PIEZO-ICSI increases fertilization rates compared with conventional ICSI in patients with poor prognosis.

PURPOSE: Limited research has been published comparing PIEZO-ICSI with conventional ICSI. While positive effects have been documented in improving fertilization and degeneration, the outcomes in patients with previous poor results from conventional ICSI remain unclear. It is hypothesized that these patients may benefit the most from this form of insemination. METHODS: This retrospective paired within-patient cohort study investigated patients (n=72) undertaking PIEZO-ICSI after a previous conventional ICSI cycle resulted in poor outcomes (including low fertilization (<50%), high degeneration (>15%), and/or poor embryo development and utilization). Patients required at least five oocytes collected in both cycles and a period of less than 2 years between the cycles. The outcomes of both cycles were compared in respect to fertilization, degeneration, embryo utilization, and pregnancy rates. Further analyses were applied to patients <38 and ≥38 years of age, with <50% or ≥50% fertilization with conventional ICSI and with <20% or ≥20% utilization with conventional ICSI. RESULTS: PIEZO-ICSI resulted in significantly higher fertilization (61.9% vs 45.3%, P<0.0001) and lower degeneration (7.7% vs 18.2%, P=0.0001) when compared to the conventional ICSI cycles. The greatest benefit was seen in patients who had less than 50% fertilization or <20% utilization in their conventional ICSI cycle, with improvements in fertilization and degeneration rates resulting in a significantly higher number of embryos utilized (frozen or transferred) per cycle. CONCLUSIONS: PIEZO-ICSI improved fertilization, degeneration, and utilization rates in patients with previous poor outcomes from conventional ICSI. The number of embryos available for use per cycle was also increased. Further significant improvements were achieved in patients who exhibited poor fertilization (<50%) or low utilization (<20%) from conventional ICSI.

Endometrial stem/progenitor cells in menstrual blood and peritoneal fluid of women with and without endometriosis.

RESEARCH QUESTION: Are endometrial stem/progenitor cells shed into uterine menstrual blood (UMB) and the peritoneal cavity in women with and without endometriosis during menstruation? DESIGN: Women with (n = 32) and without endometriosis (n = 29) at laparoscopy (total 61), carried out during the menstrual (n = 41) and non-menstrual phase (n = 20) were recruited. The UMB, peritoneal fluid and peripheral blood were analysed by clonogenicity assay and flow cytometry to quantify the concentrations of endometrial clonogenic cells, SUSD2+ mesenchymal stem cells (eMSC) and N-cadherin+ epithelial progenitor cells (eEPC). RESULTS: Clonogenic endometrial cells, eMSC and eEPC were found in most UMB samples at similar concentrations in women with and without endometriosis. In contrast, 62.5% of women with endometriosis and 75.0% without (controls) had clonogenic cells in peritoneal fluid samples during menses. The eMSC were present in the peritoneal fluid of 76.9% of women with endometriosis and 44.4% without, and eEPC were found in the peritoneal fluid of 60.0% of women with and 25.0% without endometriosis during menses. Median clonogenic, eMSC and eEPC concentrations in peritoneal fluid were not significantly different between groups. More clonogenic cells persisted beyond the menstrual phase in the peritoneal fluid of women with endometriosis (menstrual 119/ml [0-1360/ml] versus non-menstrual 8.5/ml [0-387/ml]; P = 0.277) compared with controls (menstrual 76.5/ml [1-1378/ml] versus non-menstrual 0/ml [0-14/ml]; P = 0.0362). No clonogenic endometrial cells were found in peripheral blood. CONCLUSIONS: Clonogenic endometrial cells, SUSD2+ eMSC and N-cadherin+ eEPC are present in UMB and the peritoneal fluid of women with and without endometriosis. Further study of the function of these cells may shed light on the cellular origins of endometriosis.

Cryopreserved oocytes: update on clinical applications and success rates.

