Silver-Russell syndrome in a girl born after <Emphasis Type="Italic">in vitro</Emphasis> fertilization: partial hypermethylation at the differentially methylated region of <Emphasis Type="Italic">PEG1</Emphasis>/<Emphasis Type="Italic">MEST</Emphasis>

Purpose: The prevalence of low birth weight (LBW) is increased in subjects born after assisted reproduction technology (ART), and defective imprinting has frequently been identified in patients with Beckwith-Wiedermann and Angelman syndromes conceived by ART. Thus, we examined methylation pattern in a girl born after ART who had Silver-Russell syndrome (SRS) which can be caused by maternal uniparental disomy for chromosome 7 and by hypomethylation of the differentially methylated region (DMR) of H19. Methods: We examined methylation status of 31 cytosines at the CpG dinucleotides in the DMR of PEG1/MEST on 7q32.2 and 23 cytosines at the CpG dinucleotides in the DMR of H19 on 11p15, using leukocyte genomic DNA. Results: Eight of the 31 cytosines in the patient and four of the 31 cytosines in the father were hypermethylated in the PEG1/MEST-DMR. In the H19-DMR, no abnormal methylation pattern was identified in the patient. Conclusion: The results suggest that hypermethylation of paternally expressed genes including PEG1/MEST, which usually have growth-promoting effects, may be relevant to LBW in subjects conceived by ART. Partial hypermethylation was identified at the differentially methylated region of paternally expressed PEG1/MEST in a girl with Silver-Russell syndrome born after in vitro fertilization.     

Prevalance of <Emphasis Type="Italic">Chlamydia trochomatis</Emphasis>, <Emphasis Type="Italic">Ureaplasma urealyticum</Emphasis> and <Emphasis Type="Italic">Mycoplasma hominis</Emphasis> infections in the unexplained infertile women

Objective To prospectively investigate the prevalence of Chlamydia trachomatis (CT), Mycoplasma hominis (MH) and Ureaplasma urealyticum (UU) in the cervical canal and pouch of Douglas in unexplained infertile women and compare it to healthy controls in the Turkish population. Materials and methods A total of 31 women presenting with a history of infertility [n = 24 (77%) primary infertility, n = 7 (23%) secondary infertility] between 20 and 38 years of age and 31 women willing to have tubal ligation between 30 and 41 years of age were consecutively included into this study. Specimens were taken from intra-abdominal washings and from the cervical canal. CT, MH and UU were detected with polymerase chain reaction (PCR). Results Results of 62 women were analyzed. None of the participants met the criteria for salpingitis during laparoscopy. The most common infection in the cervical canal in both groups was UU, which was detected in 13 cases of infertile patients and 11 controls (P = 0.602). Cervical chlamydial and mycoplasmic infection was detected in one case each in infertile and control patients. Neither MH nor UU were obtained from the pouch of Douglas in both groups. Only CT was present in peritoneal fluid of an infertile woman who had also a concomitant chlamydial infection in the cervical canal. Conclusion Demonstration of cervical colonization of CT by PCR may be a promising method for the detection of asymptomatic pelvic infection in patients with unexplained infertility. However, screening for MH and UU is not cost-effective due to similar low rates of detection.     

<Emphasis Type="Italic">FTO</Emphasis>, <Emphasis Type="Italic">GCKR</Emphasis>, <Emphasis Type="Italic">CDKAL1</Emphasis> and <Emphasis Type="Italic">CDKN2A/B</Emphasis> gene polymorphisms and the risk of gestational diabetes mellitus: a meta-analysis

Purpose

Studies had examined the associations between genetic polymorphisms and the risk of gestational diabetes mellitus (GDM). However, conclusions of these studies were controversial due to the smaller sample size and limited statistical power. We carried out a meta-analysis with the aim of providing a more comprehensive summary of the currently available research to evaluate the relationship between FTO, GCKR, CDKAL1 and CDKN2A/B gene polymorphisms and GDM risk.

