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The use of hemizona assay in the evaluation of the optimal sperm preparation technique.

The objective of the study was to evaluate the benefit of different sperm preparation methods by using the hemizona assay. A total of 58 men admitted to the male infertility clinic for evaluation were tested by routine semen analysis and hemizona assay. Five different techniques (swim-up, TEST-yolk buffer, Percoll, pentoxifylline and progesterone) were used for preparation of sperm suspensions. The effect of these treatments on the sperm-binding capacity using the hemizona assay was assessed. The routine swim-up preparation was used as the reference method. Of the four preparation methods, only the TEST-yolk buffer and pentoxifylline exhibited an overall statistically significant improvement in sperm-binding capacity in comparison with the swim-up preparation method (P = 0.01 and 0.001 respectively). Following preparation with Percoll and progesterone there was no change in the mean value of binding capacity, compared with swim-up. However, examination of the effect of the four treatments on each specimen individually yielded a diversity in the response, e.g. having the capability to enhance, damage or be ineffective in sperm binding capacity. The results support the conclusion that in-vitro sperm preparation methods can affect sperm binding to the zona pellucida. Since there is a diversity in the response of sperm samples to different treatments, the hemizona assay can be used in selecting the optimal sperm preparation method prior to its use for assisted reproductive techniques. This is advocated mainly for the 'male factor' group.

Cell Separation↗

Semen banking in patients with cancer: 20-year experience.

Modern techniques of banking sperm provide an effective way to preserve the option of future fertility for most teenagers and young men diagnosed with a variety of malignancies that will necessitate treatment with chemotherapy, pelvic surgery, or significant radiation doses to the testes. Results of cumulative data collected at the Cleveland Clinic Foundation from patients with testicular cancer, lymphoma, leukemia, sarcoma, carcinoma and other kinds of malignancy have revealed that: (1) pretreatment semen quality (pre-freeze and post-thaw) in patients with cancer is poorer compared with healthy donors; (2) the percentage decline in semen quality (from pre-freeze to post-thaw) in patients with cancer is similar to that of normal donors. This suggested that the effect of cryodamage on spermatozoa from patients with cancer is similar to that of normal donors. (3) The stage of cancer in patients with testicular cancer and Hodgkin's disease shows no relationship to their semen quality. Based on studies conducted at the Cleveland Clinic Foundation, we recommend that sperm cryopreservation be offered to all men of reproductive age who have malignancies. Cryopreservation is safe and inexpensive, and gives patients a chance to establish pregnancies in the future with an assisted reproductive technique.

Feasibility Studies↗

Excisional surgery versus ablative surgery for ovarian endometriomata: a Cochrane Review.

BACKGROUND: The objective of this review was to determine which is the most effective technique for treating an ovarian endometrioma; excision or ablation. METHODS: A systematic review employing the principles of the Cochrane Menstrual Disorders and Subfertility Group was undertaken. No randomized studies of the management of endometriomata by laparotomy were found. Two randomized studies of the laparoscopic management of ovarian endometriomata of >3 cm in size were included. RESULTS: Laparoscopic excision of the cyst wall of the endometrioma was associated with a reduced rate of recurrence of the endometrioma [odds ratio (OR) 0.41, confidence interval (CI) 0.18-0.93], reduced requirement for further surgery (OR 0.21, CI 0.05-0.79), reduced recurrence rate of the symptoms of dysmenorrhoea (OR 0.15, CI 0.06-0.38), dyspareunia (OR 0.08, CI 0.01-0.51) and non-menstrual pelvic pain (OR 0.10, CI 0.02-0.56). It was also associated with a subsequently increased rate of spontaneous pregnancy in women who had documented prior subfertility (OR 5.21, CI 2.04-13.29). CONCLUSIONS: There is some evidence that excisional surgery for endometriomata provides for a more favourable outcome than drainage and ablation, with regard to the recurrence of the endometrioma, recurrence of symptoms and subsequent spontaneous pregnancy in women who were previously subfertile. Consequently this should be the favoured surgical approach. However, we found no data to indicate the best surgical approach in women planning to undergo assisted reproductive techniques.