IMPORTANCE: Over the past 3 decades, oocyte cryopreservation procedures have improved rapidly. However, there is limited research reviewing the efficacy of different cooling protocols and inadequate data comparing in vitro fertilization (IVF) outcomes from fresh oocytes with cryopreserved oocytes. OBJECTIVE: The present review was performed to investigate advances in oocyte cryopreservation technologies and identify areas for further research, to determine whether results from IVF using cryopreserved oocytes are comparable to IVF using fresh oocytes, and to identify the patient populations requiring access to oocyte cryopreservation. EVIDENCE ACQUISITION: A literature review was conducted. OVID (MEDLINE) and PubMed databases were queried using phrases such as "oocyte or egg" and "cryopreservation," "vitrification," or "slow cooling or slow freezing." A total of 180 studies were selected for review. RESULTS: Current literature suggests that vitrified oocytes produce superior IVF results to slow-frozen oocytes and may yield comparable outcomes to IVF with fresh oocytes in certain patient populations. Patients at risk of infertility due to disease or age-related decline or oocyte donation programs, couples who fail to produce semen when required for IVF, and patients with legal or ethical reasons against embryo cryopreservation may access cryopreserved oocytes. CONCLUSIONS: We suggest that women who comprise the previously mentioned patient populations should be offered oocyte vitrification technology. Further research is required to confirm IVF success across all patient populations and determine the best cryopreservation protocols. RELEVANCE: This review will be relevant to clinicians interested in fertility treatments using cryopreserved oocytes, fertility preservation, oncology and fertility, and immunology and fertility.

Utilisation of ART in single women and lesbian couples since the 2010 change in Victorian legislation.

Enactment of the Assisted Reproductive Treatment Act (Vic) 2008 in January 2010 allowed single persons and same sex couples in Victoria to access reproductive treatments. A retrospective cohort analysis of Monash IVF patients was conducted to identify trends in Assisted Reproductive Technology (ART) use amongst single women and lesbian couples after January 2010. A 102.8% increase in the utilisation of ART was observed amongst the single women group and a 248.8% increase in the lesbian couple population.

Interleukin (IL)11 mediates protein secretion and modification in human extravillous trophoblasts.

BACKGROUND: Human trophoblast invasion and differentiation are essential for a successful pregnancy outcome. Dysregulation of these processes can lead to placental pathologies such as pre-eclampsia. The molecular mechanisms; however, are poorly understood. Interleukin (IL)11--a cytokine that regulates endometrial epithelial cell adhesion, trophoblast motility and invasion during implantation--may be involved in some of these processes. METHODS AND RESULTS: The effect of IL11 on protein expression was investigated in trophoblastic HTR8/SVneo cells and primary extravillous trophoblasts (EVTs) purified from first- trimester placentas. Two-dimension (2D)-differential in-gel electrophoresis analyses revealed that 731 spots were significantly differentially regulated by IL11 in HTR8/SVneo cells: seven spots were analyzed by liquid chromatography-tandem mass spectrometry and 14 unique proteins identified. Protein disulfide isomerase family A, member 3 (PDIA3; endoplasmic reticulum p57) and glucose-regulated protein 78 (GRP78) were further validated to be regulated by IL11 in HTR8/SVneo and primary EVT. One dimension western blot analysis confirmed that PDIA3 was down-regulated in EVT. 2D western blot analysis revealed that GRP78 was post-translationally modified following IL11 treatment. Moreover, IL11 stimulated the secretion of GRP78 in EVT. CONCLUSIONS: Data suggest that IL11, possibly via signal transducers and activators of transcription 3 signaling pathway, regulates PDIA3 protein expression and modification/secretion of GRP78. This is the first study to identify PDIA3 and GRP78 as IL11 targets in invasive trophoblasts and identifies a possible mechanism by which IL11 regulates trophoblast function.

The effects of chemotherapy and radiotherapy on fertility in premenopausal women.

UNLABELLED: With the improved survival rate of childhood and young adult cancer patients, the long-term sequelae of the treatments used are increasingly important. In this review, current knowledge of the gonadotoxicity of commonly employed chemotherapeutic agents and radiotherapy regimens is examined. Differences between the effect of "high-risk" and "low-risk" agents are discussed. Tailoring treatment to suit the individual and counseling patients regarding reduced fertility have resulted in the best practice. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completing this CME activity, physicians should be better able to evaluate and use appropriate methods to estimate ovarian reserve, assess the risk of infertility caused by commonly used cytotoxic chemotherapy regimens and radiation, and counsel patients regarding the gonadotoxic effects of cancer treatment.

