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Human chorionic gonadotropin patterns in early pregnancy after assisted reproduction.

BACKGROUND: Assisted reproduction implies increased risks of pathological pregnancy, necessitating close follow up of early pregnancy. The use of serum hCG levels two and three weeks after embryo transfer for prediction of pregnancy outcome after in vitro fertilization-embryo transfer (IVF/ET) or gamete intrafallopian transfer (GIFT) was evaluated. METHODS: Three hundred and twenty-nine treatment cycles were included. Serum samples were obtained on days 14 and 21 after ET and hCG was determined by fluoroimmunoassay. Receiver operating characteristic (ROC) curves were analyzed to find cut-off levels of hCG giving maximal sensitivity and specificity, identifying a low risk group and a high risk group with regard to pathological pregnancy. RESULTS: The group of patients carrying a viable pregnancy had significantly higher hCG levels two and three weeks after ET than the group of patients carrying a pathological pregnancy. Furthermore, the daily increase in hCG was higher. In multiple gestations, the levels of hCG were significantly higher compared to singleton pregnancies. Ninety per cent of the patients with an hCG level > or = 150 IU/L 13-15 days after ET carried a pregnancy to term (the low risk group). Conversely, 50% of the patients with hCG < 150 IU/L carried a pathological pregnancy (the high risk group). CONCLUSION: A single determination of the hCG level two weeks after ET, combined with transvaginal ultrasound two to three weeks later, is a reliable follow-up. In cases of subnormal hCG levels (< 150 IU/L), a second hCG determination one week later followed by sonography is recommended.

Chorionic Gonadotropin↗

Incomplete denudation of oocytes prior to ICSI enhances embryo quality and blastocyst development.

BACKGROUND: Granulosa cells are essential mediators of oocyte maturation and fertilization. Because of the denudation of oocytes in preparation for ICSI, any potential positive effect of surplus cumulus cells (CCs) on further development would be unable to exert further effect. In order to evaluate the actual influence of adhering cumulus cells on further preimplantation development, this prospective study was carried out. METHODS: Sibling cumulus-oocyte complexes for 57 ICSI patients were split into a study group (incomplete denudation, n = 314) and a control group (complete denudation, n = 336). According to the cumulus cell pattern after partial denudation, mature gametes from the study group were further subdivided into type A oocytes, which showed several prominent CC clusters (n = 202), and type B (n = 75), which showed a more homogeneous pattern with CC covering the whole surface of the gamete. RESULTS: In immature oocytes, presence of adhered CCs led to a significant increase in resumption of meiosis (P < 0.01). Fertilization rate (P < 0.05) and ability to cleave (P < 0.01) was impaired in the study group, because of difficulties in ICSI of type B oocytes. By contrast, embryo morphology on days 2 (P < 0.01) and 3 (P < 0.05), as well as blastocyst formation, was better (P < 0.05) in the study group (55 blastocysts out of 88 zygotes) as compared to that in the control group (49/105). CONCLUSION: These data indicate that co-culture of oocytes with attached CCs may enhance preimplantation development.

Blastocyst↗

Familial clustering of impaired spermatogenesis: no evidence for a common genetic inheritance pattern.

BACKGROUND: The aetiology of impaired spermatogenesis is unknown in the majority of cases. Evidence of a contribution of genetic factors is still scarce. Therefore, the aim of our study was to assess whether male factor subfertility due to impaired spermatogenesis has a familial component and to test different genetic models of inheritance. METHODS: Cases were all men with severe idiopathic impaired spermatogenesis attending our fertility clinic from January 1998 until December 2001. Controls were all men with normozoospermia attending our fertility clinic in the same period. Family data were collected from the medical records and by additional interviews of the probands. If subfertility of a first-degree relative was mentioned, permission was sought to contact the affected family member in order to obtain all medical information available, including the results of semen analyses. RESULTS: In total, 160 patients and 285 controls were included in the analysis. Family size and number of brothers and sisters were equally distributed in both groups. In the patient group, 1.63% of the brothers who had tried to father a child were mentioned to be subfertile compared to 5.8% in the control group [odds ratio 3.18 (95% confidence interval 1.59-6.37)]. The subfertility among the brothers in the patient group was more often due to reduced semen parameters compared to the control group. The data did not fit with frequent autosomal dominant or recessive segregation. CONCLUSION: Male factor subfertility due to impaired spermatogenesis appears to cluster in families. Our data suggests that heritable genetic factors play a role in a limited number of cases. Impaired spermatogenesis is not caused by a common genetic defect, but is most likely a complex disease in which several different factors play a role.

