vivo control, in vitro matured (IVM) control, IVM after 24 h in vivo priming with FSH, IVM after 48 h in vivo priming with FSH and IVM after 16 h in vivo priming with hCG. In vitro fertilization (IVF) was performed on all groups. Oocyte maturation, fertilization, blastocyst development rates and blastocyst cell numbers were assessed for all groups. RESULTS Significant improvement in oocyte maturation was observed in the two FSH priming groups compared with the IVM control group (P<0.005 and P<0.001, respectively). There were no significant differences in fertilization between all five groups. Blastocyst development was significantly higher in the in vivo control compared to the IVM groups (P<0.001). No significant differences were observed in blastocyst cell numbers among all five groups. CONCLUSIONS While FSH priming improves the maturation rate of IVM oocytes, FSH or hCG priming does not improve development to the blastocyst stage.

The successful recovery and fertilization of oocytes from the pouch of Douglas

Two groups of women were studied in whom a proportion of follicles had either ovulated spontaneously (7 women) or ruptured during manipulation at laparoscopy (30 women), and oocytes were recovered from the pouch of Douglas. There were no significant differences in the fertilization rates of oocytes collected in the pouch of Douglas from ovulated follicles, compared with those from the remaining intact follicles [15/20 (75%) vs 14/20 (70%)]. Also there was no significant difference between the fertilization rate of oocytes from follicles ruptured at the time of oocyte collection and that of oocytes from inlact follicles [25/38 (66%) vs 101/140 (72%)]. One woman became pregnant, following the transfer of four four-cell embryos, all derived from spontaneously ovulated oocytes found in the pouch of Douglas. She gave birth to a baby girl. The present study has shown that (1) oocytes may still be retrieved from the pouch of Douglas, despite follicle dispersal; (ii) these oocytes can be fertilized; and (iii) the embryos derived from ovulated oocytes recovered from the pouch of Douglas may generate an ongoing pregnancy following in vitro fertilization and embryo transfer.

The use of latex beads in external quality assurance and internal quality control for routine semen analysis

The usefulness of latex beads of defined concentration was assessed as a substitute for sperm in the performance of External Quality Assurance (EQA) and Internal Quality Control (IQC) of semen analysis. Within the EQA programme, mean±SEM bias (%) was significantly reduced in 2007 compared to 2002 for both specialist (6.0%±5.4% vs. 55.0%±5.9%) and non-specialist (18.4%±5.9% vs. 90.9%±13.4%) laboratories (both p<0.0001), indicating improved accuracy over time. Within the IQC programme, the beads were used in the appraisal of two scientists, one experienced and one inexperienced, against a known standard. Beads were also used to calibrate eleven counting chambers, resulting in one old chamber being discarded due to its poor performance. The present study has shown that the use of a defined concentration of beads is an excellent adjunct to IQC and EQA programmes enabling the performance of both people and equipment to be assessed in an objective manner.

Cytogenetic analysis of embryos generated from in vitro matured mouse oocytes reveals an increase in micronuclei due to chromosome fragmentation.

AbstractPurpose: (i) To determine the prevalence of micronuclei in the cytoplasm of embryos generated from in vitro matured oocytes. (ii) Assess whether micronuclei presence are the result of chromosome fragmentation or the loss of whole chromosomes. Methods: In vitro fertilization was performed on mature oocytes generated from superovulated mice (control) and in vitro matured mouse oocytes. Fertilized oocytes were cultured to the two-cell stage and fixed to slides. Micronuclei assessment was performed after staining with Giemsa. Centromere assessment was made using immunofluorescent staining (CREST) of the centromeric kinetochores. Results: Micronuclei were observed in 2% (4/197) of control two-cell embryos and 36.2% (46/127) of two-cell embryos generated from in vitro matured oocytes (P < 0.02). Centromeres were not detected in micronuclei from either group. Conclusions: A significant increase in micronuclei was observed in embryos generated from in vitro matured oocytes. The lack of accompanying centromeres would suggest the micronuclei are the result of chromosome fragmentation.

Sperm stimulants can improve fertilization rates in male-factor cases undergoing IVF to the same extent as micromanipulation by partial zona dissection (PZD) or subzonal sperm insemination (SUZI): A randomized controlled study

PurposeOur purpose was to evaluate the efficacy of direct insemination (IVF), micromanipulation by partial zona dissection (PZD), and subzonal sperm insemination (SUZI) using spermtreated with pentoxifylline (PF) ± 2-deoxyadenosine (2DA).ResultsThe overall fertilization rate achieved was similar for all three fertilization techniques (33.1, 30.2, and 26.9% for IVF, SUZI, and PZD, respectively). Patients who had reduced fertilization in previous IVF attempts showed improved fertilization with sperm stimulants, either PF alone or PF in combination with 2DA in standard IVF. In certain cases, SUZI or PZD gave significantly improved fertilization rates in comparison to IVF.ConclusionSelective use of sperm stimulants in IVF can achieve fertilization for the majority of male-factor cases. However, PZD and SUZI techniques are useful, especially when sperm stimulants fail to achieve fertilization or achieve poor fertilization in direct insemination.

Are the Australian ART Results as Poor as They Appear

From a once preeminent position in the world of assisted reproduction, the Australian position appears to have slipped into a relatively poor rating on the international scene. Perhaps it might be fairer to say that the comparative data over the past decade show that many nations have advanced their pregnancy rates markedly, while the Australian data indicate a state of minimal progress from a position achieved in the mideighties. Is this really the case? Or is something else happening which our current data presentations fail to reveal? Let us look at both possibilities before drawing conclusions.