In vitro fertilisation (IVF) with donor eggs in post-menopausal women: are there differences in pregnancy outcomes in women with premature ovarian failure (POF) compared with women with physiological age-related menopause?

PURPOSE: This study assessed pregnancy rates and obstetric outcomes in women with premature ovarian failure (Group A) with post-menopausal women > or =40 years (Group B) who had IVF +/- ICSI using donor eggs. METHODS: This was a retrospective analysis of 54 recipients with either premature ovarian failure or physiological menopause undergoing oocyte donation between 2000 and 2007 at Monash IVF. RESULTS: The average number of stimulated cycles required for a woman in group A and B to deliver a baby was 1.75 and 1.4 respectively. Both groups had high cumulative pregnancy rates; however, there was a statistically significant difference with regards to rates of complications. CONCLUSION: Oocyte donation in both premature ovarian failure and physiological menopause is highly successful and cumulative pregnancy rate is an important statistic which can be used to inform women seeking this technique. High rates of complications, in conjunction with individual risk-factor analysis needs to be considered when counselling post-menopausal women about oocyte donation.

Blastocyst transfer does not cause a sex-ratio imbalance.

OBJECTIVE: To determine whether either single or double fresh blastocyst transfers result in a sex-ratio imbalance in resulting offspring compared with transfers on day 2 or 3 and whether there is a correlation between rate of embryo development and sex of the embryo. DESIGN: Retrospective analysis. SETTING: Large IVF center. PATIENT(S): Four hundred thirty-five live births from single fresh ETs for the period January 2005 through July 2007 and 2,043 live births from double ETs for the same period. INTERVENTION(S): Statistical analysis performed on sex ratio of offspring resulting from transfers (day 2, day 3, day 4, and blastocyst), as well as on the stage of development reached for each day in culture analyzed on sex of the embryo. MAIN OUTCOME MEASURE(S): Sex ratio of offspring by day of transfer. Stage of embryonic development by sex for each day in culture. RESULT(S): There was no difference in sex ratio with blastocyst transfer (single or double). There was no difference in speed of embryonic development at any stage in vitro. CONCLUSION(S): Male embryos do not grow faster than female embryos in culture. Blastocyst transfer does not result in a sex-ratio imbalance in resulting offspring.

Pathophysiology of fibroid disease: angiogenesis and regulation of smooth muscle proliferation.

Uterine fibroids are the most common tumours presenting in women. The pathophysiology of fibroids is poorly understood, but disordered angiogenesis and altered smooth muscle cell proliferation are believed to play a role. In this review, current knowledge of both of these processes will be summarized. Differences between 'normal' adjacent myometrium and fibroid tumours within the same uterus are outlined. Exploiting these differences represents one of the best opportunities for the development of medical treatments that target fibroid tissue selectively.

Molecular profiling of human endometrium during the menstrual cycle.

This summary of the findings of investigations of the changing transcriptional profile of human endometrium during the menstrual cycle shows that it is possible to classify the menstrual cycle based on the global gene expression profile, and identifies groups of known and novel genes that may be associated with different biological processes that occur in the endometrium such as implantation and menstruation.

Anatomical evidence for in utero androgen exposure in women with polycystic ovary syndrome.

OBJECTIVE: To determine whether women with polycystic ovary syndrome (PCOS) have masculinized finger length patterns compared to women without PCOS. DESIGN: A case control study. SETTING: University teaching hospital and in vitro fertilization unit. PATIENT(S): Seventy women aged between 18 and 40 years with PCOS were compared to 70 women without PCOS. INTERVENTION(S): Measurement of the second to fourth finger length ratio on the ventral surface of the left and right hand from the basal crease of the digit to the tip was made using Vernier calipers. MAIN OUTCOME MEASURE(S): The second to fourth finger length ratio. RESULT(S): We found a significantly reduced ratio in the right hand of the women with PCOS compared to the controls. The geometric mean right finger length ratio in the PCOS group was 98.3% that of the controls (95% confidence interval, 99.3%-97.3%). CONCLUSION(S): Here we show a subtle difference in the finger length pattern of women with PCOS. This may constitute anatomical evidence of in utero androgen exposure in PCOS.

