MedWorm: Reproduction Medicine

Consensus on infertility treatment related to polycystic ovary syndrome

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

(Source: Human Reproduction)

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Reply: ethical recruitment of patients for pgs trial

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

(Source: Human Reproduction)

Ethical recruitment of patients for pgs trial

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

(Source: Human Reproduction)

Genetic polymorphisms of esr1 and esr2 that may influence estrogen activity and the risk of hypospadias

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The etiology of hypospadias is regarded as a complex disorder with both genetic and environmental contributions. Although alterations in androgen activity have been associated with hypospadias, few associations with estrogen activity have been documented. Here, we assessed genetic polymorphisms in estrogen receptor genes and their association with hypospadias. METHODS Using a case–control study of 59 cases with hypospadias and 286 controls, we examined the association of hypospadias with the following polymorphisms: PvuII and XbaI in ESR1, and 2681-4A>G in ESR2. RESULTS For the cases, we found a negative association with the G allele containing variants of ESR1 XbaI (OR = 0.52, P < 0.05), and a negative association with the G allele containing variants of ESR2 2681-4A>G (OR = 0.59, P < 0.05). For the cases, we also identified a negative association with the CG haplotype, and a positive association with the CA haplotype, defined by ESR1 PvuII and XbaI (P < 0.05). CONCLUSIONS These findings suggest that the G allele containing variants of ESR1 XbaI and the G allele containing variants of ESR2 2681-4A>G may decrease the risk of hypospadias, whereas the ESR1 C-A haplotype may increase its risk. (Source: Human Reproduction)

Parathyroid hormone-responsive b1 gene is associated with premature ovarian failure

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Premature ovarian failure (POF) is a complex and heterogeneous disorder that is influenced by multiple genetic components. Here, we performed a two-stage association study to identify POF-associated genes. METHODS A first stage linkage disequilibrium (LD)-based genome-wide association study was performed using 24 pairs of patients with POF and matched controls and a high-throughput BeadChip assay with 109365 single-nucleotide polymorphisms (SNPs) that are scattered throughout the genome in an exon-centric and evenly spaced manner. A region that was shown to be strongly associated with POF was then tested again for POF association in the second stage by using a larger sample size (101 cases and 87 controls) and additional putative causal SNPs. RESULTS The first stage analysis revealed that many regions were associated with POF, with part of chromosome 7p14 that contains the parathyroid hormone responsive-B1 (PTHB1) gene showing the strongest association. A POF-susceptible haplotype of PTHB1 (ht1, ‘GAAAG’, P = 0.00034) and a POF-resistant haplotype (ht2, ‘TGTGC’) were also identified. The association between POF and two PTHB1 SNPs (rs3884597 and rs6944723) and part of ht1 was confirmed in the second stage analysis. The additional SNP, rs11773504, was considered as a putative causal variant causing an amino acid change, Ala to Thr. CONCLUSIONS We showed for the first time that PTHB1 is strongly associated with POF, and ht1 confers susceptibility to POF. While causative SNPs were not identified, the polymorphism of the non-synonymous SNP rs11773504 and the repeated association of ht1 with POF suggest that PTHB1 may contribute to POF pathogenesis. (Source: Human Reproduction)

A distinct cohort of the tgf{beta} superfamily members expressed in human endometrium regulate decidualization

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Successful blastocyst implantation requires the differentiation of human endometrial stromal cells (HESC), a process known as decidualization. Activin A, a transforming growth factor β (TGFβ) superfamily member, enhances HESC decidualization and localizes to decidual cells in human endometrium. Other TGFβ superfamily members, including BMP2, BMP4, BMP7, GDF5, GDF8, GDF11, TGFβs and Nodal, may also play a role during decidualization. This study aimed to identify these TGFβ family members in human endometrium, and to determine whether they are involved in human decidualization. METHODS Protein localization of TGFβ family members was examined in secretory phase human endometrium and first trimester decidua by immunohistochemistry. mRNA expression was examined in HESC. Activin inhibitors (Activin-M108A/SB431542) with differing specificities for the other TGFβ members under consideration were applied during HESC decidualization in vitro. The secretion levels of potential TGFβ superfamily members were measured during decidualization, and recombinant proteins added to examine their effect. RESULTS This study has identified BMP2, BMP4, BMP7, GDF5, GDF8 and GDF11 but not Nodal in secretory phase human endometrium, but only BMP2, GDF5 and TGFβ1 protein were detected in decidual cells. All ligands except Nodal were expressed by cultured HESC. Both inhibitors significantly reduced decidualization validating the role of activin, but potentially also other TGFβ members, during decidualization. BMP2 and TGFβ1 secretion increased during HESC decidualisation and exogenous administration of these proteins significantly enhanced decidualization in vitro. CONCLUSIONS Like activin, BMP2 and TGFβ1 are likely to be involved in HESC decidualization. This is the first study to identify and localize BMP4, BMP7, GDF5, GDF8 and GDF11 in secretory phase human endometrium. Understanding the factors critical for the implantation process is needed for improving fertility and pregnancy outcomes. (Source: Human Reproduction)

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Greek hyperinsulinemic women, with or without polycystic ovary syndrome, display altered inositols metabolism

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND We have shown that American women with polycystic ovary syndrome (PCOS) have decreased glucose-stimulated release of a putative mediator of insulin action, D-chiro-inositol (DCI)-containing inositolphosphoglycan (DCI-IPG), and increased urinary clearance of DCI (uClDCI), which was associated with hyperinsulinemia. METHODS DCI levels and the release of insulin and DCI-IPG during an oral glucose tolerance test (AUCs) were assessed in 27 Greek PCOS and 10 normal Greek women. RESULTS PCOS women were heavier than controls (BMI = 28.4 versus 23.7 kg/m2, P = 0.05) with higher waist-to-hip ratios (WHR = 0.78 versus 0.71, P = 0.009) and increased free testosterone (P = 0.048) and AUCinsulin (P = 0.04). In PCOS women, incremental AUCDCI-IPG was significantly decreased by 59% (2158 versus 5276%·min, P = 0.01), even after correction for BMI and WHR. Finally, increased uClDCI (r = 0.35, P = 0.04) and decreased AUCDCI-IPG (r = 0.46, P = 0.004) were significantly associated with hyperinsulinemia in all women together, even after correction for BMI and WHR (Ps = 0.02 and 0.007), and regardless of PCOS status. CONCLUSIONS Greek women, with or without PCOS, display increased uClDCI and decreased AUCDCI-IPG in association with higher insulin levels but independent of adiposity. Increased clearance of inositols might reduce tissue availability of DCI and decrease the release of DCI-IPG mediator, which could contribute to insulin resistance and compensatory hyperinsulinemia in Greek women, as previously described in American women. (Source: Human Reproduction)

Cardiovascular risks and metabolic syndrome in hong kong chinese women with polycystic ovary syndrome

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Women with polycystic ovary syndrome (PCOS) frequently exhibit central obesity, glucose intolerance, atherogenic dyslipidaemia and hypertension which are characteristic features of the metabolic syndrome (MetS). METHODS A total of 295 premenopausal Chinese women with PCOS diagnosed by the Rotterdam criteria (mean age: 30.2 ± 6.4 years) and 98 control subjects without PCOS were evaluated for prevalence of MetS and cardiovascular risk factors, including dyslipidaemia and dysglycaemia. RESULTS Using the 2005 modified Adult Treatment Panel III criteria, MetS (presence of three or more risk factors) was found in 24.9% of PCOS women compared to 3.1% of controls. The prevalence of MetS in PCOS women increased from 16.7% at under 30 years of age to 53.3% at over 40 years. MetS was also more prevalent in overweight and obese (41.3%) than normal-weight PCOS women (0.9%). However, multivariate regression analysis showed that women with PCOS had a 5-fold increase in risk of MetS (odds ratio 4.90; 95% confidence interval: 1.35–17.84) compared with women without PCOS even after controlling for age and BMI, suggesting PCOS alone is an independent risk factor for MetS. CONCLUSIONS There is high prevalence of MetS in Hong Kong Chinese women with PCOS despite their relatively young age. Recognition of these cardiometabolic risk factors requires a high level of awareness in conjunction with early and regular screening. (Source: Human Reproduction)