Endometriosis↗

Proteome analysis of human follicular fluid.

We used proteomic approach to analyze the protein profile of human follicular fluid (HFF) obtained from 25 normo-ovulatory women undergoing assisted reproduction techniques due to a male infertility factor. In all HFF samples analyzed we found 695 common spots distributed in the 3 to 10 pH range and in the 10-200 kDa range. Only 625 of these spots were also present in the plasma. We used MALDI-TOF-MS analysis to unequivocally assign 183 HFF/plasma matched spots and 27 HFF/plasma unmatched spots. A large number of acute-phase proteins, including transferrin, ceruloplasmin, afamin, hemopexin, haptoglobin and plasma amyloid protein, were identified in HFF in relatively high concentration supporting the hypothesis that mammalian ovulation can be compared to an inflammatory event. We also identified several important antioxidant enzymes; i.e., catalase, superoxide dismutase, glutathione transferase, paraoxonase, heat shock protein 27 and protein disulfide isomerase. This indicates that during maturation the human follicle is well protected against toxic injury due to oxidative stress.

Electrophoresis, Gel, Two-Dimensional↗

Sexual function in spinal cord lesioned men.

STUDY DESIGN: Review of literature. OBJECTIVE: To review the physical aspects related to penile erection, ejaculatory dysfunction, semen characteristics, and techniques for enhancement of fertility in spinal cord lesioned (SCL) men. SETTING: Worldwide: individuals with traumatic as well as non-traumatic SCL. RESULTS: Recommendations for management of erectile dysfunction in SCL men: If it is possible to obtain a satisfactory erection but of insufficient duration, then try to use a venous constrictor band to find out if this is sufficient to maintain the erection. Otherwise we recommend Sildenafil. If Sildenafil is not satisfactory then use intracavernous injection with prostaglandin E(1) (some SCL men may prefer cutaneous or intraurethral application). We discourage the implantation of penile prosthesis for the sole purpose of erection. Recommendations for management of ejaculatory dysfunction in SCL men: Penile vibratory stimulation (PVS) to induce ejaculation is recommended as first treatment choice. If PVS fails, SCL men should be referred for electroejaculation (EEJ). Semen characteristics: Impaired semen profiles with low motility rates are seen in the majority of SCL men. Recently reported data gives evidence of a decline in spermatogenesis and motility of ejaculated spermatozoa shortly after (few weeks) an acute SCL. It is suggested that some factors in the seminal plasma and/or disordered storage of spermatozoa in the seminal vesicles are mainly responsible for the impaired semen profiles in men with chronic SCL. Fertility: Home insemination with semen obtained by PVS and introduced intravaginally in order to achieve successful pregnancies may be an option for some SCL men and their partners. The majority of SCL men will further enhance their fertility potential when using either PVS or EEJ combined with assisted reproduction techniques such as intrauterine insemination or in vitro fertilization with or without intracytoplasmic sperm injection.

Alprostadil↗

Excisional surgery versus ablative surgery for ovarian endometriomata.