Office ultrasound should be the first-line investigation for confirmation of correct ESSURE placement.

BACKGROUND: Hysteroscopic options for permanent birth control (PBC), such as the ESSURE device, are becoming increasingly popular as an alternative to laparoscopic tubal ligation. The success of the technique hinges upon correct device placement within the intramural portion of the fallopian tube. OBJECTIVE: To determine the utility of office ultrasound for confirming correct ESSURE PBC device placement at the 3-month check in a general gynaecology practice. STUDY POPULATION: The first 99 patients in a single centre following ESSURE PBC device placement. TYPE OF STUDY: Prospective cohort study. METHODS: Clinical data was reviewed from patient records, both from the time of the initial procedure and from the follow-up at 3 months. All women underwent an ultrasound at the 3-month check. RESULTS: The ESSURE PBC devices were placed successfully in 84.8% of cases. Of those cases with apparently successful placement, office ultrasound alone confirmed correct device placement at the 3-month check in 94% of cases. Further imaging was needed in only 6% of cases. DISCUSSION: Office ultrasound performed by the general gynaecologist at the 3-month check is more convenient for the patient, and is sufficient to confirm ESSURE PBC device placement in the vast majority of cases. We propose that the protocol for ESSURE PBC device follow-up should be altered to replace X-ray with ultrasound as the first-line investigation.

Bioinformatic analysis of primary endothelial cell gene array data illustrated by the analysis of transcriptome changes in endothelial cells exposed to VEGF-A and PlGF.

We recently published a review in this journal describing the design, hybridisation and basic data processing required to use gene arrays to investigate vascular biology (Evans et al. Angiogenesis 2003; 6: 93-104). Here, we build on this review by describing a set of powerful and robust methods for the analysis and interpretation of gene array data derived from primary vascular cell cultures. First, we describe the evaluation of transcriptome heterogeneity between primary cultures derived from different individuals, and estimation of the false discovery rate introduced by this heterogeneity and by experimental noise. Then, we discuss the appropriate use of Bayesian t-tests, clustering and independent component analysis to mine the data. We illustrate these principles by analysis of a previously unpublished set of gene array data in which human umbilical vein endothelial cells (HUVEC) cultured in either rich or low-serum media were exposed to vascular endothelial growth factor (VEGF)-A165 or placental growth factor (PlGF)-1(131). We have used Affymetrix U95A gene arrays to map the effects of these factors on the HUVEC transcriptome. These experiments followed a paired design and were biologically replicated three times. In addition, one experiment was repeated using serial analysis of gene expression (SAGE). In contrast to some previous studies, we found that VEGF-A and PlGF consistently regulated only small, non-overlapping and culture media-dependant sets of HUVEC transcripts, despite causing significant cell biological changes.

Molecular classification of human endometrial cycle stages by transcriptional profiling.

Endometrium is a dynamic tissue that undergoes cyclic changes each month, under the overall control of estrogen and progesterone. The aims of this study were to investigate the changing global gene expression profile of human endometrium during the menstrual cycle using microarray technology and to determine the correlation between histopathological evaluation and molecular profile of the samples. Standard two-colour cDNA microarrays were performed on the 43 samples against a common reference, using a 10.5 K cDNA glass slide microarray. The results were validated using real-time PCR. Analysis of expression data was carried out using parametric analysis of variance with Benjamini-Hochberg correction. Hierarchical clustering reveals a strong relationship between histopathology and transcriptional profile of the samples. The study identified 1452 genes that showed significant changes in expression (P< or =0.05) across the menstrual cycle, with 425 genes having changes that are at least 2-fold. The data were also independently analysed by a CSIRO algorithm called GeneRaVE that identified a small subset of genes whose expression profiles could be used to classify nearly all the biopsies into their correct cycle stage. We also identified and validated three genes [(natural cytotoxicity triggering receptor (NCR)3, fucosyl transferase (FUT)4 and Fyn-binding protein (FYB)] that had not been shown to have significant cyclic changes in the human endometrium, previously. We have shown for the first time that endometrial cycle stage prediction is possible based on global gene expression profile.