Predicting the fsh threshold dose in women with who group ii anovulatory infertility failing to ovulate or conceive on clomiphene citrate

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The objective of this investigation was to establish independent predictors of follicle-stimulating hormone (FSH) threshold dose in anovulatory women undergoing ovulation induction with FSH preparations. METHODS One hundred and fifty-one patients with WHO Group II anovulatory infertility failing to ovulate or conceive on clomiphene citrate underwent ovarian stimulation with FSH-only preparations following a low-dose step-up protocol. The individual FSH threshold dose was defined as the FSH dose when meeting the human chorionic gonadotrophin criteria (one follicle ≥17 mm, or 2–3 follicles ≥15 mm). The influence of demographics, physical characteristics, obstetric and infertility and menstrual cycle history, ovarian ultrasonography, endocrine parameters and type of gonadotrophin preparation on the FSH threshold dose was assessed through multiple regression analysis. RESULTS In the univariate analysis, age, body mass index (BMI), failure to ovulate with clomiphene citrate, menstrual cycle history (amenorrhea, oligomenorrhea or anovulatory cycles of 21–35 days), mean ovarian volume, LH/FSH ratio, testosterone and free androgen index were significant (P < 0.05) predictors of FSH threshold dose. In the multivariate analysis, menstrual cycle history, mean ovarian volume and BMI remained significant (P < 0.001). CONCLUSIONS The individual FSH threshold dose for ovulation induction in anovulatory women can be predicted based on three variables easily determined in clinical practice: menstrual cycle history, mean ovarian volume and BMI. A FSH dosage nomogram was constructed based on these parameters. (Source: Human Reproduction)

The endocrine and follicular growth dynamics throughout the menstrual cycle in women with consistently or variably elevated early follicular phase fsh compared with controls

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Elevated early follicular phase (EFP) FSH is frequently observed in subfertile patients. In these women, temporary normalization of FSH concentrations is known to occur. We studied the complete endocrine cycle profile of subfertile young women with elevated basal FSH compared with controls. METHODS Daily bloodsampling and ultrasound monitoring in the follicular phase was performed in 22 patients with elevated basal FSH levels (identified in screening) and in 16 controls during one menstrual cycle and for 5 days of the next cycle. RESULTS Eleven patients showed elevated basal FSH levels in the study cycle (‘High, High’; H,H group) whereas 11 had normalized basal FSH levels (‘High, Low’; H,L group). Anti-Müllerian hormone (AMH) was lower in both groups. In the H,H group, FSH was higher in all phases of the cycle and both inhibin A and B were lower during the EFP. In the H,L group, FSH was also higher than in controls in the EFP and the late luteal phase and inhibin A was higher in the periovulatory phase. ‘Normalization’ of Day 3 FSH in women with previously elevated FSH was associated with normalization of inhibin B levels in the preceding luteal phase. CONCLUSIONS The endocrine cycle profile in younger subfertile patients with consistently elevated basal FSH resembles that in published data from older women and also reflects a low ovarian reserve. Normalization of FSH in association with normal inhibin B suggests a temporary increase of the available cohort. (Source: Human Reproduction)

Hypoxia is responsible for soluble vascular endothelial growth factor receptor-1 (vegfr-1) but not for soluble endoglin induction in villous trophoblast

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Pre-eclampsia is a pregnancy disorder characterized by a maternal endothelial cell dysfunction associated with low levels of circulating placental growth factor (PlGF) and increased levels of total vascular endothelial growth factor (VEGF), soluble VEGF receptor-1 (sVEGFR-1), and soluble endoglin, a transforming growth factor β1 and 3 coreceptor. Here, we tested the hypothesis that these altered levels of angiogenic cytokines and of the anti-angiogenic soluble forms of cytokine receptors could be the consequence of hypoxia. METHODS Normal human umbilical vein endothelial cells, immortalized first trimester extravillous trophoblast cells (HTR8/SVneo) and first trimester placental villi explants (8–14 weeks) were used for culture under normoxia (20% O2) or hypoxia (1% O2). Culture media were collected for the measurement of cytokines by enzyme-linked immunosorbent assay. Total RNA was extracted for RT-PCR analysis. RESULTS Under hypoxia, villous trophoblast expressed higher levels of VEGF, VEGFR-1, sVEGFR-1 and VEGFR-2 mRNAs (P < 0.001), and secreted more VEGF and sVEGFR-1 proteins (P < 0.05). In contrast, PlGF mRNA and protein were decreased in 1% O2 (P < 0.001), whereas endoglin (Eng) was not modulated. Additionally, sVEGFR-1 directly abolished VEGF/PlGF-induced angiogenesis in the rat aortic ring assay. CONCLUSIONS Our results support the hypotheses that, in pre-eclampsia, (i) overproduction of VEGF family factors by pre-eclamptic placenta is a consequence of induced hypoxia; (ii) overproduction of sVEGFR-1 by hypoxic villous trophoblast accounts for maternal free VEGF depletion; (iii) low circulating level of free PlGF is not only related to sVEGFR-1 overproduction, but also to hypoxia-induced mRNA down-regulation; (iv) Eng is not modulated by hypoxia. (Source: Human Reproduction)

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Myeloid ecotropic viral integration site 1 (meis) 1 involvement in embryonic implantation

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The HOXA10 homeobox gene controls embryonic uterine development and adult endometrial receptivity. The three-amino-acid loop extension (TALE) family homeobox genes like myeloid ecotropic viral integration site 1 (MEIS) provide enhanced target gene activation and specificity in HOX-regulated cellular processes by acting as HOX cofactors. METHODS AND RESULTS Analysis of an Affymetrix data set in the public domain showed high expression of MEIS1 in human endometrium. MEIS1 expression was confirmed during the human menstrual cycle by RT–PCR and in situ hybridization and was increased during the secretory compared with proliferative phase of the cycle (P = 0.0001), the time of implantation. To assess the importance of maternal Meis1 expression in a mouse model, the uteri of Day 2 pregnant mice were injected with Meis1 over-expression or small interfering RNA (siRNA) constructs. Blocking Meis1 expression by siRNA before implantation significantly reduced average implantation rates (P = 0.00001). Increased or decreased Meis1 expression significantly increased or decreased the expression of integrin β3, a transcriptional target of HOXA10 and an important factor in early embryo-endometrium interactions (P = 0.006). Manipulating Meis1 expression before implantation also dramatically affected the number of pinopodes, uterine endometrial epithelial projections that develop at the time of endometrial receptivity. CONCLUSIONS The results suggest that in mouse, meis1 contributes to regulating endometrial development during the menstrual cycle and establishing the conditions necessary for implantation. (Source: Human Reproduction)

Native human zona pellucida glycoproteins: purification and binding properties

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Fertilization starts with the binding of the spermatozoa to the zona pellucida (ZP) of the oocyte. Such binding is a carbohydrate-mediated event and consists of a series of tightly regulated events. Molecular interactions between spermatozoon and ZP in human are not well characterized due to limited availability of oocytes for research. Our current technology cannot generate recombinant human ZP (hZP) glycoproteins with native glycosylation. METHODS AND RESULTS In this study, hZP glycoproteins, hZP2 (~120 kDa), hZP3 (~58 kDa) and hZP4 (~65 kDa) were purified from ZP (purity >88%) by immunoaffinity columns. The binding sites of the purified native hZP3 and hZP4 were localized to the acrosome region of the capacitated human spermatozoa, and were lost after acrosome reaction. Purified human hZP2 bound to this region only in acrosome-reacted spermatozoa. Differential binding of the three glycoproteins to the post-acrosomal region and the midpiece of the spermatozoa was observed. In addition, hZP3, but not hZP2 and hZP4, induced hyperactivation. The stimulatory activity was dependent partly on N-linked glycosylation of hZP3. CONCLUSIONS This manuscript describes the biological activities of purified hZP glycoproteins from the native source for the first time. (Source: Human Reproduction)

Contribution of the oocyte nucleus and cytoplasm to the determination of meiotic and developmental competence in mice