Adult↗

Estimation of mean age at first marriage: use of a simple mathematical model.

"The present paper is an attempt to introduce a simple mathematical model to describe the age pattern of proportion never married women. The underlying model was found to give fairly close fit to an observed set of data of some 17 WFS [World Fertility Survey] countries. A mathematical formulation was then suggested in terms of the parameters in the model to estimate the mean age at first marriage. The mean ages obtained under the approach agreed quite closely with those obtained by Hajnal's method. The agreement between the estimates of ever married proportions obtained by the suggested model and...Coale's nuptiality model appeared also to be satisfactory."

Age Distribution↗

Single nucleotide polymorphisms of the heat shock protein 90 gene in varicocele-associated infertility.

PURPOSE: Varicoceles are associated with impaired testicular function and male infertility, but the molecular mechanisms by which fertility is affected have not been satisfactorily explained. Spermatogenesis might be affected by increased scrotal temperature, such as that caused by varicocele. HSP90 is a molecular chaperone expressed in germ cells and is related to spermatogenesis, motility, and both heat and oxidative stress. Possible correlations between coding single region nucleotide polymorphisms (cSNPs) in the HSP90 gene in patients with varicocele associated with infertility were analyzed, and polymorphisms in these exons were characterized through DNA sequencing. MATERIALS AND METHODS: PCR-SSCP and DNA sequencing were used to search for mutations in 18 infertile patients with varicocele, 11 patients with idiopathic infertility and 12 fertile men. DNA was extracted from leucocytes for PCR amplification and SSCP analysis. DNA from samples with an altered band pattern in the SSCP was then sequenced to search for polymorphisms. RESULTS: Three silent polymorphisms that do not lead to amino acid substitutions were identified. CONCLUSION: Mutations in the HSP90 gene do not appear to be a common cause of male factor infertility. The low incidence of gene variation, or SNPs, in infertile men demonstrates that this gene is highly conserved and thus confirms its key role in spermatogenesis and response to heat stress.

HSP90 Heat-Shock Proteins↗

Use of diamide-acridine orange fluorescence staining to detect aberrant protamination of human-ejaculated sperm nuclei.

OBJECTIVE: To investigate the influence of human sperm nuclear chromatin on fertilization. DESIGN: Prospective study. SETTING: Assisted reproductive technology unit at a university teaching hospital. PATIENT(S): Fifty men starting an IVF-ET program. INTERVENTION(S): Epifluorescent microscopic observation of human-ejaculated sperm nuclei stained with diamide-acridine orange. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of extracted sperm nucleoproteins. MAIN OUTCOME MEASURE(S): Usefulness of diamide-acridine orange in analysis of human sperm nuclear chromatin and fertilization ability. RESULT(S): There was no correlation between the semen parameters and the diamide-acridine orange observation. A positive correlation was observed between the fertilization rate after conventional IVF and the green-type increase ratio (percentage of green-pattern sperm after diamide-acridine orange staining/percentage of green-pattern sperm after acridine orange staining). Furthermore, it was suggested by SDS-PAGE that structural differences were noticed between the fertile men and the men with sperm immaturity diagnosed after diamide-acridine orange staining. CONCLUSION(S): Diamide-acridine orange staining was a more precise method for detecting chromatin abnormalities in human-ejaculated sperm and evaluating fertilization ability than acridine orange staining alone. This method can be used as a diagnostic tool to assess the fertilization ability of human-ejaculated spermatozoa before IVF procedures.

Acridine Orange↗

Cross-fertilization and structural comparison of egg extracellular matrix glycoproteins from Xenopus laevis and Xenopus tropicalis.

