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Sample records for stage embryo transfer

  1. Cleavage stage versus blastocyst stage embryo transfer in assisted reproductive technology.

    Science.gov (United States)

    Glujovsky, Demián; Farquhar, Cindy; Quinteiro Retamar, Andrea Marta; Alvarez Sedo, Cristian Roberto; Blake, Deborah

    2016-06-30

    Advances in cell culture media have led to a shift in in vitro fertilisation (IVF) practice from cleavage stage embryo transfer to blastocyst stage transfer. The rationale for blastocyst transfer is to improve both uterine and embryonic synchronicity and enable self selection of viable embryos, thus resulting in better live birth rates. To determine whether blastocyst stage (day 5 to 6) embryo transfers improve the live birth rate, and other associated outcomes, compared with cleavage stage (day 2 to 3) embryo transfers. We searched the Cochrane Gynaecology and Fertility Group Specialised Register of controlled trials, Cochrane Central Register of Controlled Trials (CENTRAL; the Cochrane Library; 2016, Issue 4), MEDLINE, EMBASE, PsycINFO, CINAHL, and Bio extracts from inception to 4th April 2016. We also searched registers of ongoing trials and the reference lists of studies retrieved. We included randomised controlled trials (RCTs) which compared the effectiveness of blastocyst versus cleavage stage transfers. We used standard methodological procedures recommended by Cochrane. Our primary outcomes were live birth and cumulative clinical pregnancy rates. Secondary outcomes were clinical pregnancy, multiple pregnancy, high order pregnancy, miscarriage, failure to transfer embryos, and embryo freezing. We assessed the overall quality of the evidence for the main comparisons using GRADE methods. We included 27 RCTs (4031 couples or women).The live birth rate following fresh transfer was higher in the blastocyst transfer group (odds ratio (OR) 1.48, 95% confidence interval (CI) 1.20 to 1.82; 13 RCTs, 1630 women, I(2) = 45%, low quality evidence) following fresh transfer. This suggests that if 29% of women achieve live birth after fresh cleavage stage transfer, between 32% and 42% would do so after fresh blastocyst stage transfer.There was no evidence of a difference between the groups in rates per couple of cumulative pregnancy following fresh and frozen

  2. Monozygotic Triplets and Dizygotic Twins following Transfer of Three Poor-Quality Cleavage Stage Embryos

    Directory of Open Access Journals (Sweden)

    Reshef Tal

    2012-01-01

    Full Text Available Background. Assisted reproductive technology has been linked to the increased incidence of monozygotic twinning. It is of clinical importance due to the increased risk of complications in multiple pregnancies in general and in monozygotic twins in particular. Case. A 29-year-old female, nulligravida underwent her first IVF cycle. Three poor-quality cleavage stage embryos were transferred resulting in monochorionic triamniotic triplets and dichorionic diamniotic twins. Selective embryo reduction was performed at 12 weeks leaving dichorionic twins. The patient underwent emergency cesarean section due to preterm labor and nonreassuring fetal heart tracing at 30 weeks of gestation. Conclusion. Our case emphasizes that even embryos with significant morphological abnormalities should be considered viable and the possibility of simultaneous spontaneous embryo splitting must be factored into determining number of embryos to transfer.

  3. Two-staged nuclear transfer can enhance the developmental ability of goat-sheep interspecies nuclear transfer embryos in vitro.

    Science.gov (United States)

    Ma, Li-Bing; Cai, Lu; Li, Jia-Jia; Chen, Xiu-Li; Ji, Feng-Yu

    2011-02-01

    The technique of interspecies somatic cell nuclear transfer, in which interspecies cloned embryos can be reconstructed by using domestic animal oocytes as nuclear recipients and endangered animal or human somatic cells as nuclear donors, can afford more opportunities in endangered animal rescue and human tissue transplantation, but the application of this technique is limited by extremely low efficiency which may be attributed to donor nucleus not fully reprogrammed by xenogenic cytoplasm. In this study, goat fetal fibroblasts (GFFs) were used as nuclear donors, in vitro-matured sheep oocytes were used as nuclear recipients, and a two-stage nuclear transfer procedure was performed to improve the developmental ability of goat-sheep interspecies clone embryos. In the first stage nuclear transfer (FSNT), GFFs were injected into the ooplasm of enucleated sheep metaphase-II oocytes, then non-activated reconstructed embryos were cultured in vitro, so that the donor nucleus could be exposed to the ooplasm for a period of time. Subsequently, in the second stage nuclear transfer, FSNT-derived non-activated reconstructed embryo was centrifuged, and the donor nucleus was then transferred into another freshly enucleated sheep oocyte. Compared with the one-stage nuclear transfer, two-stage nuclear transfer could significantly enhance the blastocyst rate of goat-sheep interspecies clone embryos, and this result indicated that longtime exposure to xenogenic ooplasm benefits the donor nucleus to be reprogrammed. The two-stage nuclear transfer procedure has two advantages, one is that the donor nucleus can be exposed to the ooplasm for a long time, the other is that the problem of oocyte aging can be solved.

  4. Influence of recipient cytoplasm cell stage on transcription in bovine nucleus transfer embryos

    DEFF Research Database (Denmark)

    Smith, S. D.; Soloy, E.; Kanka, J.

    1996-01-01

    Nucleus transfer for the production of multiple embryos derived from a donor embryo relies upon the reprogramming of the donor nucleus so that it behaves similar to a zygotic nucleus. One indication of nucleus reprogramming is the RNA synthetic activity. In normal bovine embryogenesis, the embryo...

  5. Nucleolar ultrastructure in bovine nuclear transfer embryos

    DEFF Research Database (Denmark)

    Kaňka, Jiří; Smith, Steven Dale; Soloy, Eva

    1999-01-01

    all three cell cycles. In the eight-cell stage embryo, a primary vacuole appeared as an electron lucid area originating in the centre of the nucleolar precursor body. In nuclear transfer embryos reconstructed from nonactivated cytoplasts, the nuclear envelope was fragmented or completely broken down...... vacuoles. A nucleolar precursor body typical for the two-cell stage control embryos was never observed. None of the reconstructed embryos of this group reached the eight-cell stage. Nuclear transfer embryos reconstructed from activated cytoplasts, in contrast, exhibited a complete nuclear envelope at all...... time intervals after fusion. In the two-cell stage nuclear transfer embryo, the originally reticulated nucleolus of the donor blastomere had changed into a typical nucleolar precursor body consisting of a homogeneous fibrillar structure. A primary vacuole appeared in the four-cell stage nuclear...

  6. Progesterone elevation on the day of human chorionic gonadotropin administration adversely affects the outcome of IVF with transferred embryos at different developmental stages.