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Mammals have two types of full-grown oocytes: those with germinal vesicles (GVs) in which the chromatin is condensed and surrounds the nucleolus (surrounded-nucleolus (SN)-type) and those in which the chromatin is less condensed and does not surround the nucleolus (non-surrounded-nucleolus (NSN)-type). Although SN oocytes possess higher meiotic and developmental competence than NSN oocytes, the factors underlying this difference are unknown. METHODS AND RESULTS The GVs of murine SN and NSN oocytes were exchanged by nuclear transfer and the nucleus/cytoplasm of each reconstructed oocyte was classified as follows: SN/SN, NSN/SN, SN/NSN or NSN/NSN. After reconstruction, the meiotic maturation and preimplantation development of the oocytes were analysed. Few mature SN/NSN and NSN/NSN oocytes were observed (20–26%). In contrast, 88% of the NSN/SN oocytes matured; however, they rarely developed to the blastocyst stage after fertilization (4%), whereas most of the SN/SN oocytes matured (84%) and reached the blastocyst stage (83%). When the metaphase II (MII) plates of in vitro-matured NSN/SN oocytes were transferred into enucleated MII oocytes in which the contents of the SN-type GVs were spread into the cytoplasm, they completed full-term development. CONCLUSIONS The differences in meiotic and developmental competence between SN and NSN oocytes are determined by factors in the cytoplasm and nucleus, respectively. In addition, material(s) within SN-type GVs, and not the chromatin configuration itself, is essential for full-term development. (Source: Human Reproduction)

Changing etiology of tubal pregnancy following ivf

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Tubal pregnancy (TP) is twice as common following IVF when compared with natural conception. This is surprising, since embryo transfer is aimed for an accurate area in the uterine cavity. We thus hypothesized that either the embryo or the Fallopian tube actively participates in a pathological process leading to implantation outside the uterine cavity. Since we recently found that E-cadherin expression is a useful marker of endometrial receptivity, we considered that it may have a role in TP following IVF. Therefore, the aim of this study was to compare E-cadherin expression and localization in tubal implantation sites from spontaneous TP and TP post-IVF. METHODS We compared E-cadherin immunohistochemistry levels on cross-sections of Fallopian tubes in 11 spontaneous (antegrade) versus 13 post-IVF (retrograde) TP. The intensity of immunoreactivity was scored in a semi-qualitative blinded manner. RESULTS The semi-quantitative intensity score in IVF tubal samples was more than double that observed in spontaneous TP (16.9 versus 7.3, respectively, P < 0.0005). E-cadherin showed the most intense immunostaining in cytotrophoblast cells of chorionic villi in ectopic TP post-IVF compared with negative or weak staining in spontaneous ectopic TP. CONCLUSIONS E-cadherin can serve as a marker of implantation. Differential expression of this adhesion molecule in TP post-IVF, when compared with natural conception, may reflect a different mechanism of embryo implantation. Moreover, the observation that E-cadherin is mostly expressed in trophoblasts, and not in the tubal wall, suggests that the preimplantation embryo may actively participate in locating a suitable implantation site. (Source: Human Reproduction)

Risk factors associated with pregnancies containing a monochorionic pair following assisted reproductive technologies

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Although several factors have been identified to predispose to an increased incidence of monozygotic twinning in assisted reproductive technologies (ART), the relative risks associated with each have yet to be fully established. Moreover, the focus has been predominantly on monozygosity, which, in the absence of monochorionicity, does not increase perinatal risk. The present objective was to undertake an analysis of the relative risks of factors associated with monochorionic pairs resulting from ART. METHODS Study cycles included the last cycle, of each patient undergoing ART at Brigham and Women's Hospital from January 1998 to December 2004, that resulted either in a pregnancy with a monochorionic pair (n = 41) or a pregnancy without a monochorionic pair at 12 weeks (n = 2460). We used multivariable logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) to identify factors significantly associated with a monochorionic pair. RESULTS Independent predictors of a monochorionic pair were assisted hatching (OR 2.23, 95% CI 1.06–4.67), ICSI (OR 2.42, 95% CI 1.22–4.83) and Day 5 embryo transfer (OR 2.48, 95% CI 1.62–3.80). The effects of ICSI and Day 5 transfer were amplified when cycles involved both interventions. CONCLUSIONS ICSI and Day 5 embryo transfer synergistically increase the risk of monochorionic placentation. Patients undergoing these procedures should be counselled regarding these increased risks. (Source: Human Reproduction)

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Relevance of anti-mullerian hormone measurement in a routine ivf program

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Diminished ovarian reserve has become a major cause of infertility. Anti-Mullerian hormone (AMH) seems to be a promising candidate to assess ovarian reserve and predict the response to controlled ovarian hyperstimulation (COH). This prospective study was conducted to evaluate the relevance of AMH in a routine IVF program. METHODS Three hundred and sixteen patients were prospectively enrolled to enter their first IVF/ICSI-cycle. Age, FSH-, inhibin B- and AMH-levels and their predictive values for ovarian response and clinical pregnancy rate were compared by discriminant analyses. RESULTS A total of 132 oocyte retrievals were performed. A calculated cut-off level ≤1.26 ng/ml AMH alone detected poor responders (≤4 oocytes) with a sensitivity of 97%, and there was a 98% correct prediction of normal response in COH if levels were above this threshold. With levels <0.5 ng/ml, a correct prediction of very poor response (≤2 oocytes) was possible in 88% of cases. Levels of AMH ≥0.5 ng/ml were not significantly correlated with clinical pregnancy rates. CONCLUSIONS AMH is a predictor of ovarian response and suitable for screening. Levels ≤1.26 ng/ml are highly predictive of reduced ovarian reserve and should be confirmed by a second line antral follicle count. Measurement of AMH supports clinical decisions, but alone it is not a suitable predictor of IVF success. (Source: Human Reproduction)

Flexible gnrh antagonist versus flare-up gnrh agonist protocol in poor responders treated by ivf: a randomized controlled trial

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Although initial studies in poor responders using GnRH antagonists have reported encouraging results, they are limited in number, only a few of them are prospective, while the majority is characterized by limited power to detect a clinically important difference. METHODS A randomized controlled trial was performed in patients with one or more previous failed IVF cycles in which five or less oocytes were retrieved, using ≥300 IU of gonadotrophins/day. Patients were randomized by computer-generated list and treated by either the flare-up GnRH agonist protocol (n = 90) or a flexible GnRH antagonist protocol (n = 180). RESULTS Ongoing pregnancy rate, the primary outcome measure, was significantly higher in the antagonist group compared with the agonist group (12.2 versus 4.4%, P< 0.048; difference 7.8%, 95% CI: 0.2 to 14.0). Estradiol levels on the day of hCG administration were lower in the antagonist protocol [median (interquartile range): 572 (325–839) versus 727 (439–1029) pg/ml, P = 0.018]. Clinical and biochemical pregnancy rates, fertilization and implantation rates, as well as the number of oocytes retrieved, the number of mature oocytes present, the stimulation period and the gonadotrophin dosage were not significantly different between the two groups compared. CONCLUSIONS The flexible GnRH antagonist protocol is associated with significantly higher ongoing pregnancy rates compared with the flare-up GnRH agonist protocol in poor responders (www.clinicaltrials.gov; NCT00417066). (Source: Human Reproduction)

Estrogen addition to progesterone for luteal phase support in cycles stimulated with gnrh analogues and gonadotrophins for ivf: a systematic review and meta-analysis

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The purpose of the present systematic review and meta-analysis was to examine whether the probability of pregnancy is increased by adding estrogen to progesterone for luteal phase support in patients treated by in vitro fertilization (IVF). METHODS A literature search covering MEDLINE, EMBASE, CENTRAL, meeting proceedings and reference lists of published articles was performed to identify relevant RCTs. Data were extracted for meta-analysis yielding pooled relative risks (RR) and 95% confidence intervals (CI). Sensitivity analyses by including studies with pseudo-randomization or unclear method of randomization were also performed (n=1141 patients in total). RESULTS Four RCTs (n=587 patients) were eligible for inclusion. No statistically significant differences were present between patients who received a combination of progesterone and estrogen for luteal support when compared with those who received only progesterone, in terms of positive hCG rate (RR: 1.02, 95% CI: 0.87–1.19), clinical pregnancy rate (RR: 0.94, 95% CI: 0.78–1.13) and live birth rate (RR: 0.96, 95% CI: 0.77–1.21) per woman randomized. These results did not materially differ in the sensitivity analyses performed. CONCLUSIONS The currently available evidence suggests that the addition of estrogen to progesterone for luteal phase support does not increase the probability of pregnancy in IVF. However, there is an obvious need for further RCTs that will assess, with more confidence, the effect of estrogen addition to progesterone during the luteal phase on the probability of pregnancy. (Source: Human Reproduction)