BACKGROUND: Endometriomata are endometriotic deposits within the ovary. The surgical management of these blood filled cysts is controversial. The laparoscopic approach to the management of endometriomata is favoured for as it offers the advantage of a shorter hospital stay, faster patient recovery and decreased hospital costs. Currently the commonest procedures for the treatment of ovarian endometriomata are either excision of the cyst capsule or drainage and electrocoagulation of the cyst wall. OBJECTIVES: The objective of this review was to determine the most effective technique of treating an ovarian endometrioma; either excision of the cyst capsule or drainage and electrocoagulation of the cyst wall, with regard to relief of pain, recurrence of the endometrioma, recurrence of symptoms and the subsequent spontaneous pregnancy rate. SEARCH STRATEGY: The reviewers searched the Cochrane Menstrual Disorders and Subfertility Group specialised register of trials (searched 15 Nov 2004), the Cochrane Register of Controlled Trials (The Cochrane Library, Issue 4, 2004), MEDLINE (1966-Nov 2004), EMBASE (1980- Nov 2004) and reference lists of articles, the handsearching of relevant journals and conference proceedings and by contacting leaders in the field of endoscopic surgery throughout the world. SELECTION CRITERIA: Randomised controlled trials of excision of the cyst capsule versus drainage and electrocoagulation of the cyst in the management of ovarian endometriomata. DATA COLLECTION AND ANALYSIS: Reviewers assessed eligibility and trial quality. MAIN RESULTS: No randomised studies of the management of endometriomata by laparotomy were found. Two randomised studies of the laparoscopic management of ovarian endometriomata of greater than 3cm in size were included. Laparoscopic excision of the cyst wall of the endometrioma was associated with a reduced rate of recurrence of the endometrioma (OR 0.41 CI 0.18-0.93), reduced requirement for further surgery (OR 0.21 CI 0.05-0.79), reduced recurrence rate of the symptoms of dysmenorrhoea (OR 0.15 CI 0.06-0.38), dyspareunia OR 0.08 CI 0.01-0.51) and non-menstrual pelvic pain (OR 0.10 CI 0.02-0.56). It was also associated with a subsequent increased rate of spontaneous pregnancy women who had documented prior sub-fertility (OR 5.21 CI 2.04-13.29). AUTHORS' CONCLUSIONS: There is some evidence that excisional surgery for endometriomata provides for a more favourable outcome than drainage and ablation, with regard to the recurrence of the endometrioma, recurrence of symptoms and subsequent spontaneous pregnancy in women who were previously subfertile. Consequently this approach should be the favoured surgical approach. However we found no data as to the effect of either approach in women who subsequently undergo assisted reproductive techniques.

Drainage↗

Assisted reproductive technology : experience with IVF/ICSI.

OBJECTIVE: To analyze success rates of assisted reproduction techniques ART (IVF/ICSI) in couples experiencing subfertility, and factors which may help in predicting treatment outcome. DESIGN: A prospective cohort follow-up study. PLACE AND DURATION OF STUDY: Study was conducted at Concept Fertility Centre, Karachi, between July 1, 2000 and December 31, 2002. SUBJECTS AND METHODS: A cohort of 310 couples undergoing 330 in-vitro fertilization (IVF) and intra cytoplasmic sperm injection (ICSI) cycles were followed for treatment outcomes. Predictor variables included age of women, cause of subfertility, drug regimen used for ovulation induction, day of embryo transfer, and ease of embryo transfer. While outcome variables included rates of fertilization, pregnancy, live birth, miscarriages, multiple pregnancies and ovarian hyperstimulation syndrome (OHSS). RESULTS: The overall pregnancy rate was 35.2 %, while takehome live-birth rate was 25.1%. Treatment cycles which were done for male factor only, had higher probability of success as compared to female factors or for unexplained infertility. Patients who underwent transvaginal ovum pick up and embryo transfer (TVOPU + ET) had an overall pregnancy rate of 29.4% as compared to those who underwent frozen embryo transfer (FET) cycles (22.6%). There was no statistically significant difference between pregnancy rates for ovulation induction with recumbent FSH (rFSH) and HMG (32.7% v 33.6%, p-value 0.87). Quality of embryo transfer was significantly better (33.7%, v 24.2%, p-value 0.042) in patients who conceived. CONCLUSION: Our results are comparable in terms of pregnancy rates, live birth rates and complication rates, to the results reported in international literature. We however, need more data from centres in Pakistan for future reference and research.

Adult↗

The aetiology and management of erectile, ejaculatory, and fertility problems in men with diabetes mellitus.

Erectile impotence is more common in the diabetic than the general population, occurs at a younger age, and is often associated with ejaculatory problems. For these, and possibly for other more subtle reasons, fertility may be a problem for men with diabetes. The symptoms of erectile and ejaculatory dysfunction are frequently not discussed between patient and doctor. Psychological factors are important but the vast majority of diabetic patients have an organic basis for their impotence. Both neurogenic and vascular factors are important in the pathogenesis of erectile failure. Autonomic neuropathy is almost certainly the cause of the ejaculatory failure that may be present in up to 40% of men with diabetes. The final biochemical mediator of erection within the penile erectile tissue is nitric oxide and a key enzyme in its degradation is phosphodiesterase (type V). Drugs that affect the metabolism of this enzyme are being developed to treat erectile failure. At present, the self injection of intra-cavernosal erectogenic agents (such as prostaglandin E1) provide the main form of therapy for erectile failure. Vacuum devices are a simple alternative and venous ligation surgery may be effective for a properly selected cohort of patients. Prosthetic implants are a final option for patients in whom all else has failed. Fertility problems, particularly when associated with ejaculatory failure can be overcome with modern assisted reproductive techniques. Nowadays, these will frequently involve gamete micro-manipulation.