Methods

Literature search was carried out in the PubMed, EMBASE, Web of Science, China National Knowledge Infrastructure and Wangfang databases up to November 2017. Data were extracted by two independent reviewers and statistical analyses were performed with STATA software. Pooled odds ratios and 95% confidence intervals were calculated by Z test to assess the association between genetic polymorphisms and GDM risk. Stratified analysis was performed based on ethnicity. Heterogeneity and publication bias between studies were evaluated by Cochran’s Q test and Egger regression test, respectively.

Results

14 eligible studies were included. CDKAL1 rs7754840 and rs7756992 showed significant correlation with GDM risk under the allele, recessive, dominant, homozygote and heterozygote models. GCKR rs780094 and CDKN2A/B rs10811661 also showed the same association under the allele, recessive and heterozygote models. No associations between FTO rs9939609 and rs8050136, GCKR rs1260326 and GDM risk were found.

Conclusions

Our meta-analysis showed that two SNPs in particular(rs7754840 and rs7756992 in CDKAL1) were very strongly associated with GDM risk. GCKR rs780094 and CDKN2A/B rs10811661 polymorphisms were moderately associated with GDM risk.

     

Risk-reducing salpingo-oophorectomy in <Emphasis Type="Italic">BRCA1</Emphasis> and <Emphasis Type="Italic">BRCA2</Emphasis> mutation carriers

Background  

Risk-reducing salpingo-oophorectomy (RRSO) is often recommended to carriers of deleterious breast cancer gene 1/2 (BRCA1/2) mutations in order to reduce their breast cancer risk by 50% and their ovarian cancer risk by approximately 95%. To evaluate the acceptance, timing, histopathology findings and follow-up results we retrospectively analyzed a cohort of BRCA1/2 mutation carriers who underwent risk-reducing salpingo-oophorectomies.     

Changes in the amount of cytoplasmic inclusions in mouse oocytes during meiotic maturation <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

Background and aims: The changes in cytoplasmic inclusions during meiotic maturation have only been examined in porcine oocytes. In the present study, the amount and the number of cytoplasmic inclusions (glycogen granules, lipid droplets and fibrous structures) were examined in mouse oocytes in the process of in vivo and in vitro maturation. For those inclusions that changed in amount during maturation, we also examined their content in oocytes treated with olomoucine, an inhibitor of cyclin-dependent kinase, in order to clarify the relationship between nuclear maturation and changes in the inclusions.
Methods: Nuclear maturation in the oocytes cultured for various periods and those collected from antral follicles and oviducts was examined after staining with aceto-orcein. For the demonstration of glycogen granules and lipid droplets, oocytes were stained with periodic acid-Schiff or Sudan IV. Fibrous structures in the oocytes were observed under an electron microscope.
Results: The amount of glycogen granules, Sudanophilic lipid droplets and fibrous structures did not change in the oocytes matured in vivo and in vitro , whereas the number of the lipid droplets increased during maturation. In the oocytes treated with olomoucine, the resumption of nuclear maturation was inhibited, whereas the increase in the number of Sudanophilic lipid droplets was not inhibited.
Conclusion: Present findings suggest that the increase in the number of Sudanophilic lipid droplets occurs in the cytoplasm of mouse oocytes during maturation, regardless of in vivo or in vitro maturation, and that such the change in the inclusion is not related to nuclear maturation. (Reprod Med Biol 2004; 3 : 231–236)     

<Emphasis Type="Italic">In vitro</Emphasis> fertilization outcome in severe ovarian hyperstimulation syndrome: An age-matched contemporaneous control study