The cost-effectiveness of long-acting reversible contraceptive methods in the uk: analysis based on a decision-analytic model developed for a national institute for health and clinical excellence (nice) clinical practice guideline

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Long-acting reversible contraceptive (LARC) methods are highly effective in preventing unintended pregnancies. However, their uptake is low in much of the developed world. This study aimed at assessing the cost-effectiveness of LARC methods from the British National Health Service (NHS) perspective. METHODS A decision-analytic model was constructed to estimate the relative cost-effectiveness of the copper intrauterine device (IUD), the levonorgestrel intrauterine system (LNG-IUS), the etonogestrel subdermal implant and the depot medroxyprogesterone acetate injection (DMPA). Comparisons with the combined oral contraceptive pill (COC) and female sterilization were also performed. Effectiveness data were derived from a systematic literature review. Costs were based on UK national sources and expert opinion. RESULTS LARC methods dominated COC (i.e. they were more effective and less costly). Female sterilization dominated LARC methods beyond 5 years of contraceptive protection. DMPA and LNG-IUS were the least cost-effective LARC methods. The incremental cost-effectiveness ratio of implant (most effective LARC method) versus IUD (cheapest LARC method) was £13 206 per unintended pregnancy averted for 1 year of use and decreased until implant dominated IUD in 15 years. Discontinuation was a key determinant of the cost-effectiveness of LARC methods. CONCLUSIONS LARC methods are cost-effective from the British NHS perspective. Practices improving user satisfaction and continuation of LARC method use should be identified and promoted. (Source: Human Reproduction)

Isolation of human single chain variable fragment antibodies against specific sperm antigens for immunocontraceptive development

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Contraceptive vaccines can provide valuable alternatives to current methods of contraception. We describe here the development of sperm-reactive human single chain variable fragment (scFv) antibodies of defined sperm specificity for immunocontraception. METHODS Peripheral blood leukocytes (PBL) from antisperm antibody-positive immunoinfertile and vasectomized men were activated with human sperm antigens in vitro, and the complementary DNA prepared and PCR-amplified using primers based on all the variable regions of heavy and light chains of immunoglobulins. The scFv repertoire was cloned into pCANTAB5E vector to create a human scFv antibody library. RESULTS Panning of the library against specific sperm antigens yielded several clones, and the four strongest reactive were selected for further analysis. These clones had novel sequences with unique complementarity-determining regions. ScFv antibodies were expressed, purified and analyzed for human sperm reactivity and effect on human sperm function. AFA-1 and FAB-7 scFv antibodies both reacted with fertilization antigen-1 antigen, but against different epitopes. YLP20 antibody reacted with the expected human sperm protein of 48 ± 5 kDa. The fourth antibody, AS16, reacted with an 18 kDa sperm protein and seems to be a human homologue of the mouse monoclonal recombinant antisperm antibody that causes sperm agglutination. All these antibodies inhibited human sperm function. CONCLUSIONS This is the first study to report the use of phage display technology to obtain antisperm scFv antibodies of defined antigen specificity. These antibodies will find clinical applications in the development of novel immunocontraceptives, and specific diagnostics for immunoinfertility. (Source: Human Reproduction)

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Induced abortions previous to ivf: an epidemiologic register-based study from finland

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The purpose of this study was to identify how many women treated for infertility had an abortion history, as well as when those abortions were carried out, and for what reasons. METHODS Data on all women treated in Finland from 1996–1998 for infertility either with IVF (n = 9175) or ovulation induction (OI, n = 10 254) and the age-matched controls of IVF women were linked to the Abortion and Hospital Discharge Registers for the period 1969–2000. RESULTS A notable proportion of IVF women (12%) and OI women (11%) had previous induced abortion(s). Practically all abortions were for social or age reasons. Most IVF women (72% of n = 1099) had their most recent abortion more than 10 years previous to fertility treatment, but more recently among OI women (45% of n = 1123 of the most recent abortions were in the preceding 10 years). Many IVF- and OI women were young and single at the time of the most recent abortion. At the time of IVF treatment most women were aged over 30 and married; OI women were also frequently married, but 42% of them were aged younger than 30. CONCLUSIONS At different points in their life, women may rely on opposite fertility regulation strategies. Health care professionals providing IVF need to consider the possibility of a previous abortion. Young women need information on the possibility of future infertility in later age. (Source: Human Reproduction)

The impact of a decline in fecundity and of pregnancy postponement on final number of children and demand for assisted reproduction technology

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Over the past decades, the proportion of couples who resort to infertility treatment has tremendously increased, and fertility (the final number of children) sharply declined. We explored the roles of two potential causes of these trends: a temporal decline in the couples' fecundability and a postponement of age at initiation of pregnancy attempts. METHODS We conducted a Monte–Carlo simulation for the reproductive history of 100 000 women based on the fertility and socio-demographic characteristics of the 1968 birth cohort in France. Declines in fecundability of various amplitudes have been implemented, as well as increases in the distribution of age at initiation of pregnancy attempts. RESULTS A decline in fecundability by 15% implied a decrease in fertility by 4%, and an increase in the proportion of couples eligible for infertility treatments by 73%. An increase in the mean age at initiation of first pregnancy attempt by 2.5 years from 25 years entailed a decrease by 5% in fertility and an increase by 32% in the proportion of couples eligible for infertility treatments. CONCLUSION A relatively important decrease in fecundability and an increase by 2.5 years in age at first pregnancy attempt are likely to have only a limited impact on fertility. However, they may have a large impact on the proportion of involuntarily infertile couples, likely to resort to assisted reproduction techniques. (Source: Human Reproduction)

Congenital anomalies in twins: a register-based study

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The risk of congenital anomalies in twins is higher than in singletons, but it is less well reported in relation to chorionicity. The aim of this study was to describe the prevalence of congenital anomalies in twin pregnancies by chorionicity and by major subtype and compare the rates with those in singletons. METHODS The study population included 2329 twin pregnancies (4658 twins) and 147 655 singletons delivered in the Northeast of England during 1998–2002. Data were obtained from the population-based Northern Multiple Pregnancy Register and Northern Congenital Abnormality Survey. RESULTS The rate of congenital anomalies in twins was 405.8 per 10 000 twins versus 238.2 per 10 000 singletons [rate ratios (RR) = 1.7, 95% confidence interval (CI) 1.5–2.0]. In twins with known chorionicity (84.8% of all twins), the prevalence of congenital anomalies in monochorionic (MC) twins (633.6 per 10 000) was nearly twice that in dichorionic (343.7 per 10 000; RR = 1.8, 95% CI 1.3–2.5). There was an increased rate of congenital anomalies in twin compared with singleton pregnancies for all major types of anomalies, except chromosomal abnormalities. CONCLUSIONS This study using high quality, population-based data on multiple pregnancies and congenital anomalies found that twins, particularly MC twins, have a higher risk of congenital anomalies than singletons. (Source: Human Reproduction)

Admission to hospital of singleton children born following assisted reproductive technology (art)

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Adverse perinatal outcomes are more common in singletons born following assisted reproductive technology (ART) and this would predict an increase in hospitalization during infancy and early childhood. METHODS We investigated hospital admissions during the first 3 years of life for all singleton children born in Western Australia between 1994 and 2000 [1328 ART, 162 350 spontaneously conceived (SC)]. RESULTS ART infants had a significantly longer birth admission and were four times more likely to be admitted to neonatal intensive care units (NICU) than SC infants. ART children had a 60% greater risk of one or more admissions in their first year and an equal risk of admission in their second and third years. Their length of stay in hospital was longer in each age period. Maternal, infant and socio-economic confounders accounted for most of the increased admission risk in the first year. However, after adjustment, a 20% increase in the risk of admission to NICU (P < 0.05) and admission to hospital during the first year (P < 0.05) remained. CONCLUSIONS Couples undertaking ART should be aware that ART infants are more likely to be admitted to a NICU, to be hospitalized in the first year of life and to stay in hospital longer than other children. (Source: Human Reproduction)

Paternal age and adverse birth outcomes: teenager or 40+, who is at risk?