While the anuran amphibian Xenopus laevis is a widely used vertebrate model system, it is not optimal for genetic manipulations due to its tetraploid genome and long generation time. A current alternative amphibian model system, Xenopus tropicalis, has the advantages of a diploid genome and a much shorter generation time. We undertook a comparative investigation of X. tropicalis egg extracellular matrix glycoproteins in relation to those already characterized in X. laevis. Fertilization methods and isolation of egg extracellular molecules were directly transferable from X. laevis to X. tropicalis. Cross-fertilizations were successful in both directions, indicating similar molecules involved in sperm-egg interactions. Egg envelopes analyzed by SDS-PAGE were found to have almost identical gel patterns, whereas jelly component profiles were similar only for the larger macromolecules (>90 kDa). The cDNA sequences for egg envelope glycoproteins ZPA, ZPB, ZPC, ZPD and ZPAX, and also egg cortical granule lectin involved in the block to polyspermy, were cloned for X. tropicalis and showed a consistent approximately 85% amino acid identity to the X. laevis sequences. Thus, homologous egg extracellular matrix molecules perform the same functions, and the molecular and cellular mechanisms of fertilization in these two species are probably equivalent.

Amino Acid Sequence↗

The preovulatory serum estradiol pattern in natural IVF/ICSI cycles.

PURPOSE: The aim of the study was to find whether inferences to the possible success of natural IVF/ICSI cycles could be drawn from the estradiol (E2) pattern. METHODS: Sixty-eight women who underwent oocyte recovery in 98 natural cycles were recruited for the study. Daily serum E2 was measured in the preovulatory phase (-3 to +2 day). The E2 pattern was compared among four groups: Group A, unsuccessful egg retrieval; Group B, no fertilization; Group C, no implantation; and Group D, implantation. RESULTS: There was no difference in mean E2 levels between groups. Only the ratio of E2 on day +1/E2 on day 0 was significantly lower in conception cycles in comparison with nonconception cycles. In cycles with a decreased E2 level on day +1, only the implantation rate was significantly higher in comparison with cycles with an increasing E2 level. CONCLUSIONS: From the E2 pattern it is possible to make inferences about the likelihood of implantation but not the fertilization or oocyte recovery success.

Adult↗

Fertility and contraception in the Marshall Islands.

Data on fertility and contraception in Micronesian women in the Marshall Islands were collected during a women's health survey in 1985. High total fertility rates were found. The reproductive pattern of many Marshallese women is one that has been associated with adverse health consequences: pregnancies in teenagers and in women over 39 years, high parities of four or more births, and short birth intervals. The practice of breastfeeding is declining in younger women. The prevalence of contraceptive use is low, and the availability of reversible methods is limited. Most contraceptive nonusers would like to practice contraception, but are inhibited by the lack of information about family planning. It is suggested that more attention needs to be given to family planning services in the Marshall Islands, in particular to improving the availability of reversible methods of contraception and of information about family planning. Further research is also needed on how family planning services might best be organized to maximize participation by women and their partners who wish to use such services.

Adolescent↗

Expression of growth factor ligand and receptor genes in the preimplantation bovine embryo.

The sensitive technique of mRNA phenotyping with the reverse transcription-polymerase chain reaction was employed to determine the patterns of gene expression for several growth factor ligand and receptor genes during bovine preimplantation development. Several thousand bovine embryos encompassing a developmental series from one-cell zygotes to hatched blastocysts were produced by the application of in vitro maturation, fertilization, and oviductal epithelial cell embryo coculture methods. Transcripts for transforming growth factor (TGF-alpha) and platelet-derived growth factor (PDGF-A) are detectable in all preimplantation bovine stages as observed in the mouse. Transcripts for TGF-beta 2 and insulin-like growth factor (IGF-II) and the receptors for PDGF-alpha, insulin, IGF-I, and IGF-II are also detectable throughout bovine preimplantation development, suggesting that these mRNAs are products of both the maternal and the embryonic genomes in the cow, whereas in the mouse they are present only following the activation of the embryonic genome at the two-cell stage. In contrast to the mouse embryo, IGF-I mRNA was detected within preimplantation bovine embryos. Basic fibroblast growth factor (bFGF) is a maternal message in the bovine embryo, since it is only detectable up until the eight-cell embryo stage. Bovine trophoblast protein (bTP) mRNA was detectable within day 8 bovine blastocysts. As was observed in the mouse, the transcripts for insulin, epidermal growth factor (EGF), or nerve growth factor (NGF) were not detectable in any bovine embryo stage. Analyses of this type should aid the development of a completely defined culture medium for the more efficient production of preimplantation bovine embryos.

Animals↗

Segregation of germline granules in early embryos of Caenorhabditis elegans: an electron microscopic analysis.