Aim:   Ovarian hyperstimulation syndrome (OHSS) is a potentially life-threatening, iatrogenic complication of assisted reproduction and has been associated with poor in vitro fertilization outcome. The aim of the present study was to evaluate the pregnancy rate and outcome following severe OHSS, at a single center over a three-year period.
Methods:   The incidence of severe OHSS at the IVF Center, National University of Singapore, in Singapore, was 4% (48 cases over 1200 cycles) during the period of 1997–2000. The present retrospective study compared 48 cases of severe OHSS to 144 age-matched, contemporaneous controls without OHSS.
Results:   The total gonadotropin required for severe OHSS group was found to be lower than for that of controls (2664.06 ± 768.29 IU vs 3349.58 ± 2003.73 IU), although duration of stimulation was similar. The OHSS group was associated with a fivefold increase (OR 5.293, 95% CI: 2.116–13.238) in pregnancy rate compared to controls (87.5% vs 56.9%; P  < 0.05). Late OHSS was more common (38/48 cases) and had a pregnancy rate of 97% per embryo transfer. There was no significant difference in the multiple pregnancy (54% vs 48%; P  > 0.05) and miscarriage rates (14% vs 7.3%; P  > 0.05) between the groups.
Conclusion:   Severe OHSS at our center were mostly late onset. The pregnancy rate was significantly higher, but multiple pregnancy and miscarriage rates were not significantly increased when compared to the age-matched contemporaneous controls. (Reprod Med Biol 2005; 4 : 207–211)     

Association of <Emphasis Type="Italic">TUSC1</Emphasis> and <Emphasis Type="Italic">DPF3</Emphasis> gene polymorphisms with male infertility

Purpose

Recently, genome-wide association studies of a Hutterite population in the USA revealed that five single nucleotide polymorphisms (SNPs) with a significant association with sperm quality and/or function in ethnically diverse men from Chicago were significantly correlated with family size. Of these, three SNPs (rs7867029, rs7174015, and rs12870438) were found to be significantly associated with the risk of azoospermia and/or oligozoospermia in a Japanese population. In this study, we investigated whether the rs10966811 (located in an intergenic region between the TUSC1 and IZUMO3 genes) and rs10129954 (located in the DPF3 gene) SNPs, previously related to family size, are associated with male infertility. In addition, we performed association analysis between rs12348 in TUSC1 and rs2772579 in IZUMO3 and male infertility.

Methods

We genotyped 145 patients with infertility (including 83 patients with azoospermia and 62 with oligozoospermia) and 713 fertile controls by PCR-RFLP technique for polymorphism. Because rs10966811 has no restriction sites, the SNP rs12376894 with strong linkage disequilibrium was selected as an alternative to rs10966811.

Results

There was a statistically significant association between rs12376894 proxy SNP of rs10966811 and oligozoospermia. Also, a statistically significant association between rs10129954 and azoospermia, and oligozoospermia was observed. When we assessed the relationship between rs12348 in TUSC1 and rs2772579 in IZUMO3 and male infertility traits, we found that rs12348 in TUSC1 was significantly associated with azoospermia and oligozoospermia, but rs2772579 in IZUMO3 was not associated with male infertility.

Conclusion

We found that the polymorphisms in TUSC1 and DPF3 displayed strong associations with male infertility.

     

The poly(A)-binding protein genes, <Emphasis Type="Italic">EPAB</Emphasis>, <Emphasis Type="Italic">PABPC1</Emphasis>, and <Emphasis Type="Italic">PABPC3</Emphasis> are differentially expressed in infertile men with non-obstructive azoospermia

Purpose

Azoospermia is one of the major causes of male infertility and is basically classified into obstructive (OA) and non-obstructive azoospermia (NOA). The molecular background of NOA still largely remains elusive. It has been shown that the poly(A)-binding proteins (PABPs) essentially play critical roles in stabilization and translational control of the mRNAs during spermatogenesis.

Methods

In the present study, we aim to evaluate expression levels of the PABP genes, EPAB, PABPC1, and PABPC3, in the testicular biopsy samples and in the isolated spermatocyte (SC) and round spermatid (RS) fractions obtained from men with various types of NOA including hypospermatogenesis (hyposperm), RS arrest, SC arrest, and Sertoli cell-only syndrome (SCO).