Fibroids display an anti-angiogenic gene expression profile when compared with adjacent myometrium.

The aetiology of uterine fibroids remains unknown, despite causing significant gynaecological morbidity. Fibroids have a reduced microvascular density when compared with adjacent myometrial tissue. The aim of this study was to identify genes with differential expression between fibroid and adjacent normal myometrium, particularly genes with a role in angiogenesis. Total RNA was extracted from fibroid/myometrium pairs from 12 hysterectomy specimens, and used to perform 24 cDNA microarrays. There were 10,500 genes screened on each microarray for differential expression. Analysis of expression data was carried out using multiple t-tests, as well as a novel class prediction algorithm (GeneRaVE). The differential gene expression of selected genes was confirmed by quantitative 'real time' RT-PCR. Selected genes with a role in angiogenesis were further analysed for expression in isolated cell populations of endothelial cells (fibroid and myometrium) and smooth muscle cells (fibroid and myometrium), to see if their expression was confined to particular cell types. Twenty-five genes with differential gene expression between fibroid and myometrium were identified. Insulin-like growth factor-2, endothelin A receptor, connective tissue growth factor (CTGF), cysteine-rich angiogenic inducer 61 (CYR61) and collagen 4alpha2 (COL4A2) were confirmed by RT-PCR. CTGF and CYR61, both angiogenesis promoters, were reduced in expression relative to myometrium. COL4A2, the precursor for an angiogenesis inhibitor, canstatin, was increased relative to myometrium. These three genes display an anti-angiogenic expression profile in fibroids relative to myometrium. These findings may explain the reduced microvascular density seen in fibroids relative to myometrium.

Genomics in obstetrics and gynaecology.

With the Human Genome Project complete, and microarray technology progressing rapidly, the study of whole genomes has become a reality. The emerging field of genomics is full of promise, has become a cornerstone of commercial drug development, and looks certain to make a major contribution to clinical practice in the future. There is an increasing number of genomic studies concerned with obstetric and gynaecological conditions. Despite this, clinicians in their busy practices often lack a basic understanding of genomics and the tools involved in generating genome-based information. In the present review, we aim to provide the clinician with a basic overview of genomics--what it is, what tools it uses, and how it may benefit our patients. The existing published reports on genomic studies in the reproductive field is reviewed.

Migrant fathers and their attitudes to potential male hormonal contraceptives.

The purpose of this study was to assess potential uptake of male hormonal contraception (MHC) in migrant fathers in a post-partum setting, and to compare them to Australian-born fathers. It was a cross-sectional study of a convenience sample from the post-natal ward of a tertiary level obstetric hospital. Seventy-six English-speaking fathers born in South-East Asia or on the Indian subcontinent were surveyed and their responses compared with those provided by 118 Australian-born participants from a previous study. The main outcome measures were acceptability of potential male hormonal contraception on a 5-point scale, and preferred mode of administration. Information on past and future intended contraceptive use, including existing male forms of contraception, was also compared with data collected from the group of Australian-born men. Only 13.6% (95%CI: 5.8-21.4) of migrant fathers would definitely or probably consider trying MHC compared with 47.5% (95%CI: 38.5-56.5) of Australian-born fathers (chi-square, p < 0.001). There were significant differences in desired mode of administration in potential 'triers' from both groups, as well as in attitudes to existing contraception. Two-yearly injection was the most popular method of administration in migrants, with 38.3% of 'triers' listing it as their first choice (compared with 21.4% in published data on Australian-born men; chi-square, p = 0.038). We conclude that migrant groups are less enthusiastic about novel potential MHC. The influence of education on acceptance of this contraceptive possibility needs to be assessed.

Human cloning 2001.

This review summaries human cloning from a clinical perspective. Natural human clones, that is, monozygotic twins, are increasing in the general community. Iatrogenic human clones have been produced for decades in infertile couples given fertility treatment such as ovulation induction. A clear distinction must be made between therapeutic cloning using embryonic stem cells and reproductive cloning attempts. Unlike the early clinical years of in vitro fertilization, with cloning there is no animal model that is safe and dependable. Until there is such a model, 'Dolly'-style human cloning is medically unacceptable.