    Science.gov (United States)

    Huang, Yan; Wang, En-Yin; Du, Qing-Yun; Xiong, Yu-Jing; Guo, Xiao-Yi; Yu, Yi-Ping; Sun, Ying-Pu

    2015-08-04

    The effect of progesterone elevation (PE) on the day of human chorionic gonadotropin (hCG) administration on the pregnancy outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles is a matter of ongoing debate. The replacement of cleavage-stage embryos with blastocyst-stage embryos for transfer was proposed to avoid the possible impairment of PE in fresh cycles. This study aimed to assess the association between PE on the day of human chorionic gonadotropin (hCG) administration and clinical pregnancy rates (CPRs) in IVF/ICSI cycles with embryos transferred at different developmental stages (cleavage and blastocyst). Moreover, a secondary aim was to determine the thresholds at which PE has a detrimental effect on CPRs. This single-center retrospective cohort study included more than 10,000 patients undergoing day 3 cleavage-stage embryo transfer (ET) and 1146 patients undergoing day 5 blastocyst-stage embryo transfer (ET) using gonadotropin and GnRH agonist for controlled ovarian stimulation. Serum PE was inversely associated with CPRs in both cleavage- and blastocyst-stage ET cycles. In the day 3 ET cycles, CPRs (progesterone levels < 0.5 ng/ml, 49.2 %) significantly declined when the progesterone concentration reached 1.0 ng/ml (45.5 %) and decreased further when the progesterone concentration increased to 1.5 ng/ml (36.2 %). In the day 5 blastocyst-stage ET cycles, patients with serum progesterone levels ≥1.75 ng/ml had significantly lower CPRs (31.3 % VS. 41.4 %, p < 0.001) compared to patients with serum progesterone levels <1.75 ng/ml. The negative association of PE with CPRs was noted in both ET groups, even after adjusting for confounders. Furthermore, the developmental stage of the transferred embryos was not linked to the effect of PE on CPRs because the interaction between the developmental stage of the transferred embryos and PE was not significant. PE on the day of hCG administration is associated with

  7. Optimizing the number of cleavage stage embryos to transfer on day 3 in women 38 years of age and older: a Society for Assisted Reproductive Technology database study.

    Science.gov (United States)

    Stern, Judy E; Goldman, Marlene B; Hatasaka, Harry; MacKenzie, Todd A; Surrey, Eric S; Racowsky, Catherine

    2009-03-01

    To determine the optimal number of day 3 embryos to transfer in women >or=38 years by conducting an evidence-based evaluation. Retrospective analysis of 2000-2004 national SART data. National writing group. A total of 36,103 day 3 embryo transfers in women >or=38 years undergoing their first assisted reproductive technology cycle. None. Logistic regression was used to model the probability of pregnancy, delivery, and multiple births (twin or high order) based on age- and cycle-specific parameters. Pregnancy rates, delivery rates, and multiple rates increased up to transfer of three embryos in 38-year-olds and four in 39-year-olds; beyond this number, only multiple rates increased. In women >or=40 years, delivery rates and multiple rates climbed steadily with increasing numbers transferred. Multivariate analysis confirmed the statistically significant effect of age, number of oocytes retrieved, and embryo cryopreservation on delivery and multiple rates. Maximum FSH level was not an independent predictor by multivariate analysis. Use of intracytoplasmic sperm injection was associated with lowered delivery rate. No more than three or four embryos should be transferred in 38- and 39-year-olds, respectively, whereas up to five embryos could be transferred in >or=40-year-olds. Numbers of embryos to transfer should be adjusted according to number of oocytes retrieved and availability of excess embryos for cryopreservation.

  8. Cryopreservation of biopsied cleavage stage human embryos.

    Science.gov (United States)

    Stachecki, James J; Cohen, Jacques; Munné, Santiago

    2005-12-01

    The aim was to develop a method to optimize cryopreservation of biopsied multi-celled human embryos. Human day 3 embryos that were donated to research, along with those found to be chromosomally abnormal after blastomere biopsy and fluorescence in-situ hyridization (FISH), were cryopreserved using a slow-freezing protocol in either standard embryo cryopreservation solution [embryo transfer freezing medium (ETFM), a conventional sodium-based medium] or CJ3 (a choline-based, sodium-free medium). After thawing, the number of intact cells was recorded and the previously biopsied embryos were re-analysed using FISH. Biopsied embryos had a lower proportion of intact blastomeres after cryopreservation as compared with intact embryos. However, a significantly (P < 0.05) higher proportion of blastomeres from intact and biopsied embryos cryopreserved in CJ3 (84.1 and 80.1% respectively) survived after thaw than those in ETFM (73.6 and 50.5% respectively). The proportion of aneuploid and mosaic embryos was not statistically different between the two groups. In addition, the frequency of lost cells by aneuploid and mosaic embryos was similar. This study describes a new method that improves the survival of cryopreserved biopsied embryos, and shows that it may also be beneficial for the storage of intact human multi-celled embryos.

  9. Nucleolar remodeling in nuclear transfer embryos

    DEFF Research Database (Denmark)

    Laurincik, Jozef; Maddox-Hyttel, Poul

    2007-01-01

    nucleoli are not apparent until the 5th cell cycle, whereas in somatic cell nuclear transfer embryos the functional nucleoli emerge already during the 3rd cell cycle. Intergeneric reconstructed embryos produced by the fusion of bovine differentiated somatic cell to a nonactivated ovine cytoplast fail...... the developmental potential of embryos originating from varied nuclear transfer protocols. In bovine in vivo developed embryos, functional ribosome-synthesizing nucleoli become structurally distinct toward the end of the 4th post-fertilization cell cycle. In embryonic cell nuclear transfer embryos, fully developed...... is completed toward the end of the 4th cell cycle. A substantial proportion of bovine embryos produced by nuclear transfer of embryonic or somatic cells to bovine ooplasts display aberrations in protein localization in one or more blastomers. This information is indicative of underlying aberrations in genomic...

  10. Embryo transfer in domestic South American camelids.

    Science.gov (United States)

    Sumar, Julio B

    2013-01-10

    Intraspecific and interspecific embryo transfer in domestic South American camelids is developing into a well-established technique. Reports reveal many benefits of using reproductive biotechnologies to allow rapid propagation of alpacas and llamas of high genetic merit (e.g., high fiber quality, preserve color variation). The objective of this review is to provide up-to-date information about embryo transfer in domestic South American camelids. Specific information is provided on criteria for male selection, donor and recipient synchronization, the practice of single- vs. super-ovulation protocols, embryo recovery and transfer techniques, advances in cryopreservation of embryos, results of intra- and inter-specific transfer, and the future of the embryo transfer in domestic South American camelids. Copyright © 2012. Published by Elsevier B.V.

  11. Multiple-embryo transfer for studying very early maternal-embryo interactions in cattle.

    Science.gov (United States)

    Gómez, E; Muñoz, M

    2015-08-01

    In the present paper, we highlight the need to study very early maternal-embryo interactions and discuss how these interactions can be addressed. Bovine species normally carry one or, less frequently, two embryos to term; there are very rare cases of triplets or higher-order multiple pregnancies in which all the offspring are born alive. Multiple-embryo transfer (MET) in cattle allows for the detection of endometrial responses in scenarios where single-embryo transfer would not. Although MET is non-physiological, the present study shows that at the very early embryonic stages, a uterus carrying zona-enclosed embryos does not exhibit non-physiological reactions. On the contrary, MET should be considered the sum of multiple individual effects triggered by developing embryos. We provide arguments to support our hypothesis that describe a rationale for current work with MET, and we discuss alternative hypotheses. Using cattle as a model, we describe how technical approaches to analyzing zona-enclosed early embryo-maternal interactions (i.e., transcriptomics, proteomics, and endometrial cell culture) can help identify molecular changes that may be difficult to observe when only a single embryo is present. We conclude that MET can be used for studying very early maternal-embryo interactions in vivo in monotocous species. Free Spanish abstract: A Spanish translation of this abstract is freely available at http://www.reproduction-online.org/content/150/2/R35/suppl/DC1. © 2015 Society for Reproduction and Fertility.

  12. Embryo transfer using cryopreserved Boer goat blastocysts ...

    African Journals Online (AJOL)

    The aim of this trial was to evaluate the effect of embryo cryopreservation techniques on the survivability of embryos and fertility following transfer to Boer goat does. The oestrous cycles of 27 mature recipients Boer goat does were synchronised using controlled internal drug release dispensers (CIDR's) for 16 days. At CIDR ...