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Most previous studies on the effect of paternal age have focused on the association of advanced paternal age with congenital anomalies. The objective of this study was to determine whether paternal age is associated with the risk of adverse birth outcomes, independent of maternal confounders. METHODS We carried out a retrospective cohort study of 2 614 966 live singletons born to married, nulliparous women aged 20–29 years between 1995 and 2000 in the USA. Multiple logistic regressions were applied to estimate the independent effect of paternal age on adverse birth outcomes. RESULTS Compared with infants born to fathers aged 20–29 years, infants fathered by teenagers (<20 years old) had an increased risk of preterm birth [odds ratio (OR) = 1.15, 95% confidence interval (CI): 1.10, 1.20], low birth weight (OR = 1.13, 95% CI: 1.08, 1.19), small-for-gestational-age births (OR = 1.17, 95% CI: 1.13, 1.22), low Apgar score (OR = 1.13, 95% CI: 1.01, 1.27), neonatal mortality (OR = 1.22, 95% CI: 1.01, 1.49) and post-neonatal mortality (OR = 1.41, 95% CI: 1.09, 1.82). Advanced paternal age (≥40 years) was not associated with the risk of adverse birth outcomes. CONCLUSIONS Teenage fathers carry an increased risk of adverse birth outcomes that is independent of maternal confounders, whereas advanced paternal age is not an independent risk factor for adverse birth outcomes. (Source: Human Reproduction)

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Restricted expression of the human daz protein in premeiotic germ cells

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND The role of the Y chromosome-encoded Deleted in Azoospermia (DAZ) gene family in spermatogenesis remains unclear. The ability of men without the DAZ gene to produce sperm, as well as the lack of selective pressure on DAZ exon sequences during evolution, casts doubts on its functional significance. Most men have four DAZ genes encoding protein isoforms that differ significantly in size. However, published western blots showed only a single "DAZ" band, raising the possibility that not all four DAZ genes are expressed. METHODS RT–PCR, western blotting and immunostaining were used to study the expression of the four DAZ genes and the autosomal DAZL gene in human testes and in tissue culture cells. RESULTS RNA transcripts of all four DAZ genes were found in the testis, but at much lower levels than that of the DAZL transcripts. Expression in cultured somatic cells showed that DAZ transcripts encoding multiple DAZ repeats were translated inefficiently. No DAZ proteins could be unambiguously identified on western blots when the testicular samples from three patients without the DAZ genes were used as negative controls. Nonetheless, low levels of DAZ were detected in the cytoplasm of spermatogonia by immunostaining. CONCLUSIONS The expression of DAZ proteins in adult human testes is restricted to the spermatogonia and suggests a premeiotic role. (Source: Human Reproduction)

Fas receptor is not present on ejaculated human sperm

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Apoptosis appears to have an essential role in the control of testis germ cell number and Fas expression has been reported in apoptotic spermatocytes and spermatids. We investigated if Fas (CD95) was present on ejaculated human sperm and any relationship between Fas on sperm and the apoptotic marker Syto16. METHODS Semen samples from 77 male partners of infertile couples were evaluated. Each sample was analysed both before and after semen preparation by conventional microscopical procedures and by flow cytometry (FC). A multiparameter FC analysis to assess simultaneously sperm concentration, sperm viability, sperm apoptosis, CD45 positive (leukocyte) and CD95 (Fas) positive cell concentration was carried out. A further 10 samples were studied by indirect immunofluorescence to confirm results. RESULTS The mean concentration of CD95 positive cells was very low (<1%), with no significant difference between normozoospermic and non-normozoospermic men. There was no correlation between apoptotic sperm and CD95 positive cell concentration. A linear correlation was found between CD95 positive cell and leukocyte (CD45 positive) concentration (r = 0.9946, P < 0.0001). CD95 mean fluorescence intensity of leukocytes was 10-fold greater than that of sperm and of isotypic control. Both incubation with activating anti-Fas antibody and betulinic acid induced apoptosis in leukocytes. Incubation with betulinic acid, but not with activating anti-Fas antibody, induced apoptosis in sperm. Pre-incubation with neutralizing anti-Fas antibody suppressed CD95 expression on leukocytes, whereas it did not change sperm CD95 peak fluorescence. CONCLUSIONS There is no detectable quantity of Fas on human ejaculated sperm. (Source: Human Reproduction)

Nuclear organization in human sperm: preliminary evidence for altered sex chromosome centromere position in infertile males

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

BACKGROUND Many genetic defects with a chromosomal basis affect male reproduction via a range of different mechanisms. Chromosome position is a well-known marker of nuclear organization, and alterations in standard patterns can lead to disease phenotypes such as cancer, laminopathies and epilepsy. It has been demonstrated that normal mammalian sperm adopt a pattern with the centromeres aligning towards the nuclear centre. The purpose of this study was to test the hypothesis that altered chromosome position in the sperm head is associated with male infertility. METHODS The average nuclear positions of fluorescence in-situ hybridization signals for three centromeric probes (for chromosomes X, Y and 18) were compared in normoozoospermic men and in men with compromised semen parameters. RESULTS In controls, the centromeres of chromosomes X, Y and 18 all occupied a central nuclear location. In infertile men the sex chromosomes appeared more likely to be distributed in a pattern not distinguishable from a random model. CONCLUSIONS Our findings cast doubt on the reliability of centromeric probes for aneuploidy screening. The analysis of chromosome position in sperm heads should be further investigated for the screening of infertile men. (Source: Human Reproduction)

Revised guidelines for good practice in ivf laboratories

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

The ‘ESHRE Guidelines for Good Practice in IVF Laboratories’ were drawn up by the Special Interest Group (SIG) in Embryology and published in the year 2000, and since then they constitute the minimal requirements for any laboratory offering assisted reproduction techniques (ART). In the understanding that the embryologist has a responsibility for the correct and justified application of ART in the laboratory, the implementation of these guidelines requires a quality management programme to be in place that encompasses and integrates the operative units, the processes and procedures that represent the core of ART clinics. In March 2004, the European Parliament issued the Directive 2004/23/EC ‘On setting standards of quality and safety for the donation, procurement, testing, processing, preservation, storage and distribution of human tissues and cells’. The Directive applies to human tissues and cells, including fresh or frozen reproductive cells for application to the human body, and is mainly concerned with increasing quality and safety through the implementation of a quality management system. Therefore, the European Society of Human Reproduction and Embryology (ESHRE) undertook a series of initiatives aiming to promote assurance of good laboratory practice and to define the concept of qualified embryologists. One ESHRE initiative was to revise the guidelines for good practice in IVF laboratories, not only in response to the need of embryologists for support and guidance in their duties, but also as a complement to the requirements issued by the Tissue and Cell Directive. The SIG in Embryology hopes that this document may assist the laboratory staff to operate according to the requirements of harmonization, implementation, inspection and certification that are now common to all European member states. (Source: Human Reproduction)

Cell identity in the preimplantation mammalian embryo: an epigenetic perspective from the mouse

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

The early preimplantation mouse embryo is a unique system where it is possible to explore the foundations of totipotency and differentiation. Following fertilization, a single cell, the zygote, will give rise to all tissues of the organism. The first signs of differentiation in the embryo are evident at the blastocyst stage with the formation of the trophectoderm, a differentiated tissue that envelopes the inner cell mass. The question of when and how the cells start to be different from each other in the embryo is central to developmental biology: as cell fate decisions are undertaken, loss of totipotency comes about. Although the blastomeres of the preimplantation embryo are totipotent, as the embryo develops some differences appear to develop between them which are, at least partially, related to the epigenetic information of each of these cells. The hypothesis of epigenetic asymmetries acting as driver for lineage allocation is presented. Although there are now some indications that epigenetic mechanisms are involved in cell fate determination, much work is needed to discover how such mechanisms are set in play upon fertilization and how they are transmitted through cell division. These considerations are further discussed in the context of preimplantation genetic diagnosis: does it matter to the embryo which cell is used for genetic diagnosis? The exquisite complexity and richness of chromatin-regulated events in the early embryo will certainly be the subject of exciting research in the future. (Source: Human Reproduction)