Using an improved fixation method for electron microscopy, we have found germline granules in Caenorhabditis elegans embryos shortly after fertilization and prior to the first cleavage. They are localized in the egg cytoplasm which becomes segregated into the posterior blastomere at the first cleavage. In the following divisions, the granules continue this pattern of asymmetric segregation and are ultimately segregated into the germline precursor cell. The granules are then symmetrically segregated into the germline cells.

Animals↗

Genetic changes in human fetuses from spontaneous abortion after in vitro fertilization detected by comparative genomic hybridization.

The in vitro fertilization (IVF) technique is becoming a very important approach for infertile disease therapy, but approximately 30% of pregnancies are spontaneously aborted in the first trimester. It is believed that chromosomal abnormality is the major reason for early spontaneous abortion. Although some reports have mentioned cytogenetic changes in spontaneously aborted embryos after IVF, little is known about the comprehensive cytogenetic alterations in these aborted embryos. Here we use the comparative genomic hybridization (CGH) technique to analyze the genetic alterations in 41 spontaneously aborted human specimens after IVF. In this study, 25 of 41 cases (61%) showed chromosomal changes. Among them, autosomes and sex chromosomes were involved in 16 and 11 cases, respectively. Several nonrandom chromosomal changes were identified, including loss of one sex chromosome (six cases) and gains of 22 (four cases), Y (four cases), 21 (three cases), 4 (two cases), and 13 (two cases). Our data support the opinion that chromosome abnormality is one of the major causes of early spontaneous abortion after IVF. The association between chromosome changes in these spontaneously aborted fetuses and maternal age, infertility patterns, infertility causes, and IVF patterns (routine IVF and other methods, including intracytoplasmic sperm injection, egg donation, and embryo donation) were also studied. No significant correlation was found.

Abortion, Spontaneous↗

Spermatids as gametes: indications and limitations.

The feasibility of achieving viable embryos, developing to term after transfer into the uterus, by fertilizing oocytes with spermatids has been demonstrated both in animal studies and in preliminary human clinical trials. Here we review the current clinical indications of spermatid conception and discuss the predictable success rates associated with each of these indications. Potential health hazards relating to the use of spermatids for conception are updated taking into account the risk of abnormal or incomplete epigenetic modifications of newly discovered human imprinted genes. We also add new experimental data showing the occurrence of spermatids in patients lacking spermatozoa and demonstrating that round spermatids recovered from patients with complete spermiogenesis failure (no elongated spermatids or spermatozoa ever detected in the patient's history) are often deficient in the factor(s) responsible for oocyte activation. The possible consequences of this deficiency for the occurrence of abnormal fertilization patterns and for the impairment of further preimplantation and post-implantation development are discussed. It is concluded that the development of diagnostic tests to assess the intrinsic quality of spermatids, with regard to their ability to act as gametes, is urgently needed as part of pre-treatment diagnosis before infertile couples are included in a spermatid conception programme. Centres wishing to use spermatids in human assisted reproduction should also be prepared to offer adequate diagnostic methods to control genomic imprinting abnormalities in the progeny.

Female↗

Is the pattern of solid-phase gastric emptying different between genders?

BACKGROUND: In the 13C-octanoate breath test, the shape of the 13CO2 excretion curve in the ascending portion reflects a pattern of gastric emptying (GE). Recent scintigraphic studies have revealed an overall delay in solid GE in fertile women compared with men. However, it remains unknown whether women have a different GE pattern compared with men. As a symptomatic delay in solid GE is specific to the female gender, it could be hypothesized that the 13CO2 excretion curve is different in shape between genders. MATERIALS AND METHODS: Because the ascending gradient of the 13CO2 excretion curve is often biphasic, the dual function of y(t) = ( a1 . tb1 + a2 . tb2) e(-K.t) was applied to fit the breath data, where a1, b1, a2, b2, and K are constants. Assessed on the 4 h-based breath samples obtained after ingestion of a 320-kcal muffin containing 100 mg 13C-octanoate, the time versus 13CO2 excretion curve was created from 31 adult volunteers (15 men and 16 women). The curve shape was characterized by the dual function, and was compared between genders. RESULTS: In both genders, the ascending gradient exhibited the biphasic feature, characterized by an initial steep rise and the subsequent blunted increase, while the descending gradient followed the monotonous decay. The initial rise was steeper and the subsequent increase was more blunted in women than in men. CONCLUSION: Women exhibit a gender-specific pattern of the 13CO2 excretion profile. A possible explanation for this gender difference is that the post-gastric feedback regulation is more potent in women than in men.

Adult↗