  13. Nucleolar remodeling in nuclear transfer embryos

    DEFF Research Database (Denmark)

    Laurincik, Jozef; Maddox-Hyttel, Poul

    2007-01-01

    the developmental potential of embryos originating from varied nuclear transfer protocols. In bovine in vivo developed embryos, functional ribosome-synthesizing nucleoli become structurally distinct toward the end of the 4th post-fertilization cell cycle. In embryonic cell nuclear transfer embryos, fully developed...... to develop fully functional nucleoli. In bovine in vivo developed embryos, a range of important nucleolar proteins (e.g., topoisomerase I, upstream binding factor and RNA polymerase I, fibrillarin, nucleophosmin and nucleolin) become localized to the nucleolar anlage over several cell cycles. This relocation......Transcription of the ribosomal RNA (rRNA) genes occurs in the nucleolus and results in ribosome biogenesis. The rRNA gene activation and the associated nucleolus formation may be used as a marker for the activation of the embryonic genome in mammalian embryos and, thus serve to evaluate...

  14. Debating Elective Single Embryo Transfer after in vitro Fertilization ...

    African Journals Online (AJOL)

    transfer (eSET) to reduce the risk of multiple gestation and. Debating Elective Single Embryo Transfer ... the American Society for Reproductive Medicine, elective SET is defined as “the transfer of a single embryo at ... or should they transfer just one embryo, which reduces the risk of complications and is in line with current ...

  15. Live embryo imaging to follow cell cycle and chromosomes stability after nuclear transfer.

    Science.gov (United States)

    Balbach, Sebastian T; Boiani, Michele

    2015-01-01

    Nuclear transfer (NT) into mouse oocytes yields a transcriptionally and functionally heterogeneous population of cloned embryos. Most studies of NT embryos consider only embryos at predefined key stages (e.g., morula or blastocyst), that is, after the bulk of reprogramming has taken place. These retrospective approaches are of limited use to elucidate mechanisms of reprogramming and to predict developmental success. Observing cloned embryo development using live embryo cinematography has the potential to reveal otherwise undetectable embryo features. However, light exposure necessary for live cell cinematography is highly toxic to cloned embryos. Here we describe a protocol for combined bright-field and fluorescence live-cell imaging of histone H2b-GFP expressing mouse embryos, to record cell divisions up to the blastocyst stage. This protocol, which can be adapted to observe other reporters such as Oct4-GFP or Nanog-GFP, allowed us to quantitatively analyze cleavage kinetics of cloned embryos.

  16. Embryo aggregation does not improve the development of interspecies somatic cell nuclear transfer embryos in the horse.

    Science.gov (United States)

    Gambini, Andrés; De Stéfano, Adrián; Jarazo, Javier; Buemo, Carla; Karlanian, Florencia; Salamone, Daniel Felipe

    2016-09-01

    The low efficiency of interspecies somatic cell nuclear transfer (iSCNT) makes it necessary to investigate new strategies to improve embryonic developmental competence. Embryo aggregation has been successfully applied to improve cloning efficiency in mammals, but it remains unclear whether it could also be beneficial for iSCNT. In this study, we first compared the effect of embryo aggregation over in vitro development and blastocyst quality of porcine, bovine, and feline zona-free (ZF) parthenogenetic (PA) embryos to test the effects of embryo aggregation on species that were later used as enucleated oocytes donors in our iSCNT study. We then assessed whether embryo aggregation could improve the in vitro development of ZF equine iSCNT embryos after reconstruction with porcine, bovine, and feline ooplasm. Bovine- and porcine-aggregated PA blastocysts had significantly larger diameters compared with nonaggregated embryos. On the other hand, feline- and bovine-aggregated PA embryos had higher blastocyst cell number. Embryo aggregation of equine-equine SCNT was found to be beneficial for embryo development as we have previously reported, but the aggregation of three ZF reconstructed embryos did not improve embryo developmental rates on iSCNT. In vitro embryo development of nonaggregated iSCNT was predominantly arrested around the stage when transcriptional activation of the embryonic genome is reported to start on the embryo of the donor species. Nevertheless, independent of embryo aggregation, equine blastocyst-like structures could be obtained in our study using domestic feline-enucleated oocytes. Taken together, these results reported that embryo aggregation enhance in vitro PA embryo development and embryo quality but effects vary depending on the species. Embryo aggregation also improves, as expected, the in vitro embryo development of equine-equine SCNT embryos; however, we did not observe positive effects on equine iSCNT embryo development. Among oocytes

  17. To delay or not to delay a frozen embryo transfer after a failed fresh embryo transfer attempt?

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    Santos-Ribeiro, Samuel; Siffain, Johannie; Polyzos, Nikolaos P; van de Vijver, Arne; van Landuyt, Lisbet; Stoop, Dominic; Tournaye, Herman; Blockeel, Christophe

    2016-05-01

    To evaluate if increasing the interval between a failed fresh embryo transfer and a subsequent frozen embryo transfer (FET) cycle has any effect on clinical pregnancy rates (CPRs). Retrospective cohort study. University-based tertiary referral center. Women who underwent at least one FET after ovarian stimulation for in vitro fertilization (IVF) and a failed fresh embryo transfer attempt from January 2010 to November 2014. We divided our sample according to the "timing" of the first FET (TF-FET), defined by the interval between oocyte retrieval and the FET cycle start date. The start of the FET was classified as either immediate (≤22 days after oocyte retrieval) or delayed (>22 days after oocyte retrieval). None. CPR after the first FET. A total of 1,183 FET cycles (performed in 1,087 women) were included in our study. No significant differences were found between the immediate and delayed FET groups regarding age, number of oocytes retrieved, number of good-quality embryos produced, embryo developmental stage at FET, and number of frozen embryos transferred. Most importantly, the CPRs of the first FET did not differ significantly according to the TF-FET (32.5% after immediate FET vs. 31.7% after delayed FET), even after adjusting for potential confounding with the use of multivariable logistic regression. FETs performed immediately after fresh IVF cycles had CPRs similar to those postponed to a later time. Therefore, deferring FETs may unnecessarily prolong time to pregnancy. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  18. Comparison between Conventional Blind Embryo Transfer and Embryo Transfer Based on Previously Measured Uterine Length

    Directory of Open Access Journals (Sweden)

    Nasrin Saharkhiz

    2014-11-01

    Full Text Available Background: Embryo transfer (ET is one of the most important steps in assisted reproductive technology (ART cycles and affected by many factors namely the depth of embryo deposition in uterus. In this study, the outcomes of intracytoplasmic sperm injection (ICSI cycles after blind embryo transfer and embryo transfer based on previously measured uterine length using vaginal ultrasound were compared. Materials and Methods: This prospective randomised clinical trial included one hundred and forty non-donor fresh embryo transfers during January 2010 to June 2011. In group I, ET was performed using conventional (blind method at 5-6cm from the external os, and in group II, ET was done at a depth of 1-1.5 cm from the uterine fundus based on previously measured uterine length using vaginal sonography. Appropriate statistical analysis was performed using Student’s t test and Chi-square or Fisher’s exact test. The software that we used was PASW statistics version 18. A p value <0.05 was considered statistically significant. Results: Chemical pregnancy rate was 28.7% in group I and 42.1% in group II, while the difference was not statistically significant (p=0.105. Clinical pregnancy, ongoing pregnancy and implantation rates for group I were 21.2%, 17.7%, and 12.8%, while for group II were 33.9%, 33.9%, and 22.1, respectively. In group I and group II, abortion rates were 34.7% and 0%, respectively, indicating a statistically significant difference (p<0.005. No ectopic pregnancy occurred in two groups. Conclusion: The use of uterine length measurement during treatment cycle in order to place embryos at depth of 1-1.5cm from fundus significantly increases clinical and ongoing pregnancy and implantation rates, while leads to a decrease in abortion rate (Registration Number: IRCT2014032512494N1.