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Coming soon to your clinic: high-quality art

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

The concept of ‘patient-friendly’ medically assisted reproduction includes a robust set of clinical practice principles, to improve the quality of subfertility care. This concept is an important move away from the sole focus on effectiveness and high pregnancy rates in assisted reproduction technology (ART). Although the concept of ‘patient-friendly ART’ has several strong points, we feel it is incomplete. For achieving true high-quality ART, the concept should be extended to two more dimensions: timeliness and patient centredness. Moreover, we propose a change in the concept's name to the less ambiguous ‘high-quality ART’. (Source: Human Reproduction)

The human sperm proteome: the potential for new biomarkers of male fertility and a transformation in our understanding of the spermatozoon as a machine: commentary on the article 'identification of proteomic differences in asthenozoospermic sperm samples' by martinez et al.

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

(Source: Human Reproduction)

Editor's choice

Human Reproduction — Fri, 16 May 2008 04:00:00 +0100

(Source: Human Reproduction)

Parental origin of chromatin in human monopronuclear zygotes revealed by asymmetric histone methylation patterns, differs between ivf and icsi

Molecular Reproduction and Development — Fri, 16 May 2008 04:00:00 +0100

In mouse zygotes, many post-translational histone modifications are asymmetrically present in male and female pronuclei. We investigated whether this principle could be used to determine the genetic composition of monopronuclear human zygotes in conventional IVF and ICSI. First we determined whether male female asymmetry is conserved from mouse to human by staining polypronuclear zygotes with antibodies against a subset of histone N-tail post-translational modifications. To analyze human monopronuclear zygotes, a modification, H3K9me3, was selected that is present in the maternal chromatin. After IVF a total of 45 monopronuclear zygotes were obtained. In 39 (87%) of zygotes a nonuniform staining pattern was observed, proof of a bi-parental origin and assumed to result into a diploid conception. Two zygotes showed no staining for the modification, indicating that the single pronucleus was of paternal origin. Four zygotes contained only maternally derived chromatin. ICSI-derived monopronuclear zygotes (n = 33) could also be divided into three groups based on the staining pattern of their chromatin: (1) of maternal origin (n = 15), (2) of paternal origin (n = 8) or (3) consisting of two chromatin domains as dominating in IVF (n = 10). Our data show that monopronuclear zygotes originating from IVF generally arise through fusion of parental chromatin after sperm penetration. Monopronuclear zygotes derived from ICSI in most cases contain uni-parental chromatin. The fact that chromatin was of paternal origin in 24% of ICSI and in 4% of the IVF zygotes confirms earlier results obtained by FISH on cleavage stages. Our findings are of clinical importance in IVF and ICSI practice. Mol. Reprod. Dev. © 2008 Wiley-Liss, Inc. (Source: Molecular Reproduction and Development)

Selective er{alpha} activation disrupts sex organ differentiation and induces expression of vitellogenin ii and very low-density apolipoprotein ii in japanese quail embryos.

Reproduction — Thu, 15 May 2008 04:00:00 +0100

Selective ER{alpha} activation disrupts sex organ differentiation and induces expression of vitellogenin II and very low-density apolipoprotein II in Japanese quail embryos.

Reproduction. 2008 May 15;

Authors: Mattsson A, Olsson J, Brunström B

The Japanese quail (Coturnix japonica) is a widely-used model species for studying the roles of steroid hormones in avian sex differentiation. The aim of the present study was to elucidate the significance of estrogen receptors alpha and beta (ERalpha and ERbeta) in normal sex differentiation of the reproductive organs in the Japanese quail and in xenoestrogen-induced disruption of reproductive-organ differentiation. Real-time PCR indicated that ERalpha mRNA is expressed in both right and left gonads and Mullerian ducts in both sexes during early morphological differentiation. ERbeta-transcripts were also detected in gonads and Mullerian ducts, but at very low levels. Both receptor subtypes were expressed in the liver and may therefore mediate the expression of estrogen-regulated egg-yolk proteins. Aromatase mRNA was expressed at much higher levels in female than male gonads as early as embryonic day 5, indicating early sex differences in estrogen synthesis. Treatment with the ERalpha-selective agonist, propyl pyrazole triol (PPT), showed that frequently-reported xenoestrogen effects such as ovotestis formation, abnormal Mullerian duct development, and hepatic expression of egg-yolk proteins were induced by selective activation of ERalpha. Taken together, our results suggest that activation of ERalpha is crucial for estrogen-dependent sex differentiation of the reproductive organs and that ERalpha mediates xenoestrogen-induced toxicity during reproductive development in birds.

PMID: 18483074 [PubMed - as supplied by publisher]

(Source: Reproduction)

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Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose.

Reproduction — Thu, 15 May 2008 04:00:00 +0100

Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose.

Reproduction. 2008 May 15;

Authors: Isachenko E, Isachenko V, Weiss J, Kreienberg R, Katkov I, Schulz M, Lulat A, Risopatrón J, Sanchez R

This study investigates the ability of sucrose to protect spermatozoa against mitochondrial damage, artificial cryo-induction of capacitation and acrosomal reaction. Spermatozoa were isolated using the swim up procedure performed using three different media: a) Human Tubal Fluid medium (HTF, control), b) HTF with 1% human serum albumin (HSA) and c) HTF with 1% HSA and 0.25 M sucrose. 30 mcl suspensions of cells from each group were dropped directly into liquid nitrogen and stored for at least 24 hrs. Cells were thawed by quickly submerging the spheres in HTF with 1% HSA at 37 degrees C with gentle agitation. Sperm motility, viability, mitochondrial membrane potential integrity, spontaneous capacitation, and acrosomal reaction were investigated. Sperm viability, acrosome reaction and capacitation were detected using double fluorescence CTC-Hoechst 33258 staining technique. Mitochondrial function was evaluated using a unique fluorescent cationic dye JC-1. The number of progressive motile spermatozoa was significantly higher in the sucrose-supplemented medium group (57.1 +/- 3.2 %, P < 0.05) compared to controls (19.4 +/- 1.9 %). The combination of HSA and sucrose (65.2 +/- 7.9 %) has a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P<0.05) compare to HSA alone (28.6 +/- 4.7 %). In conclusion, vitrification of human spermatozoa with non-permeable cryoprotectants such as human serum albumin and sucrose can effectively cryopreserve the cells without significant loss of important physiological parameters.

PMID: 18483075 [PubMed - as supplied by publisher]

(Source: Reproduction)

Nucleolar re-activation is delayed in mouse embryos cloned from two different cell lines

Molecular Reproduction and Development — Sat, 10 May 2008 04:00:00 +0100

Aim of this study was to evaluate and compare embryonic genome activation (EGA) in mouse embryos of different origin using nucleolus as a marker. Early and late 2-cell and late 4-cell stage embryos, prepared by in vitro fertilization (IVF), parthenogenetic activation (PG), and nuclear transfer of mouse embryonic fibroblast (MEF) and mouse HM1 embryonic stem cells (HM1), were processed for autoradiography following 3H-uridine incubation (transcriptional activity), transmission electron microscopy (ultrastructure) and immunofluorescence (nucleolar proteins; upstream binding factor, UBF and nucleophosmin, B23). All early 2-cell embryos showed transcriptional activity only in nucleoplasm, not over nucleolar precursor bodies (NPBs). UBF was diffusely localized to cytoplasm and B23 to cytoplasm and nucleoplasm. Late 2-cell IVF and PG embryos displayed transcription over nucleoplasm and NPBs. Ultrastructurally, the latter were developing into functional nucleoli. NT-MEF and NT-HM1 embryos displayed transcription over nucleoplasm, but not over NPBs. Development of NPBs into nucleoli was lacking. UBF was in both groups localized to nucleoplasm or distinctly to presumptive NPBs. B23 was distinctly localized to NPBs. All 4-cell embryos presented nucleoplasmic transcription and developing fibrillo-granular nucleoli. UBF and B23 were distinctly localized to nucleoli. However, whereas fully transformed reticulated fibrillo-granular nucleoli were found in IVF and PG embryos, NT-MEF and -HM1 embryos displayed early NPBs transformation. In conclusion, despite normal onset of EGA in cloned embryos, activation of functional nucleoli was one cell cycle delayed in NT embryos. NT-MEF embryos displayed normal targeting but delayed activation of nucleolar proteins. Contrary, in NT-HM1 embryos, both of these processes were delayed. Mol. Reprod. Dev. © 2008 Wiley-Liss, Inc. (Source: Molecular Reproduction and Development)

Functional characterization and expression analysis of the androgen receptor in zebrafish (danio rerio) testis.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

Functional characterization and expression analysis of the androgen receptor in zebrafish (Danio rerio) testis.