Diabetes Complications↗

Early prenatal sonographic diagnosis of twin triploid gestation presenting with fetal hydrops and theca-lutein ovarian cysts.

The presence of theca-lutein ovarian cysts in the early second trimester of pregnancy is highly suspicious for a complete hydatidiform molar pregnancy but can be seen in association with a partial mole. Theca-lutein cysts may occur following hormonal stimulation for assisted reproductive techniques or in association with multiple gestations. Rare causes include immune and nonimmune fetal hydrops, maternal hypothyroidism, and triploid gestations. We report a case of a monochorionic twin gestation in which prenatal sonography demonstrated multiple anomalies and hydrops in each twin and bilateral theca-lutein ovarian cysts. Triploidy in both twins and a partial hydatidiform mole were confirmed at pathologic examination.

Abnormalities, Multiple↗

Embryo freezing for preventing Ovarian Hyperstimulation Syndrome.

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic potentially life threatening condition resulting from an excessive ovarian stimulation. Its reported incidence varies from 1% to 10% of in vitro fertilization (IVF) cycles. The factors leading to this syndrome have not been completely explained. It seems likely that the release of vasoactive substances, secreted by the ovaries under human chorionic gonadotropin (hCG) stimulation plays a key role in triggering this syndrome. The hallmark of this condition, is a massive shift of fluid from the intra-vascular compartment to the third space resulting in profound intra-vascular depletion and hemoconcentration. OBJECTIVES: To evaluate (i) the effectiveness of cryopreservation (embryo freezing) for the prevention of OHSS when compared with human intra-venous albumin infusion and (ii) the effectiveness of the elective cryopreservation (embryo freezing ) of all embryos for the prevention of OHSS when compared with fresh embryo transfer. SEARCH STRATEGY: Publications in the literature that describe or may describe randomised controlled trials of both human intra-venous albumin and freezing of all embryos in the management of OHSS as consequence of the superovulation in assisted reproduction techniques (ART) cycles were searched. The Cochrane Menstrual Disorders and Subfertility Review Group specialised register of controlled trials was searched. In addition, MEDLINE (PUBMED 1985 to 2001), EMBASE (1985 to 2001), CINHAL (1985 to 2001) and the National Research Register were searched SELECTION CRITERIA: Randomised controlled trials (RCTs) in which either human intra-venous albumin or cryopreservation of all embryos were used as a therapeutic approach to OHSS were included. The participants were women of reproductive age who were down regulated by gonadotrophin-releasing hormone-analogue (GnRH-a), undergoing superovulation in in vitro fertilization/intra-cytoplasmic sperm injection (IVF/ICSI) cycles. DATA COLLECTION AND ANALYSIS: Two reviewers, Mr N.N. Amso (NNA) and Dr A. D'Angelo (ADA) scanned the titles and the abstracts of the reports identified by electronic searching in order to find relevant papers. One reviewer (ADA) obtained copies of the full text articles and made copies for the other reviewer (NNA) in which details of authors, institution, results and discussion were removed in order to assess their eligibility for inclusion. Then, both reviewers extracted data independently using forms designed according to Cochrane guidelines. Disagreements were resolved by discussion. Additional information on the trial methodology or data were requested by writing to the corresponding authors directly. The interventions compared were cryopreservation (embryo freezing) versus intra-venous human albumin administration and elective cryopreservation of all embryos versus fresh embryo transfer. The primary outcomes were: incidence of moderate and severe OHSS versus nil/mild OHSS, clinical pregnancies/woman. The secondary outcomes were: number of oocytes retrieved, number of oocytes fertilized, number of embryos transferred, number of embryos frozen, multiple pregnancy rate, live birth rate, number of women admitted to the hospital as inpatient or outpatient and time to the next menstrual period (resolution time). Statistical analysis was performed in accordance with the Cochrane Menstrual Disorders and Subfertility Group guidelines. MAIN RESULTS: Seventeen studies were identified, two of which met our inclusion criteria. One study was included where cryopreservation (embryo freezing) was compared with intra-venous human albumin administration (Shaker 1996) and one study was included where elective cryopreservation of all embryos was compared with fresh embryo transfer (Ferraretti 1999). When cryopreservation was compared with intra-venous human albumin administration no difference was found in all the outcomes examined between the two groups. When elective cryopreservation of all embryos was compared with fresh embryo transfer no difference was found in all the outcomes examined between the two groups. REVIEWER'S CONCLUSIONS: This review has showed that there is insufficient evidence to support routine cryopreservation and insufficient evidence for the relative merits of intra-venous albumin versus cryopreservation.