  19. Development of sheep androgenetic embryos is boosted following transfer of male pronuclei into androgenetic hemizygotes.

    Science.gov (United States)

    Matsukawa, Kazutsugu; Turco, Margherita Yayoi; Scapolo, Pier Augusto; Reynolds, Lawrence; Ptak, Grazyna; Loi, Pasqualino

    2007-01-01

    Androgenetic embryos are useful model for investigating the contribution of the paternal genome to embryonic development. Little work has been done with androgenetic embryo production in domestic animals. The aim of this study was the production of diploid androgenetic sheep embryos. In vitro matured sheep oocytes were enucleated and fertilized in vitro; parthenogenetic and normally fertilized embryos were also produced as a control. Fifteen hours after in vitro fertilization (IVF), presumptive zygotes were centrifuged and scored for the number of pronucleus. IVF, parthenogenetic, and androgenetic embryos (haploid, diploid, and triploid) were cultured in SOFaa medium with bovine serum albumin (BSA). The proportion of oocytes with polyspermic fertilization increased linearly with increasing sperm concentration. After IVF, there was no significant difference in early cleavage and morula formation rates between the groups, while there was a significant difference on blastocyst development between IVF, parthenogenetic, and androgenetic embryos, the last ones displaying poor developmental potential (IVF, parthenogenetic, and haploid, diploid, and triploid androgenetic embryos: 43%, 38%, 0%, 2%, and 2%, respectively). In order to boost androgenetic embryonic development, we produced diploid androgenetic embryos through pronuclear transfer. Single pronuclei were aspirated with a bevelled pipette from haploid or diploid embryos and transferred into the perivitelline space of other haploid embryos, and the zygotes were reconstructed by electrofusion. Fusion rates approached 100%. Pronuclear transfer significantly increased blastocyst development (IVF, parthenogenetic, androgenetic: Diploid into Haploid, and Haploid into Haploid: 42%, 42%, 19%, and 3%, respectively); intriguingly, the Haploid + Diploid group showed the highest development to blastocyst stage. The main findings of our study are: (1) sheep androgenetic embryos display poor developmental ability compared with

  20. The effect of the number of transferred embryos, the interval between nuclear transfer and embryo transfer, and the transfer pattern on pig cloning efficiency.

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    Rim, Chol Ho; Fu, Zhixin; Bao, Lei; Chen, Haide; Zhang, Dan; Luo, Qiong; Ri, Hak Chol; Huang, Hefeng; Luan, Zhidong; Zhang, Yan; Cui, Chun; Xiao, Lei; Jong, Ui Myong

    2013-12-01

    To improve the efficiency of producing cloned pigs, we investigated the influence of the number of transferred embryos, the culturing interval between nuclear transfer (NT) and embryo transfer, and the transfer pattern (single oviduct or double oviduct) on cloning efficiency. The results demonstrated that transfer of either 150-200 or more than 200NT embryos compared to transfer of 100-150 embryos resulted in a significantly higher pregnancy rate (48 ± 16, 50 ± 16 vs. 29 ± 5%, pcloning efficiency is achieved by adjusting the number and in vitro culture time of reconstructed embryos as well as the embryo transfer pattern. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Embryo transfer and related technologies in sheep reproduction

    OpenAIRE

    Loi, Pasqualino; Ptak, Grazyna; Dattena, Maria; Ledda, Sergio; Naitana, Salvatore; Cappai, Pietro

    1998-01-01

    This paper reviews the status of embryo transfer and the major technologies applied to preimplantation of embryos in sheep. Embryo production from superovulated ewes is hindered by an unpredictable response to hormonal treatment. Progress in this area should be expected by an appropriated control of follicular development with gonadotropin-releasing hormone (GnRH) agonist or antagonist prior to gonadotrophin administration. Simple protocols for the cryopreservation of sheep embryos by vitrifi...

  2. Successful pregnancy following trans-myometrial embryo transfer ...

    African Journals Online (AJOL)

    ... factor infertility (poor motility). Routine mock embryo transfer indicated severe cervical stenosis which was confirmed at hysteroscopy. She subsequently had trans-myometrial embryo transfer. Blood pregnancy test at 2 weeks was positive and interval transvaginal ultrasound confirmed 2 viable intrauterine foetal poles.

  3. Factors affecting conception rates in cattle following embryo transfer ...

    African Journals Online (AJOL)

    Embryo Transfer Technology (ETT) plays an important role in improving productivity of dairy cattle (Bos indicus). Embryo Transfer Technology allows top quality female livestock to improve a herd or flock in much the same way that artificial insemination has allowed greater use of superior sires. The technology hastens ...

  4. A new direct transfer protocol for cryopreserved IVF embryos.

    Science.gov (United States)

    Sanches, Bruno Valente; Lunardelli, Paula Alvares; Tannura, Juliana Hayashi; Cardoso, Bruna Lopes; Pereira, Marcos Henrique Colombo; Gaitkoski, Douglas; Basso, Andrea Cristina; Arnold, Daniel Robert; Seneda, Marcelo Marcondes

    2016-04-01

    The global demand for in vitro-produced (IVP) embryos of determined sex has greatly increased over the last decade. Efficient protocols for the direct transfer of IVP embryos are lacking. This study aimed to compare the pregnancy rates for fresh, vitrified, or frozen/directly transferred IVP dairy cow embryos. Oocytes (n = 3171) recovered by ovum pickup (n = 112) from Girolando (Holstein-Gir) females (n = 36) were selected and submitted to IVM for 24 hours at 38.5 °C with 5% CO2 in air with saturated humidity. In vitro fertilization was performed with the thawed, sexed semen from 5 Holstein bulls. After IVF, presumptive zygotes were denuded and cultured for 7 days under the same IVM and IVF conditions of temperature and humidity, except with 5% CO2 and 5% O2. Grade I blastocysts were randomly assigned for either the transferred fresh, vitrified/thawing, or frozen/directly embryo transfer into previously synchronized recipient females. Conception rates were analyzed by binomial logistic regression, and a probability level of P < 0.05 was considered significant. The conception rates were 51.35 ± 1.87% (133/259) for the fresh embryos, 35.89 ± 3.87% (84/234) for the vitrified embryos, and 40.19 ± 4.65% (125/311) for the frozen directly transferred embryos. These data demonstrate that IVP embryos with sexed semen could be directly transferred into recipient cows with similar conception rates to vitrified embryos. The comparison found that the use of frozen embryos in direct transfer provides easier logistics and a more practical approach for the transfer of IVP embryos on dairy farms. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Multiple ovulation and embryo transfer (MOET in camels: An overview

    Directory of Open Access Journals (Sweden)

    Binoy S. Vettical

    2016-04-01

    Full Text Available Unlike in other domestic animal species like cattle, reproductive biotechnologies like Artificial Insemination (AI and Embryo Transfer (ET are not well developed and thus are not being used as routine breeding procedures in camels. One of the important objectives of this manuscript is to focus on analyzing the present status of Multiple Ovulation and Embryo Transfer (MOET in camels and its future perspectives. Camels are induced ovulators, thus require hormonal treatment to induce ovulation and control the follicular cycles, which is the main reason why protocols used in other domestic animal species cannot be directly used in this species. The review suggests that the best method for super stimulation of ovaries in camels is use of a combination of Equine Chorionic Gonadotropin (eCG and Follicle Stimulating Hormone (FSH at any stage after elimination of dominant follicle if any or at the early stage of the follicular wave and ovulation of the developed multiple follicles can be achieved by mating donors. The review highlights that a better pregnancy rate is achieved with recipients who ovulate 24 h after the donor.