Reproduction. 2008 May 9;

Authors: de Waal P, Wang D, Nijenhuis W, Schulz R, Bogerd J

The biological activity of androgens, important for male sexual differentiation and development, is mediated by the androgen receptor that binds to specific DNA recognition sites regulating the transcription of androgen target genes. We investigated androgen production by adult zebrafish testis tissue, and identified 11beta-hydroxyandrostenedione, 11-ketoandrostenedione and 11-ketotestosterone as main products, and hence potential ligands, for the zebrafish androgen receptor (Ar). These androgens were then included in the pharmacological characterization of the zebrafish Ar. The zebrafish Ar responded well in terms of binding and transactivation to synthetic androgens as well as to testosterone and 11-ketotestosterone, and reasonably well to 11-ketoandrostenedione and androstenedione. In situ hybridization analysis of zebrafish testis revealed that ar mRNA expression was detected in the subpopulation of Sertoli cells contacting early spermatogonia.

PMID: 18469035 [PubMed - as supplied by publisher]

(Source: Reproduction)

Spatio-temporal expression pattern of progranulin in embryo implantation and placenta formation suggests a role in cell proliferation, remodelling and angiogenesis.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

Spatio-temporal expression pattern of progranulin in embryo implantation and placenta formation suggests a role in cell proliferation, remodelling and angiogenesis.

Reproduction. 2008 May 9;

Authors: Desmarais J, Cao M, Bateman A, Murphy B

Embryo implantation in the mink is preceded by a variable but obligate period of delay in development. Under the influence of progesterone and unknown luteal factors, the mink embryo implants 11-13 days following its exit from diapause. Recent work suggests that progranulin, a growth factor and secreted glycoprotein, is involved in trophoblast proliferation, placental development and endometrial differentiation in the mouse. Using the mink model of delayed implantation and endotheliochorial placentation, we examined the spatio-temporal distribution of progranulin in trophoblast and endometrium during pre- and early post-implantation gestation in vivo. A partial sequence of the mink progranulin gene was cloned and sequenced. Comparative sequence analysis revealed that exons 1 and 2 of mink progranulin share 86.6 %, 82.4%, 94.9% nucleic acid sequence identity with the human, mouse and dog sequences respectively, and indicated that the invariable residues of the cysteine-rich motifs of progranulin are well conserved in the mink sequence. By in situ hybridization, we show that mink progranulin transcript is present in the cytotrophoblast and in epithelial and stromal endometrial cells at the site of implantation and during early placental formation. Immunohistochemistry revealed the progranulin protein to be strongly expressed in endometrial luminal and glandular epithelium around the time of implantation. In the incipient labyrinth, progranulin expression is localised to cytotrophoblasts, fetal capillaries, as well as to the hypertrophied maternal endothelial cells. This study demonstrates that high levels of progranulin expression correspond to active cell proliferation, remodelling and angiogenesis occurring during the establishment of the placenta in the mink.

PMID: 18469036 [PubMed - as supplied by publisher]

(Source: Reproduction)

Short term maternal psychological stress in the post-conception period in ewes affects fetal growth and gestation length.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

Short term maternal psychological stress in the post-conception period in ewes affects fetal growth and gestation length.

Reproduction. 2008 May 9;

Authors: Hill J, Smith J, Ferguson D, Jauregui G, Panarace M, Medina M, Lehnert S

Fetal development can be influenced by maternal environment in the peri-conceptional period. This study investigated the effect of maternal feed intake and psychological stress within the first 6 days after conception, on embryo development and fetal growth. Superovulated ewes (n=40) were artificially inseminated with semen from one ram. Ewes were then divided into four groups (n=10). Group 1, (Control), was fed at maintenance level, group 2 (High) at 2 x maintenance and group 3 (Low) at 0.5 x maintenance on Days 2-6 after conception. Group 4 (Stress) was fed at maintenance level and then an intense physical and psychological stress challenge was applied for 1 hour only on Days 2 and 3 after conception. Embryos were recovered at Day 6. A total of 113 transferable grade embryos were transferred singly into synchronized untreated recipients while the remaining embryos (n=165) were fixed and stained for cell counts. Post conception maternal stress or feed intake did not alter the cell count or grade of Day 6 embryos. Fetuses from the stress group had longer crown rump lengths at Day 30 and longer femur length at Day 58. Fetuses from the stressed and high feed groups had greater abdominal circumferences at Day 85. Subsequent birth weights were not significantly different. Ewes carrying lambs from the stress treatment had shorter gestation lengths. These results show that short term perturbations of the post-conception maternal environment, have measurable effects on fetal development and gestation length.

PMID: 18469037 [PubMed - as supplied by publisher]

(Source: Reproduction)

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He6/gpr64 adhesion receptor co-localizes with apical and subapical f-actin scaffold in male excurrent duct epithelia.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

HE6/GPR64 adhesion receptor co-localizes with apical and subapical F-actin scaffold in male excurrent duct epithelia.

Reproduction. 2008 May 9;

Authors: Kirchhoff C, Samalecos A, Osterhoff C

A role for HE6/Gpr64 in male excurrent ducts in the regulation of water balance was suggested from targeted gene mutation in the mouse. Results of the present immunolocalization study strengthen this hypothesis. Employing monospecific antibodies and laser confocal microscopy we studied the localization of the receptor protein in the human and wild-type mouse ductuli efferentes and epididymis. We show that HE6/Gpr64 is specifically associated with cell types and subcellular domains involved in the process of fluid re-absorption. In the mouse, dual labelling with anti-tubulin antibodies revealed that HE6/Gpr64 was absent from the (kino-)cilia of ciliated cells and also from narrow or apical cells. Instead, the receptor protein accumulated in the non-ciliated principal cells. Specifically strong immunofluorescence was observed in the apical compartment of these cells. Dual labelling with phalloidin and anti-ezrin antibodies revealed that in the mouse the bulk amount of HE6/Gpr64 protein co-localized with the F-actin-ezrin scaffold in brush border-like microvilli of ductuli efferentes and long stereocilia of the epididymis proper. In the ductuli efferentes, HE6/Gpr64 also co-localized with the subapical F-actin network immediately below the microvilli. Comparable immunostaining patterns were observed in human and mouse; however, a specific feature of the human ductuli efferentes was an intense HE6/Gpr64-related labelling of crypt-like grooves or furrows of hitherto unknown function.

PMID: 18469038 [PubMed - as supplied by publisher]

(Source: Reproduction)

Follicular development in european ground squirrels (spermophilus citellus) in different phases of the annual cycle.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

Follicular Development in European Ground Squirrels (Spermophilus citellus) in Different Phases of the Annual Cycle.