Albumins↗

Gonadotrophins for idiopathic male factor subfertility.

BACKGROUND: Male factor infertility accounts for 50% of infertility. The treatment of idiopathic male infertility is empirical. Urinary, purified, and recombinant gonadotrophins have been used to improve sperm parameters in idiopathic male infertility with the goal of increasing pregnancy rates. Research addressing pregnancy rates in partners of men treated with gonadotrophins has had conflicting results and needs to be analysed. OBJECTIVES: To determine the effectiveness of gonadotrophin administration to men with idiopathic subfertility on spontaneous pregnancy rate and in assisted reproductive techniques (ARTs). SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders and Subfertility Group trials register (19 November 2004), the Cochrane Central Register of Controlled Trials (The Cochrane Library, issue 4, 2004), MEDLINE (1966 to April 2005), EMBASE and Biological Abstracts (1980 to November 2004). Searches were not limited by language. The bibliographies of included, excluded trials and abstracts of major meetings were searched for additional trials. Authors and pharmaceutical companies were contacted for missing and unpublished data. SELECTION CRITERIA: Truly randomised controlled trials where gonadotrophins were administered for the treatment of idiopathic male subfertility with reporting of pregnancy rates were included in the review. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Study authors were contacted for additional information. Adverse effects information was collected from the trials. We analysed data regarding pregnancy occurring within 3 months after gonadotrophin therapy. MAIN RESULTS: Four RCTs with 278 participant were included in the analysis. None of the studies had an adequate sample size and they had variable follow-up periods. None of the studies reported live birth or miscarriage rates. Compared to placebo or no treatment, gonadotrophins showed a significantly higher pregnancy rate per couple randomized within 3 months of completing therapy (OR 3.03, 95% CI 1.30 to 7.09). Pregnancy rate was 13.4% (19/142) in the gonadotrophin group and 4.4% (6/136) in the control group. AUTHORS' CONCLUSIONS: The number of trials and participants is insufficient to draw final conclusions. A large multicenter study with adequate power is needed.

Follicle Stimulating Hormone↗

A comparison of stated preference methods for estimating monetary values.

This paper presents a comparison of willingness to pay (WTP) estimates generated from a dichotomous choice (DC) contingent valuation experiment and a choice experiment (CE). The study was conducted with subjects undergoing assisted reproductive techniques in the Grampian area of Scotland. Subjects first completed a DC WTP questionnaire. They were sent a CE questionnaire two months later. Welfare estimates for the DC WTP study and CE were not significantly different. Important research questions are raised concerning the validity of the various approaches to estimating WTP.

Financing, Personal↗

The use of novel semen quality scores to predict pregnancy in couples with male-factor infertility undergoing intrauterine insemination.