  6. Pouch brooding marsupial frogs transfer nutrients to developing embryos.

    Science.gov (United States)

    Warne, Robin W; Catenazzi, Alessandro

    2016-10-01

    Marsupial frogs have a unique reproductive mode in which females carry eggs enclosed in a sealed dorsal brood pouch. While most anurans are considered to be oviparous with lecithotrophic eggs, the extensively vascularized membrane of the brood pouch in marsupial frogs suggests potential opportunities for nutrient transfer. We tested for matrotrophy in the live-bearing Gastrotheca excubitor (Hemiphractidae), through feeding insects labelled with a 13 C-fatty acid and a 15 N-amino acid to brooding marsupial frogs. We observed significant increases of δ 13 C and δ 15 N in both maternal pouch tissues and embryos, suggesting nutrient transfer. Embryo dry mass also increased with developmental stage, providing further direct evidence for matrotrophy. These results suggest that in addition to gas exchange, the vascularized brood pouch membrane of G. excubitor also enables maternal nutrient transfer. This finding revealed a suspected but untested trait in the evolution of parental care in marsupial frogs, in contrast to previous work on Gastrotheca species that release tadpoles, and suggests greater complexity in reproductive and provisioning modes than previously thought. © 2016 The Author(s).

  7. Factors that affect the reproductive efficiency of the recipient within a bovine embryo transfer program

    Directory of Open Access Journals (Sweden)

    Arturo Duica A.

    2007-12-01

    Full Text Available The embryo transfer is a biotechnological technique that allows increasing the descendant of animals with high genetic value. The positive results, represented in pregnancy after the application of this technique, are affected by some factors that are inherent to the donor, the embryo, the technique, and the recipients which receive a strange embryo in the uterus allowing pregnancy. This review describes some factors affecting the reproductive efficiency of the recipients of bovine embryos within a program of embryo transfer. Its important to evaluate the parameters in this kind of recipients, as race, age, physiological status, health status, weight, reproductive tract integrity and management, and also too monitoring the ovarian structures while the estrus synchronization, and within previous and posterior stages in embryo transfer procedure. Therefore an optimum follicular development will be determinant to corpus luteum formation which generates enough serum progesterone concentrations to offer a right uterine environment allowing the optimum embryo development. Controlling the factors that affect the efficiency of the embryo transfer, it will obtain an increasing of positive results represented in pregnancies and births of individuals come from animals with high genetic value.

  8. Biopsy of human morula-stage embryos: outcome of 215 IVF/ICSI cycles with PGS.

    Directory of Open Access Journals (Sweden)

    Elena E Zakharova

    Full Text Available Preimplantation genetic diagnosis (PGD is commonly performed on biopsies from 6-8-cell-stage embryos or blastocyst trophectoderm obtained on day 3 or 5, respectively. Day 4 human embryos at the morula stage were successfully biopsied. Biopsy was performed on 709 morulae from 215 ICSI cycles with preimplantation genetic screening (PGS, and 3-7 cells were obtained from each embryo. The most common vital aneuploidies (chromosomes X/Y, 21 were screened by fluorescence in situ hybridization (FISH. No aneuploidy was observed in 72.7% of embryos, 91% of those developed to blastocysts. Embryos were transferred on days 5-6. Clinical pregnancy was obtained in 32.8% of cases, and 60 babies were born. Patients who underwent ICSI/PGS treatment were compared with those who underwent standard ICSI treatment by examining the percentage of blastocysts, pregnancy rate, gestational length, birth height and weight. No significant differences in these parameters were observed between the groups. Day 4 biopsy procedure does not adversely affect embryo development in vitro or in vivo. The increased number of cells obtained by biopsy of morulae might facilitate diagnostic screening. There is enough time after biopsy to obtain PGD results for embryo transfer on day 5-6 in the current IVF cycle.

  9. ECTOPIC PREGNANCY AFTER SEQUENTIAL EMBRYO TRANSFER: REVIEW OF 22 CASES

    OpenAIRE

    Nadkarni Purnima K, Nadkarni Kishore, Singh Pooja P, Singh Prabhakar , Nadkarni Aditi A , Agarwal Neha R

    2015-01-01

    Objective: To assess the prevalence of ectopic pregnancy among women who conceived with assisted reproductive technology and to see if there is increased risk after sequential embryo transfer. Methods: The ectopic pregnancy rate for ART pregnancies was calculated among women who conceived and had ectopic pregnancy after ICSI followed by Sequential embryo transfer from an ART centre. Variation in ectopic risk by patient and ART treatment factors was assessed including Sequential transfer, risk...

  10. Fresh versus frozen embryo transfers in assisted reproduction.

    Science.gov (United States)

    Wong, Kai Mee; van Wely, Madelon; Mol, Femke; Repping, Sjoerd; Mastenbroek, Sebastiaan

    2017-03-28

    In general, in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) implies a single fresh and one or more frozen-thawed embryo transfers. Alternatively, the 'freeze-all' strategy implies transfer of frozen-thawed embryos only, with no fresh embryo transfers. In practice, both strategies can vary technically including differences in freezing techniques and timing of transfer of cryopreservation, that is vitrification versus slow freezing, freezing of two pro-nucleate (2pn) versus cleavage-stage embryos versus blastocysts, and transfer of cleavage-stage embryos versus blastocysts.In the freeze-all strategy, embryo transfers are disengaged from ovarian stimulation in the initial treatment cycle. This could avoid a negative effect of ovarian hyperstimulation on the endometrium and thereby improve embryo implantation. It could also reduce the risk of ovarian hyperstimulation syndrome (OHSS) in the ovarian stimulation cycle by avoiding a pregnancy.We compared the benefits and risks of the two treatment strategies. To evaluate the effectiveness and safety of the freeze-all strategy compared to the conventional IVF/ICSI strategy in women undergoing assisted reproductive technology. We searched the Cochrane Gynaecology and Fertility Group Trials Register, the Cochrane Central Register of Studies (CRSO), MEDLINE, Embase, PsycINFO, CINAHL, and two registers of ongoing trials in November 2016 together with reference checking and contact with study authors and experts in the field to identify additional studies. We included randomised clinical trials comparing a freeze-all strategy with a conventional IVF/ICSI strategy which includes fresh transfer of embryos in women undergoing IVF or ICSI treatment. We used standard methodological procedures recommended by Cochrane. The primary review outcomes were cumulative live birth and OHSS. Secondary outcomes included other adverse effects (miscarriage rate). We included four randomised clinical trials analysing a total

  11. Patients' Preference for Number of Embryos Transferred During IVF ...

    African Journals Online (AJOL)

    Background: The Human Fertilization and Embryology Authority is considering limiting the number of embryos that can be transferred to single embryo per cycle as has been done in several European countries, with the aim of reducing the rate of multiple pregnancies and its attendant complications following in vitro ...