Reproduction. 2008 May 9;

Authors: Millesi E, Strauss A, Burger T, Hoffmann I, Walzl M

In seasonally breeding mammals, in particular hibernators, reproduction underlies severe energetic and temporal constraints to enable the allocation of sufficient body-fat reserves before winter. Thus, the timing of conception in spring can be crucial in terms of reproductive success. This study investigates follicular development in European ground squirrels (Spermophilus citellus) in three phases of the annual cycle: at vernal emergence, after weaning the offspring and shortly before hibernation. The animals were kept in outdoor enclosures within the natural habitat of the species. They were captured in weekly intervals, weighed and reproductive status was determined. Unilateral ovariectomy was scheduled such that the three periods were sampled. Numbers and diameters of tertiary follicles and corpora lutea in each ovary were determined, and plasma oestradiol and progesterone levels were analysed. The highest numbers of tertiary follicles, including Graafian follicles, were found in ovaries at spring emergence. During postlactation the number of tertiary follicles was lower, and active corpora lutea appeared in the investigated ovaries. Shortly before hibernation, active corpora lutea were present, but luteolysis had started in some individuals. Both oestradiol and progesterone secretion peaked after the termination of lactation and decreased before hibernation. The results demonstrate a second oestrus cycle in European ground squirrels after weaning, including an active luteal phase. This non-reproductive oestrus cycle with its endocrine output is an intriguing phenomenon. It may positively affect both prehibernatory fattening and reproduction in the subsequent season.

PMID: 18469039 [PubMed - as supplied by publisher]

(Source: Reproduction)

Expression of growth differentiation factor 9 (gdf-9), bone morphogenetic protein 15 (bmp-15) and anti-mullerian hormone (amh) in cultured mouse primary follicles.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

Expression of growth differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15) and anti-mullerian hormone (AMH) in cultured mouse primary follicles.

Reproduction. 2008 May 9;

Authors: Sadeu JC, Adriaenssens T, Smitz J

Growth and differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15) and anti-mullerian hormone (AMH) play an important role in the primary-to-secondary follicle transition and follicle activation in vivo. In organ culture of neonatal mouse ovaries, it was observed that significantly less primary follicles develop to the secondary stage. The objectives of this study were: 1) to compare ovarian follicular populations between organ-cultured neonatal mouse ovaries and freshly isolated age-matched control ovaries; 2) to quantify RNA levels of GDF-9, BMP-15 and AMH, in cultured primary follicles; and 3) to immunolocalize GDF-9 and AMH in cultured ovaries. Ovaries from 3-day-old (PND 3) mice were cultured for 7 or 10 days in absence or presence of FSH. Follicular populations were counted in freshly isolated 13-day-old (PND 13) ovaries and organ-cultured ovaries. Transcripts were quantified in isolated primary follicles using real-time RT-PCR, and protein expressions were localized using immunohistochemistry. The number of secondary follicles in organ cultured ovaries was significantly lower than in vivo, controls. GDF-9 and BMP-15 mRNA expression levels were similar as in controls. AMH mRNA levels were significantly (P < 0.05) lower after day 10 of culture in absence of FSH. GDF-9 and AMH proteins were respectively detected in the oocytes and granulosa cells (GC) beginning at the primary and primordial stage onward. GDF-9 and BMP-15 production in cultured primary follicles are not different from in vivo controls; hence abnormal early follicle growth was not related to a deficient transcription of these factors.

PMID: 18469040 [PubMed - as supplied by publisher]

(Source: Reproduction)

{beta}-microseminoprotein in human spermatozoa and its potential role in male fertility.

Reproduction — Fri, 09 May 2008 04:00:00 +0100

{beta}-microseminoprotein in human spermatozoa and its potential role in male fertility.

Reproduction. 2008 May 9;

Authors: Franchi N, Avendaño C, Molina R, Tissera A, Maldonado C, Oehninger S, Coronel C

Beta-microseminoprotein (MSMB) is one of the most abundant proteins in human seminal plasma. The objectives of this study were: (1) to purify MSMB from seminal plasma (SP) and generate antibodies against the pure protein; (2) to investigate the interaction of MSMB with ejaculated spermatozoa and its possible effect on the spontaneous acrosome reaction (AR) and (3) to quantify MSMB content in seminal plasma and examine its relationship with the clinical sperm parameters. MSMB was purified from SP and its presence on the sperm surface was examined by indirect immunofluorescence using a specific polyclonal antibody. The effect of MSMB on the AR was evaluated using guinea pig epididymal spermatozoa as model. MSMB quantification assay was performed with a two-site binding enzyme immunoassay (ELISA) using two polyclonal antibodies against MSMB. MSMB was assessed in semen samples from fertile donors (controls) and from subfertile patients according to WHO criteria. MSMB was detected on the sperm surface and mainly localized to the acrosomal region of the head and neck. A significant spontaneous acrosome reaction inhibition was observed when guinea pig epididymal spermatozoa were preincubated with MSMB. Finally, MSMB was significantly increased in subfertile patients when compared with fertile controls (p < 0.02). The association of MSMB to the sperm surface, the inhibitor effect on the spontaneous acrosome reaction and the increased MSMB levels found in seminal plasma in subfertile men suggests a relationship between this protein and semen quality and a possible role in the process of fertilization.

PMID: 18469041 [PubMed - as supplied by publisher]

(Source: Reproduction)

Locally elevated leukemia inhibitory factor in the inflamed fallopian tube resembles that found in tubal pregnancy.

Fertility and Sterility — Fri, 09 May 2008 04:00:00 +0100

Locally elevated leukemia inhibitory factor in the inflamed fallopian tube resembles that found in tubal pregnancy.

Fertil Steril. 2008 May 9;

Authors: Ji YF, Chen LY, Xu KH, Yao JF, Shi YF

OBJECTIVE: To investigate the association between the expressions of leukemia inhibitory factor and the occurrence of tubal pregnancy. DESIGN: Prospective observational study. SETTING: University-based obstetrics and gynecology hospital. PATIENT(S): Thirty women undergoing salpingectomy for tubal pregnancy and 30 nonpregnant patients with benign uterine or appendix disease. INTERVENTION(S): Oviduct tissues with ectopic gestation were separated into implantation sites and nonimplantation sites. Samples of ampullary fallopian tubes during midsecretory phase were collected as control groups. Immunohistochemical and Western blot analysis were performed. MAIN OUTCOME MEASURE(S): The differences of leukemia inhibitory factor expression between the implantation and nonimplantation sites of oviduct tissues and the normal and chronically inflamed fallopian tubes. RESULT(S): The expression of leukemia inhibitory factor in the implantation group is significantly higher than that in the nonimplantation group or in the normal group. A statistically significant difference was also found for leukemia inhibitory factor between the chronic inflammation group and the normal group by Western blot analysis but no difference between the chronic inflammation group and the implantation group or the nonimplantation group. CONCLUSION(S): Leukemia inhibitory factor might be one of the reasons that cause patients with salpingitis to be more susceptible to tubal pregnancy and might be involved in the implantation process of tubal pregnancy.

PMID: 18468598 [PubMed - as supplied by publisher]

(Source: Fertility and Sterility)

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Mechanism of severe ovarian hyperstimulation syndrome.

Fertility and Sterility — Fri, 09 May 2008 04:00:00 +0100

Mechanism of severe ovarian hyperstimulation syndrome.

Fertil Steril. 2008 May 9;

Authors: Manno M, Tomei F

PMID: 18468599 [PubMed - as supplied by publisher]

(Source: Fertility and Sterility)

Role of human albumin in ovarian hyperstimulation syndrome.

Fertility and Sterility — Fri, 09 May 2008 04:00:00 +0100

Role of human albumin in ovarian hyperstimulation syndrome.

Fertil Steril. 2008 May 9;

Authors: Cohen BM

PMID: 18468600 [PubMed - as supplied by publisher]

(Source: Fertility and Sterility)

Sensory nerve endings and endometriotic implants.

Fertility and Sterility — Fri, 09 May 2008 04:00:00 +0100

Sensory nerve endings and endometriotic implants.

Fertil Steril. 2008 May 9;

Authors: Fraser IS, Tokushige N, Markham R, Russell P

PMID: 18468601 [PubMed - as supplied by publisher]

(Source: Fertility and Sterility)

Mechanisms of severe ohss.

Fertility and Sterility — Fri, 09 May 2008 04:00:00 +0100

Mechanisms of Severe OHSS.

Fertil Steril. 2008 May 9;

Authors: Ata B, Yakin K, Alatas C, Urman B

PMID: 18468602 [PubMed - as supplied by publisher]

(Source: Fertility and Sterility)