Results

In the testicular biopsy samples, both PABPC1 and PABPC3 mRNA expressions were gradually decreased from hyposperm to SCO groups (P?<?0.05), whereas there was no remarkable difference for the EPAB expression among groups. The expression levels of cytoplasmically localized PABPC1 and PABPC3 proteins dramatically reduced from hyposperm to SCO groups (P?<?0.05). In the isolated SC and RS fractions, the EPAB, PABPC1, and PABPC3 mRNA expressions were gradually decreased from hyposperm to SC arrest groups (P?<?0.05). Similarly, both PABPC1 and PABPC3 proteins were expressed at higher levels in the SC and RS fractions from hyposperm group when compared to the SC and RS fractions from either RS arrest or SC arrest group (P?<?0.05).

Conclusion

Our findings suggest that observed significant alterations in the PABPs expression may have an implication for development of different NOA forms.

     

<Emphasis Type="Italic">ESR1</Emphasis> and <Emphasis Type="Italic">ESR2</Emphasis> gene polymorphisms are associated with human reproduction outcomes in Brazilian women

Background

Important candidate genes involved in the ovarian response to exogenous FSH are the estrogen receptor genes (ESRs), since the effects of estrogens on follicle growth, maturation and oocyte release. It is known that some markers of ovarian stimulation can help to personalize the treatment, adjusting the dose of exogenous rFSH, thus preventing excessive wear of the patient. Inspired on this information we aimed to analyze four different polymorphisms in the estrogen receptor genes ESR1: rs2234693/T-397C (PvuII) and rs9340799/A-351G (Xbal) and ESR2: rs4986938/G1082A (RsaI) and rs1256049/A?+?1730G (AluI), and their association with assisted reproduction outcomes in Brazilian women that underwent in vitro fertilization (IVF).

Methods

A cross-sectional study was performed involving 136 infertile women less than 39 years of age with normal ovarian reserve. Patients were divided according to the same COH protocol for statistical analysis. The Taqman assay was used for PvuII and XbaI of ESR1, and RsaI and AluI of ESR2 genotyping. Serum estradiol and FSH were measured by Elisa assay.

Results

The PvuII (ESR1) TT and RsaI (ESR2) GG genotypes were associated with a longer induction period and higher doses of medication (p?<?0.03). The XbaI (ESR1) AA genotype was associated with better COH results, including a larger number of follicles, mature oocytes, embryos, and good quality embryos (p?<?0.05). The AluI GG genotype showed an association with the Ovarian Hyperstimulation Syndrome (OHSS) (p?=?0.03). According to the haplotype analysis of ER1 (PvuII/XbaI), we demonstrated that the CA combination increases by 0.68 the number of good quality embryos while the TG decreases it by 0.71 (p?=?0.04).

Conclusion

ER polymorphisms have an association with the assisted reproduction outcomes in Brazilian women.

     

Polymorphisms of <Emphasis Type="Italic">ICAM-1</Emphasis> and <Emphasis Type="Italic">IL-6</Emphasis> genes related to endometriosis in a sample of Brazilian women

Purpose

This study investigated the possibility of K469E (rs5498) and G241R (rs1799969) polymorphisms, in ICAM-1 gene, and G634C (rs1800796), in IL-6 gene, being associated with the occurrence of endometriosis in a sample of Brazilian women.

Methods

We genotyped 200 women (100 in control group and 100 in endometriosis group) by PCR-RFLP technique for G634C, K469E, and G241R polymorphisms.

Results

No significant difference was observed in genotypic frequency between control and endometriosis groups for G634C and K469E polymorphisms (p?=?0.61 and p?=?0.22, respectively). In addition, no significant difference between stages I-II and III-IV of the disease was found for both SNPs (p?=?0.63 and p?=?0.24, respectively). All individuals were wild homozygotes for G241R polymorphism.