  12. Production, Preservation, and Transfer of South American Camelid Embryos

    Directory of Open Access Journals (Sweden)

    Virginia L. Trasorras

    2017-11-01

    Full Text Available The current review summarizes progress in the field of in vitro and in vivo production of South American Camelid embryos. Both methods require ovarian superstimulation (with FSH and eCG to obtain multiple ovulations (in vivo embryo production or to induce follicle growth for oocyte collection (in vitro embryo production. Moreover, superstimulation entails prior administration of hormones that inhibit follicular growth (progesterone, progestagens, and estrogens. Cumulus-oocyte complexes obtained must mature in vivo (buserelin administration or in vitro to then be subjected to in vitro fertilization or intracytoplasmic sperm injection. All these techniques also require morphologically normal, motile spermatozoa to achieve fertilization. Methods used to decrease semen viscosity and to select the best spermatozoa (Percoll®; Androcoll-ETM are described. Additionally, nuclear transfer or cloning has been applied in llamas. Up to now, embryo deep-freezing and vitrification have progressed slowly but are at the height of development. Embryos that are obtained by any of these techniques, either in vivo or in vitro, need to be transferred to synchronized recipient females. The best results are achieved after transfer to the left uterine horn with an ipsilateral ovulation. No live offspring have been obtained after the transfer of cryopreserved embryos. Applying reproductive biotechnologies, such as those described, will permit the expansion of genetically selected animals in the population and also that of wild camelid species, vicunas, and guanacos, whose embryos could then be transferred to the uterus of domestic species.

  13. Transferring embryos with indeterminate PGD results: the ethical implications.

    Science.gov (United States)

    Insogna, Iris G; Ginsburg, Elizabeth

    2016-01-01

    In vitro fertilization (IVF) and pre-implantation genetic diagnosis (PGD) are becoming increasingly common techniques to select embryos that are unaffected by a known genetic disorder. Though IVF-PGD has high success rates, 7.5 % of blastocysts have inconclusive results after testing. A recent case involving a known BRCA-1 carrier was brought before our Assisted Reproductive Technology Ethics Committee in order to gain a better appreciation for the ethical implications surrounding the transfer of embryos with indeterminate testing. Thirty-nine year old G0 BRCA-1 carrier requiring IVF for male factor infertility. The couple elected for PGD to select against BRCA-1 gene carrier embryos. However, several embryos were returned with inconclusive results. The couple wished to proceed with the transfer of embryos with an unknown carrier status. The case was presented before our Assisted Reproductive Technology Ethics Committee. Many considerations were explored, including the physician's duty to protect patient autonomy, the physician's duty to act in the best interest of the future child, and the physician's duty towards society. Transferring both embryos with unknown carrier status and known-carrier status was debated. Ultimately, the transfer of inconclusive embryos was felt to be ethically permissible in most cases if patients had been adequately counseled. However, the re-biopsy of embryos with inconclusive testing results was encouraged. The transfer of known-carrier embryos was felt to be unethical for certain disease-states, depending on the severity of illness and timing of disease onset. We strongly encourage physicians to create an action plan in advance with their patients, prior to testing, in the event that embryos are returned with inconclusive PGD results. The committee's decision, though helpful in guiding practice, should not overshadow the individual physician-patient relationship, and the need for thorough counseling.

  14. Can Chlamydia abortus be transmitted by embryo transfer in goats?

    Science.gov (United States)

    Oseikria, M; Pellerin, J L; Rodolakis, A; Vorimore, F; Laroucau, K; Bruyas, J F; Roux, C; Michaud, S; Larrat, M; Fieni, F

    2016-10-01

    The objectives of this study were to determine (i) whether Chlamydia abortus would adhere to or penetrate the intact zona pellucida (ZP-intact) of early in vivo-derived caprine embryos, after in vitro infection; and (ii) the efficacy of the International Embryo Transfer Society (IETS) washing protocol for bovine embryos. Fifty-two ZP-intact embryos (8-16 cells), obtained from 14 donors were used in this experiment. The embryos were randomly divided into 12 batches. Nine batches (ZP-intact) of five embryos were incubated in a medium containing 4 × 10(7)Chlamydia/mL of AB7 strain. After incubation for 18 hours at 37 °C in an atmosphere of 5% CO2, the embryos were washed in batches in 10 successive baths of a phosphate buffer saline and 5% fetal calf serum solution in accordance with IETS guidelines. In parallel, three batches of ZP-intact embryos were used as controls by being subjected to similar procedures but without exposure to C. abortus. The 10 wash baths were collected separately and centrifuged for 1 hour at 13,000 × g. The washed embryos and the pellets of the 10 centrifuged wash baths were frozen at -20 °C before examination for evidence of C. abortus using polymerase chain reaction. C. abortus DNA was found in all of the infected batches of ZP-intact embryos (9/9) after 10 successive washes. It was also detected in the 10th wash fluid for seven batches of embryos, whereas for the two other batches, the last positive wash bath was the eighth and the ninth, respectively. In contrast, none of the embryos or their washing fluids in the control batches were DNA positive. These results report that C. abortus adheres to and/or penetrates the ZP of in vivo caprine embryos after in vitro infection, and that the standard washing protocol recommended by the IETS for bovine embryos, failed to remove it. The persistence of these bacteria after washing makes the embryo a potential means of transmission of the bacterium during embryo transfer from

  15. Embryo transfer day does not affect the initial maternal serum β-hCG levels: A retrospective cohort study.

    Science.gov (United States)

    Dahiya, Mona; Rupani, Karishma; Yu, Su Ling; Fook-Chong, Stephanie M C; Siew Fui, Diana Chia; Rajesh, Hemashree

    2017-05-01

    The aim of this study is to compare the serum β-hCG values post transfer of a cleavage stage embryo versus a blastocyst stage embryo at equal time intervals post oocyte retrieval (OR) in clinically pregnant patients, and to ascertain a β-hCG value to predict pregnancy outcomes. This is a retrospective cohort study of 560 women with clinical pregnancy who underwent an embryo transfer performed at either the cleavage stage or the blastocyst stage of embryo development between January 2003 and June 2014 at the Center for Assisted Reproduction (CARE), Singapore General Hospital. The serum β-hCG level was measured on day 17 post OR. The β-hCG values were not significantly different in the cleavage stage versus the blastocyst stage embryos (mean±SD: 387±486IU/L D3 vs. 352±268IU/L D5, p=0.96, median value 297 in both groups). Our study suggests that the initial maternal serum β-hCG values were not affected by the day of transfer of the embryos since assessing the β-hCG at equivalent points after transfer should not lead to a significant difference assuming the progress and development of the embryos occurred as expected. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Timing of oocyte retrieval and embryo transfer with pregnancy duration.

    Science.gov (United States)

    Beydoun, Hind A; Ugwu, Bethrand; Indika, Sathish; Stadtmauer, Laurel; Bocca, Silvina; Oehninger, Sergio

    2011-11-01

    Recent studies have suggested that assisted reproductive technology (ART) may be associated with a shorter pregnancy duration, possibly due to various aspects of the ART procedure. The purpose of this study was to examine whether pregnancy duration is affected by timing of oocyte retrieval and embryo transfer with respect to the first day of the last menstrual period (LMP) among pregnancies achieved through in vitro fertilization with or without intracytoplasmic sperm injection. A retrospective study was conducted at an academic center in Norfolk, Virginia, with analyses based on 294 ART cycles. Median and interquartile range for pregnancy duration was estimated at 38.2 ± 3.4 weeks. Similarly, median and interquartile ranges for days between LMP and day of oocyte retrieval (27.0 ± 2.0) and between LMP and embryo transfer (29.8 ± 2.2) differed significantly from the standard of 14 days. Timing of oocyte retrieval and embryo transfer with respect to LMP were accelerated among multiple compared with single gestations. For single gestations, pregnancy duration was positively associated with time duration between LMP and embryo transfer (β=0.14, p=0.036). The number of days between oocyte retrieval and embryo transfer was marginally associated with a shorter pregnancy duration in women with multiple gestations (β=3.70, p=0.083). Controlling for patient characteristics, timing of oocyte retrieval and embryo transfer were not significantly associated with pregnancy duration. With few exceptions, timing of oocyte retrieval or embryo transfer did not affect pregnancy duration among ART-conceived live births.