The purpose of this study was to determine whether 2 new semen quality (SQ) scores could predict pregnancy in patients undergoing intrauterine insemination (IUI) for male-factor infertility and whether an overall score could help in counseling these couples with assisted reproductive technique (ART) options. Ninety-three couples with male-factor infertility were examined for semen analysis. Samples were prepared by density gradient separation (47% and 90%), and IUI was performed. On the basis of the 2 semen scores (SQ and relative quality [RQ]), the IUI-semen pregnancy score (IUI-SPS) was calculated. Of the 192 IUI cycles, 14% (27 of 192) resulted in pregnancy. Both prewash SQ and RQ scores were significantly related to pregnancy (P =.02 and P <.001), as was the postwash RQ score (P <.001). Of the IUI cycles in which the postwash RQ score was greater than 125, 40% (13 of 32) resulted in pregnancy compared to 9% of cycles (14 of 160) in which the postwash SQ score was less than 125. The prewash IUI-SPS score was significantly related to IUI-induced pregnancy (P <.001). Both the pre- and postwash SQ and RQ scores can predict pregnancy in male infertility patients undergoing IUI. Patients with an IUI-SPS less than 150 may be advised to seek in vitro fertilization (IVF), whereas those with an IUI-SPS greater than 150 may be advised to seek IUI.

Female↗

Concentration of glutathione and expression of glutathione peroxidases 1 and 4 in fresh sperm provide a forecast of the outcome of cryopreservation of human spermatozoa.

Oxidative stress imbalance potentially leads to damage of the structure of the cell and macromolecules such as plasma membrane components, proteins, and DNA. The plasma membrane of the sperm cell, which has high levels of polyunsaturated fatty acids, renders it particularly sensitive to free radical-mediated attacks. The freezing and subsequent thawing of sperm is a physically stressful process carried out during routine procedures in assisted reproduction techniques, which results in a highly variable and unpredictable reduction in the number of motile sperm cells. Subsequently, oxidative status can positively or negatively affect the motility, viability, and fertilizing capacity of thawed sperm. These effects are counteracted by various oxidative defense enzymes and anti-oxidants such as glutathione peroxidase isoforms GPx1 and GPx4, glutathione reductase (GR), and cellular glutathione (reduced) (GSH). In this way, oxidative status could represent a predictive marker of sperm quality following the freeze-thaw process. This study was based on 56 human sperm samples. We observed direct positive and negative relationships between the postthaw motile sperm recovery rate and GPx1 and GPx4 expression and activity, on the one hand, and GSH concentrations, on the other. No correlation was found between this recovery rate and GR or basic semen parameters. Predictive values clearly demonstrate that, among the molecules analyzed, the most accurate diagnoses result when analyses are conducted for GPx1 and GPx1 messenger RNA expression, GPx1 and GPx4 enzymatic activity, and GSH concentration. In conclusion, a reserve of glutathione, together with GPx expression, is necessary to eliminate free radicals using GSH or a like structural protein and seems to be essential for a good postthaw recovery. These molecules can be employed as indicators of postthaw sperm quality.

Cryopreservation↗

Sperm cryopreservation for men with nonmalignant, systemic diseases: a descriptive study.

Cytotoxic drugs and immunosuppressive therapies are used to treat patients with nonmalignant, nontesticular systemic diseases. These therapies can permanently suppress spermatogenesis. Sperm cryopreservation before treatment theoretically could give these men the opportunity to achieve a pregnancy with a woman later in life when the couple decides to do so. However, it is not known whether pretreatment sperm quality in these men is good enough to be used for assisted reproductive techniques. The main objective of this study was to determine the usefulness of cryopreservation in this patient population by: 1) assessing their pretreatment semen quality (eg, count, motility, and motion kinetics) and comparing it with that of healthy donors before and after cryopreservation; 2) comparing patients' pretreatment semen characteristics with World Health Organization reference values for normal sperm; and 3) examining the differences in semen parameters among patient groups. Semen specimens were obtained from 25 healthy donors and from 23 patients with a variety of disorders (12 had autoimmune disorders, 4 had kidney disorders, 3 had diabetes, 2 had ulcerative colitis, and 2 had heart transplants). All patients, except those with diabetes, required immunosuppressive or cytotoxic therapy. Although the pretreatment quality of the semen of these patients was not as good as that of donors, semen samples were within the normal reference range of the World Health Organization. No statistically significant differences in sperm parameters were found within the 4 patient groups except for those with diabetes (n = 3), who showed poorer sperm counts (P < .04). However, no conclusive evidence can be reached due to the small sample size. Our results indicate that pretreatment semen quality in these patients is adequate for reproductive techniques. We believe that cryopreservation should be offered to patients of reproductive age with disease or treatment regimens that may cause infertility.