Conclusion

This study suggests that polymorphisms K469E, G241R, and G634C are not associated with increased susceptibility to endometriosis in Brazilian women.

     

Isolation of the human <Emphasis Type="Italic">ePAB</Emphasis> and <Emphasis Type="Italic">ePABP2</Emphasis> cDNAs and analysis of the expression patterns

Purpose  Identification of the unique genes playing critical roles in human embryo cleavage. Methods  Isolation of human ePAB cDNA using human ovary cDNA libraries and mouse ePAB amino acid sequences, followed by analysis of its expression pattern in various adult tissues and stages during early oocyte development excluding ePABP2. Results  Human ePAB encodes a 330-aa protein and is located on chromosome 20q12-q13.1. The amino acid sequence is 72% homologous with that of mouse ePab. Human ePAB has only three RRMs and lacks a PABP domain; the expression pattern is nonspecific in adult tissues and detected in all stages, from oocyte to blastocyst. Human ePABP2 encodes a 282-aa protein and is located on chromosome 16q24.3. The amino acid sequence is 68% homologous with mouse ePabp2. Conclusions  We identified human ePAB and ePABP2 cDNA. Human ePAB cDNA is not expressed specific to the ovary. Biological discrepancies exist between the human and the mouse. Capsule   We successfully isolated the human ePAB and ePABP2 cDNAs and analyzed the expression patterns in humans.     

Length difference between equine <Emphasis Type="Italic">ZFX</Emphasis> and <Emphasis Type="Italic">ZFY</Emphasis> genes and its application for molecular sex determination

Purpose  

We analyzed the sex chromosome-encoding ZFX-ZFY genes and tested molecular sexing using the amplification patterns of intron 9 of ZFX-ZFY in the horse.     

MicroRNA expression in preimplantation mouse embryos from <Emphasis Type="Italic">Ped</Emphasis> gene positive compared to <Emphasis Type="Italic">Ped</Emphasis> gene negative mice

PURPOSE: The mouse preimplantation embryo development (Ped) gene product, Qa-2, influences the rate of preimplantation embryonic development and overall reproductive success. Here we investigated the expression pattern of two microRNAs, miR-125a and miR-125b, known to be involved in development in lower organisms, in preimplantation embryos from the two-cell, four-cell, eight-cell, morula, and blastocyst stages of development from the congenic B6.K1 (Ped negative) and B6.K2 (Ped positive) strains of mice. METHOD: B6.K1 and B6.K2 congenic mice differ only in the absence (B6.K1) or presence (B6.K2) of the genes encoding Qa-2 protein. We analyzed the expression of miR-125a and miR-125b in B6.K1 and B6.K2 preimplantation embryos by using real-time PCR. RESULT: We found no variability in miR-125b expression at any developmental stage in both strains. However, miR-125a expression increased during development in both strains and was ten times higher in Ped negative (B6.K1) embryos than in Ped positive (B6.K2) embryos by the blastocyst stage of development. CONCLUSION: Our results show that the absence of the Ped gene profoundly affects the level of a miRNA (miR-125a) known to regulate early development. The implication is that miR-125a is likely involved in the regulation of timing of early development in mice.     

Physical and mental development of children after <Emphasis Type="Italic">in vitro</Emphasis> fertilization and embryo transfer