  17. Cost-effectiveness of single versus double embryo transfer in IVF in relation to female age

    NARCIS (Netherlands)

    van Loendersloot, Laura L.; Moolenaar, Lobke M.; van Wely, Madelon; Repping, Sjoerd; Bossuyt, Patrick M.; Hompes, Peter G. A.; van der Veen, Fulco; Mol, Ben Willem J.

    2017-01-01

    Objective: To evaluate the cost-effectiveness of single embryo transfer followed by an additional frozen thawed single embryo transfer, if more embryos are available, as compared to double embryo transfer in relation to female age. Study design: We used a decision tree model to evaluate the costs

  18. ECTOPIC PREGNANCY AFTER SEQUENTIAL EMBRYO TRANSFER: REVIEW OF 22 CASES

    Directory of Open Access Journals (Sweden)

    Nadkarni Purnima K, Nadkarni Kishore, Singh Pooja P, Singh Prabhakar , Nadkarni Aditi A , Agarwal Neha R

    2015-07-01

    Full Text Available Objective: To assess the prevalence of ectopic pregnancy among women who conceived with assisted reproductive technology and to see if there is increased risk after sequential embryo transfer. Methods: The ectopic pregnancy rate for ART pregnancies was calculated among women who conceived and had ectopic pregnancy after ICSI followed by Sequential embryo transfer from an ART centre. Variation in ectopic risk by patient and ART treatment factors was assessed including Sequential transfer, risk factor for ectopic pregnancy, tubal infertility and previous ectopic pregnancy. Results: Of 1960 women, who underwent ICSI, 1047(53.41% had positive pregnancy, 22(2.1% women had an ectopic pregnancy out of which 2(9% had heterotrophic pregnancy. The mean age was 30.4±4.33. Tubal factor contributed to 31.81% cases and 27.27% had previous history of ectopic pregnancy. Sequential transfer was done in all the patients with more than one embryo transferred. There was no significant increase in the ectopic pregnancy when Sequential embryo transfer was done. Conclusion: Tubal factor infertility and history of previous ectopic pregnancy contributed to risk factor for ectopic pregnancy in our study. Sequential transfer did not increase the risk of ectopic pregnancy.

  19. RECYCLING OF CATHETER FOR EMBRYO RECOVERY: A TOOL FOR COSTS REDUCTION IN EQUINE EMBRYO TRANSFER

    Directory of Open Access Journals (Sweden)

    Alberto Lopes Gusmao

    2010-06-01

    Full Text Available The embryo transfer is becoming a widespread practice.Most embryos are collected from spontaneous single ovulatingmares and result in 50% of embryo recovery, increasing the costsof production. To illustrate, the price of a catheter for embryosrecovery range from US$ 194.00 to US$ 250.00 (R$ 350.00 to R$450.00. Therefore, the aim of this work was to verify if catheterwith damaged balloon can be recuperated and reused withoutaltering its efficiency. For this study, two groups were used: acontrol group (GI, n=10, on which the nonsurgical recovery of theembryos of mares was performed with the catheter with originalballoon; and another group (GII, n=20, in which a restored catheterwas utilized. The mares of GI had an embryo recovery rate of60%, and GII mares had an embryo recovery rate of 55%. Therewas not statistical difference between groups I and II (P>0.05.Considering that the material used to restore the catheter costsUS$16.66 (R$30.00, this data show that the recuperation of thecatheters for embryo recovery in mares may reduce costs withoutcompromising the rates of embryo recovery.

  20. Production of rhesus monkey cloned embryos expressing monomeric red fluorescent protein by interspecies somatic cell nuclear transfer

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Hai-Ying; Kang, Jin-Dan; Li, Suo; Jin, Jun-Xue; Hong, Yu; Jin, Long; Guo, Qing; Gao, Qing-Shan; Yan, Chang-Guo; Yin, Xi-Jun, E-mail: yinxj33@msn.com

    2014-02-21

    Highlights: • Rhesus monkey cells were electroporated with a plasmid containing mRFP1, and an mRFP1-expressing cell line was generated. • For the first time, mRFP1-expressing rhesus monkey cells were used as donor cells for iSCNT. • The effect of VPA on the development of embryos cloned using iSCNT was determined. - Abstract: Interspecies somatic cell nuclear transfer (iSCNT) is a promising method to clone endangered animals from which oocytes are difficult to obtain. Monomeric red fluorescent protein 1 (mRFP1) is an excellent selection marker for transgenically modified cloned embryos during somatic cell nuclear transfer (SCNT). In this study, mRFP-expressing rhesus monkey cells or porcine cells were transferred into enucleated porcine oocytes to generate iSCNT and SCNT embryos, respectively. The development of these embryos was studied in vitro. The percentage of embryos that underwent cleavage did not significantly differ between iSCNT and SCNT embryos (P > 0.05; 71.53% vs. 80.30%). However, significantly fewer iSCNT embryos than SCNT embryos reached the blastocyst stage (2.04% vs. 10.19%, P < 0.05). Valproic acid was used in an attempt to increase the percentage of iSCNT embryos that developed to the blastocyst stage. However, the percentages of embryos that underwent cleavage and reached the blastocyst stage were similar between untreated iSCNT embryos and iSCNT embryos treated with 2 mM valproic acid for 24 h (72.12% vs. 70.83% and 2.67% vs. 2.35%, respectively). These data suggest that porcine-rhesus monkey interspecies embryos can be generated that efficiently express mRFP1. However, a significantly lower proportion of iSCNT embryos than SCNT embryos reach the blastocyst stage. Valproic acid does not increase the percentage of porcine-rhesus monkey iSCNT embryos that reach the blastocyst stage. The mechanisms underling nuclear reprogramming and epigenetic modifications in iSCNT need to be investigated further.

  1. In vitro development of OPU-derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen-thawed embryos after transfer.

    Science.gov (United States)

    Isobe, Tomohiro; Ikebata, Yoshihisa; Do, Lanh Thi Kim; Tanihara, Fuminori; Taniguchi, Masayasu; Otoi, Takeshige

    2015-07-01

    The optimization of single-embryo culture conditions is very important, particularly in the in vitro production of bovine embryos using the ovum pick-up (OPU) procedure. The purpose of this study was to examine the development of embryos derived from oocytes obtained by OPU that were cultured either individually or in groups in medium supplemented with or without sericin and to investigate the viability of the frozen-thawed embryos after a direct transfer. When two-cell-stage embryos were cultured either individually or in groups for 7 days in CR1aa medium supplemented with or without 0.5% sericin, the rates of development to blastocysts and freezable blastocysts were significantly lower for the embryos cultured individually without sericin than for the embryos cultured in groups with or without sericin. Moreover, the rate of development to freezable blastocysts of the embryos cultured individually with sericin was significantly higher than that of the embryos cultured without sericin. When the frozen-thawed embryos were transferred directly to recipients, the rates of pregnancy, abortion, stillbirth and normal calving in the recipients were similar among the groups, irrespective of the culture conditions and sericin supplementation. Our findings indicate that supplementation with sericin during embryo culture improves the quality of the embryos cultured individually but not the viability of the frozen-thawed embryos after transfer to recipients. © 2014 Japanese Society of Animal Science.

