Specific isoforms of leucine-rich alpha2-glycoprotein detected in the proliferative endometrium of women undergoing assisted reproduction are associated with spontaneous pregnancy.

OBJECTIVE: To examine differences in specific protein expression from the surface of the human endometrium with respect to eventual pregnancy in infertile women. DESIGN: Laboratory study. SETTING: University hospital. PATIENT(S): Thirty-one women presenting for investigation into infertility at an assisted reproductive unit. INTERVENTION(S): Endometrial flushings were collected during the proliferative phase of the menstrual cycle and subjected to electrophoretic separation on the basis of isoelectric point and molecular weight. Computerized analysis of the resulting spots was performed, and the proteins were identified using tandem mass spectrometry. MAIN OUTCOME MEASURE(S): The expression of individual isoforms of leucine-rich alpha2-glycoprotein (LRG) was compared in nonpregnant patients (n = 25), those who became pregnant as a result of treatment (n = 3), and those who had treatment-independent pregnancies (n = 3). RESULT(S): A statistically significant difference was found in expression of two LRG isoforms, which were higher in the women who subsequently became pregnant independent of treatment. CONCLUSION(S): Several indirect lines of evidence suggest a role for LRG in implantation/decidualization. [1] LRG is implicated in transforming growth factor beta signal transduction. [2] Similar sequences have been identified in murine uterine tissues. [3] LRG may be involved in the infiltration of decidua by uterine natural killer cells, given that the murine homolog of LRG supports lymphocyte infiltration into secondary lymphoid tissues. [4] Human uterine natural killer cells differentiate into granular forms during early pregnancy, and LRG is known to support neutrophil granulocytic differentiation in humans.

Unilateral laparoscopic ovarian diathermy in infertile women with clomiphene citrate-resistant polycystic ovary syndrome.

We evaluated the efficacy of unilateral versus bilateral laparoscopic ovarian diathermy in infertile women with clomiphene citrate-resistant polycystic ovary syndrome. Recall follow-up 18 to 48 months after laparoscopic ovarian diathermy showed that unilateral ovarian diathermy was as effective and long lasting as bilateral ovarian diathermy in the resumption of menstruation and pregnancy rates.

Paternal inheritance of a 16qh-polymorphism in a patient with repeated IVF failure.

Polymorphisms of the size of heterochromatic centromeric regions of chromosomes have been well documented in the human. They appear to have no phenotypic effects in the carriers. However, they appear to be over-represented in infertile couples and those with repeated miscarriages, and there is now growing evidence that they are involved in meiotic pairing, spindle fibre attachment and chromosome movement. Here an analysis of inheritance is reported for a couple presenting with repeated IVF failure in which several embryos were identified as carriers of a polymorphism of the centromeric region of chromosome 16 (16qh-) following aneuploidy screening by sequential fluorescence in-situ hybridization (FISH), using probes for chromosomes 13, 16, 18, 21, 22, X and Y. Detailed cytogenetic analysis by high-resolution banding and FISH of both parents and grandparents established that the polymorphism was familial and inherited from the maternal grandfather. Furthermore, complete analysis of all embryonic nuclei from carrier embryos and others rejected for transfer because of aneuploidy revealed no abnormalities in the segregation pattern of chromosome 16.

Isothermal whole genome amplification from single and small numbers of cells: a new era for preimplantation genetic diagnosis of inherited disease.

Preimplantation genetic diagnosis (PGD) of single gene defects following assisted conception typically involves removal of single cells from preimplantation embryos and analysis using highly sensitive PCR amplification methods taking stringent precautions to prevent contamination from foreign or previously amplified DNA. Recently, whole genome amplification has been achieved from small quantities of genomic DNA by isothermal amplification with bacteriophage 29 DNA polymerase- and exonuclease-resistant random hexamer primers. Here we report that isothermal whole genome amplification from single and small numbers of lymphocytes and blastomeres isolated from cleavage stage embryos yielded microgram quantities of amplified DNA, and allowed analysis of 20 different loci, including the DeltaF508 deletion causing cystic fibrosis and polymorphic repeat sequences used in DNA fingerprinting. As with analysis by PCR-based methods, some preferential amplification or allele drop-out at heterozygous loci was detected with single cells. With 2-5 cells, amplification was more consistent and with 10 or 20 cells results were indistinguishable from genomic DNA. The use of isothermal whole genome amplification as a universal first step marks a new era for PGD since, unlike previous PCR-based methods, sufficient DNA is amplified for diagnosis of any known single gene defect by standard methods and conditions.

Intrauterine insemination results are not affected if Hysterosalpingo Contrast Sonography is used as the sole test of tubal patency.

OBJECTIVE: To assess whether women can be assigned to intrauterine insemination (IUI) treatment on the basis of fallopian tubal patency, diagnosed by Hysterosalpingo Contrast Sonography (Hy Co Sy). DESIGN: Case controlled, clinical study. SETTING: Tertiary referral center. PATIENT(S): The cases were consecutive, infertile women who underwent Hy Co Sy and IUI. The control group was women who had IUI over the same period but whose tubal patency was assessed by hysterosalpingogram (HSG) or laparoscopy and dye (lap and dye). INTERVENTION(S): Women with bilateral patency at Hy Co Sy and with unexplained, anovulatory, or male factor infertility underwent IUI using the partner's (IUI-H) or donor's (IUI-D) semen. Their outcome was compared with that of cohorts of women who had been examined using HSG or lap and dye. MAIN OUTCOME MEASUREMENT(S): Clinical pregnancy rate per cycle and cumulative pregnancy rates at IUI-H or IUI-D. RESULT(S): The clinical pregnancy rates per cycle at IUI-H or IUI-D did not differ among the three groups. The cumulative pregnancy rates after three cycles of IUI-H were 0.17, 0.15, and 0.17 in the Hy Co Sy, HSG, and lap and dye cohorts, respectively, and 0.69, 0.77, and 0.54 in the same groups after six cycles of IUI-D. There were no differences in the clinical characteristics or stimulation regimes used. CONCLUSION(S): Women screened as "normal" by Hy Co Sy may be allocated to treatments that rely on accurate assessment of tubal patency without compromising their chance of conception.