Objective:  To evaluate the physical and mental development of children after in vitro fertilization (IVF) and frozen embryo transfer (FET).
Methods:  Between July 1995 and November 2003, 506 patients delivered 658 babies after IVF and ET treatment at our clinic (intracytoplasmic sperm injection (ICSI), 418; conventional IVF, (C-IVF) 125; FET, 115). A survey of the physical and mental developmental of the children was conducted by mailing questionnaires to parents. Comparisons were made between three treatment procedures, and development of singleton, twin and triplet delivery.
Results:  The response rate was 73.4% (483/658) for 324 children born after ICSI, 78 born after C-IVF, and 81 born after FET. The height and weight of assisted reproductive technology (ART) children at birth were significantly lower than that of naturally conceived babies (ART children: natural delivery; 46.8 cm, 49.0 cm and 2524 g, 3040 g, respectively). However, there was no significant difference between the singletons alone and naturally conceived children irrespective of the ART method. In addition, mental development was the same between singletons and naturally conceived children. The ART group tended to delay body development such as 'holding their head up', 'sitting up', 'crawl' to moving growth in multiple births.
Conclusion:  The physical and mental development of twins or triplets was significantly more delayed than that of naturally conceived babies, but had improved to a similar extent of the singletons after the age of 6 months. (Reprod Med Biol 2004; 3 : 63–67)     

Association of a <Emphasis Type="Italic">TDRD1</Emphasis> variant with spermatogenic failure susceptibility in the Han Chinese

Purpose

Piwi-interacting RNAs (piRNAs) are a broad group of noncoding small RNAs that have important biological functions in germline cells and can maintain genome integrity via silencing of retrotransposons. In this study, we aimed to explore the associations between genetic variants of important genes involved in piRNA biogenesis and male infertility with spermatogenic impairment.

Methods

To this end, five single-nucleotide polymorphisms (SNPs) in the ASZ1, PIWIL1, TDRD1, and TDRD9 genes were genotyped by TaqMan allelic discrimination assays in 342 cases of nonobstructive azoospermia (NOA) and 493 controls.

Results

The SNP rs77559927 in TDRD1 was associated with a reduced risk of spermatogenic impairment. The genotypes TC and TC?+?CC showed odds ratios and 95 % confidence intervals of 0.73 (0.55–0.98, P?=?0.034) and 0.73 (0.56–0.97, P?=?0.030), respectively, in patients with NOA compared with those in the controls.

Conclusion

Thus, our results provided the first epidemiological evidence supporting the involvement of TDRD1 genetic polymorphisms in piRNA processing genes in determining the risk of spermatogenic impairment in a Han Chinese population.

     

No association of <Emphasis Type="Italic">TAP1</Emphasis> and <Emphasis Type="Italic">TAP2</Emphasis> genes polymorphism with risk of cervical cancer in north Indian population

Background  Transporter associated with antigen processing (TAP), a member of the ATP-binding cassette transporter super family, is composed of two integral membrane proteins, TAP-1 and TAP-2. The TAP gene product is involved in the processing of endogenous peptides that bind to MHC class I molecules. Mutations and/or polymorphism within these genes could alter the efficacy of the immune response which might be relevant for the development of autoimmune diseases and cancer. Methods  DNA was isolated from peripheral blood sample of 200 patients with cervical cancer and 200 healthy controls. TAP1 and TAP2 allele polymorphism were determined by polymerase chain reaction. Result  Significant protective OR (OR = 0.22 95% CI = 0.09–0.51, P < 0.001-OR = 0.47, 95% CI = 0.24–0.92, P = 0.02) was observed for GG and combined AG+GG genotypes of TAP2 in patients with SCC respectively. Similarly, such genotypes (GG, AG+GG) appeared same OR for patient with cervical cancer in study group (OR = 0.12, 95% CI = 0.04–0.39-P < 0.001-OR = 0.5 ,95% CI = 0.25–0.95-P = 0.03). There was decrease risk of cervical cancer in user of oral contraceptive with AG and GG genotypes of TAP2 (OR = 0.55, 95% Cl = 0.41–0.73, P = 0.002, OR = 0.09, 95% CI = 0.02–0.36, P < 0.001) respectively. In case of TAP1 gene all allelic polymorphisms showed a decrease OR in patients with cervical cancer in passive smokers and user of oral contraceptives, though, no significant Conclusion  Thus, TAP1 and TAP2 genes polymorphism are not linked to cervical carcinoma, since no association was found between a particular genotype and the disease.