Adult↗

Cervical length, multifetal pregnancy reduction, and prediction of preterm birth.

PURPOSE: To evaluate the application of transvaginal sonography assessment of cervical length before fetal reduction for predicting spontaneous preterm birth in triplet gestations reduced to twins. METHODS: This retrospective study was conducted at the ultrasound unit of a university-affiliated municipal hospital. The study cohort consisted of 25 women with triplet gestations following ovulation induction or assisted-reproduction techniques who underwent fetal reduction to twins. Cervical length was assessed via transvaginal sonography before fetal reduction, and data on pregnancy outcome were retrieved from maternal records and/or maternal interviews. RESULTS: Cervical length (mean +/- SD) at reduction was 4.0 +/- 0.85 (range: 1.2-5.5). Five women were excluded from statistical evaluation because pregnancy complications precluded spontaneous delivery. Two of 3 (67%) women with a cervical length of <3.5 cm delivered prior to 33 weeks' gestation compared with 1/17 (6%) women with a cervical length > or = 3.5 cm. This difference was statistically significant (P < 0.05). The sensitivity, specificity, positive predictive value, and negative predictive value of cervical lengths <3.5 cm to predict delivery prior to 33 gestational weeks was 67%, 94%, 67%, and 94%, respectively. CONCLUSIONS: Measurement of cervical length in triplet pregnancies before fetal reduction provides useful predictive information on the risk for preterm delivery.

Adult↗

Ultrastructure of oocyte maturation, fertilization, and early embryo development in vitro in the Siberian tiger (Panthera tigris altaica).

The application of assisted reproduction techniques to wild cats has been stalled by a lack of basic knowledge of the reproductive biology in these species. In this study, the ultrastructure of Siberian tiger (Panthera tigris altaica) cumulus-oocyte-complexes (COCs), as well as in vitro produced (IVP) zygotes and embryos were investigated, to estimate the normality of the manipulated reproduction processes. Adult female tigers were subjected to a purified porcine pFSH/pLH stimulation treatment followed by oocyte aspiration. According to morphological appearance at the stereomicroscopical level, COCs were classified as mature, immature, or degenerated, and then allocated into the following groups: presumptively immature COCs, which were in vitro matured (IVM-group) before fixation; presumptively mature COCs, which were either fixed after retrieval (pre-IVF-group), following in vitro insemination (IVF-group) or following in vitro insemination and subsequent in vitro culture (IVC-group). All specimens were processed for light and transmission electron microscopy (TEM). Both the IVM- and pre-IVF-group included oocytes in meiotic stages ranging from prophase I to metaphase II, and some prophase I oocytes in the IVM-group were apparently in their growth phase. The IVF-group presented features of presumptive normal fertilization, but aberrations such as polynucleation were also noted. The IVC-group included cleavage stage embryos of which, however, many were polynucleated. In conclusion, the procedures used for stimulation, aspiration, and classification of COCs resulted in retrieval of a heterogeneous population of oocytes which, following IVF and IVC, displayed a high rate of developmental deviations.

Animals↗

Active demethylation of individual genes in intracytoplasmic sperm injection rabbit embryos.

Intracytoplasmic sperm injection (ICSI), as an assisted reproduction technique, has been widely used in animal and human. However, its possible effect on epigenetic changes has not been well studied. To investigate whether ICSI can induce aberrant DNA methylation changes in rabbit preimplantation embryos, we examined the methylation status of the SP-A promoter region and the satellite sequence Rsat IIE by bisulfite-sequencing technology. The SP-A promoter region was extensively demethylated before the first round of DNA replication commences, and the unmethylated status was maintained until morula when dynamic remethylation occurred. A similar but more moderate demethylation process was observed in satellite sequence Rsat IIE. These results are in contrast with the previous reports of no active demethylation in normal rabbit embryos, suggesting that the active demethylation we observed may be induced by ICSI.

Animals↗