  2. Factors affecting conception rates in cattle following embryo transfer

    African Journals Online (AJOL)

    ACSS

    production, sale of breeding stock, among others. New technologies can help achieve increased productivity, but need to be transferred to producers to cause impact. (Ehui and Shapiro, 2009; Yang and. Honaramooz, 2010). One such technology is whereby one high quality cow could be made to produce up to 32 embryos ...

  3. Debating elective single embryo transfer after in vitro fertilization: a ...

    African Journals Online (AJOL)

    However, despite clinical recommendations and policy statements, patients in clinical practice frequently do request for the transfer of multiple embryos in order to have twins. Such requests conflict with policy guidelines and create an ethical dilemma for physicians: Should the physician do as the couple requests, and there ...

  4. Neonatal health including congenital malformation risk of 1072 children born after vitrified embryo transfer.

    Science.gov (United States)

    Belva, F; Bonduelle, M; Roelants, M; Verheyen, G; Van Landuyt, L

    2016-07-01

    transfer. Data were analysed by multiple linear and logistic regression, adjusted for treatment variables and maternal characteristics. Mothers to infants in the vitrified group were on average slightly older and more often suffering from pregnancy-related hypertensive disorders than mothers to infants in the fresh transfer group. Singletons born after vitrification showed a higher birthweight standard deviation score (SDS) (-0.4 versus -0.7; 95% confidence interval (CI): 0.0-0.3, P = 0.001) and a lower small-for-gestational age rate (AOR: 0.55; 95% CI: 0.34-0.90) in comparison with peers born after fresh embryo transfer. Preterm birth rate and perinatal death rate were comparable between the two groups (AOR: 0.91; 95% CI: 0.57-1.43 and AOR: 0.97; 95% CI: 0.40-2.36). In twins, neonatal outcomes including birthweight SDS, small-for-gestational age and prematurity rates were comparable in the vitrified and the fresh groups, when adjusted for confounders. Furthermore, the rate of major congenital malformations in live borns was comparable between the vitrified group and the fresh group, both in singletons (2.6 versus 2.8%; AOR: 0.91; 95% CI: 0.47-1.78) and in twins (2.4 versus 2.7%; AOR: 0.51; 95% CI: 0.05-5.72). Also, the total malformation rate in the vitrified group (3.4%; 95% CI: 2.4-4.8) did not differ from the rate in the fresh embryo group (3.9%; 95% CI: 3.1-5.0). The embryonic stage at vitrification or fresh transfer (cleavage-stage embryo or blastocyst) did not influence the birth characteristics or malformation rate. The main limitation of this study is the rather small twin group. Therefore, the outcome results for twins should be interpreted cautiously. This study provides evidence that transfer of vitrified Day 3 and Day 5 embryos does not adversely affect the neonatal health of the offspring in comparison with transfer of fresh embryos. Furthermore, neonatal outcomes were not different after transfer of vitrified blastocysts compared with transfer of vitrified

  5. The Influence of Interspecies Somatic Cell Nuclear Transfer on Epigenetic Enzymes Transcription in Early Embryos

    DEFF Research Database (Denmark)

    Morovic, Martin; Murin, Matej; Strejcek, Frantisek

    2016-01-01

    One of the main reason for the incorrect development of embryos derived from somatic cell nuclear transfer is caused by insufficient demethylation of injected somatic chromatin to a state comparable with an early embryonic nucleus. It is already known that the epigenetic enzymes transcription....... In spite of the detection of ooplasmic DNA methyltransferases, the somatic genes for DNMT1 and DNMT3a enzymes were not expressed and the development of intergeneric embryos stopped at the 4-cell stage. Our results indicate that the epigenetic reprogramming during early mammalian development is strongly...

  6. Efficiency of porcine somatic cell nuclear transfer – a retrospective study of factors related to embryo recipient and embryos transferred

    Directory of Open Access Journals (Sweden)

    Yongye Huang

    2013-10-01

    The successful generation of pigs via somatic cell nuclear transfer depends on reducing risk factors in several aspects. To provide an overview of some influencing factors related to embryo transfer, the follow-up data related to cloned pig production collected in our laboratory was examined. (i Spring showed a higher full-term pregnancy rate compared with winter (33.6% vs 18.6%, P = 0.006. Furthermore, a regression equation can be drawn between full-term pregnancy numbers and pregnancy numbers in different months (y = 0.692x−3.326. (ii There were no significant differences detected in the number of transferred embryos between surrogate sows exhibiting full-term development compared to those that did not. (iii Non-ovulating surrogate sows presented a higher percentage of full-term pregnancies compared with ovulating sows (32.0% vs 17.5%, P = 0.004; respectively. (iv Abortion was most likely to take place between Day 27 to Day 34. (v Based on Life Table Survival Analysis, delivery in normally fertilized and surrogate sows is expected to be completed before Day 117 or Day 125, respectively. Additionally, the length of pregnancy in surrogate sows was negatively correlated with the average litter size, which was not found for normally fertilized sows. In conclusion, performing embryo transfer in appropriate seasons, improving the quality of embryos transferred, optimizing the timing of embryo transfer, limiting the occurrence of abortion, combined with ameliorating the management of delivery, is expected to result in the harvest of a great number of surviving cloned piglets.

  7. Piglets produced by transfer of vitrified porcine embryos after stepwise dilution of cryoprotectants.

    Science.gov (United States)

    Kobayashi, S; Takei, M; Kano, M; Tomita, M; Leibo, S P

    1998-02-01

    A total of 498 porcine embryos at various stages of development collected from superovulated gilts was used to investigate cryopreservation. First, blastocysts (BL), expanded blastocysts (ExB), and hatched blastocysts (HB) were used to determine the effect of exposure to concentrated solutions of ethylene glycol as cryoprotective additives (CPAs) on embryo survival. Then, survival of other embryos after vitrification by rapid cooling was determined. Based on their development after 48 h in culture, embryos were not injured by being exposed to 2.0 M ethylene glycol (EG) for 15 min or to 2.0 M EG for 5 min and then to a solution of 8.0 M EG in 7% polyvinylpyrrolidone (PVP) for 1 min. The CPAs were removed from the embryos by diluting them with 1.7 M galactose. To vitrify the embryos, they were exposed to 2.0 M EG for 5 min and then were pipetted directly into short columns of 8.0 M EG-PVP contained within (1.25-ml plastic straws and separated from long columns of 1.7 M galactose by an air bubble. The straws were plunged directly into LN2. After the straws were warmed rapidly in a 25 degrees C water bath, the embryos were immediately mixed with galactose within the straws by shaking them vigorously to mix the contents. In sequential experiments, three methods were used to dilute the CPA solutions. Method 1: Embryos in the EG-PVP-galactose mixture were expelled from the straws and rinsed and cultured in modified CZB medium (mCZB). Method II: Embryos in the mixture were placed briefly into 1.5 M EG and then rinsed and cultured in mCZB. Method III: Embryos in the mixture were rinsed in 1.0 M EG and then in 0.5 M EG and finally rinsed with mCZB and cultured. After 48 h in culture, the respective percentages of survival of embryos vitrified as BL, ExB, or HB were: Method I, 21, 32, and 13%; Method II, 9, 40, and 24%; Method III, 35, 85, and 71%. Of 20 additional ExB vitrified embryos diluted by Method III and transferred into a recipient, four developed into live piglets

  8. Transfering vitamin C from fish to embryos

    Science.gov (United States)

    Beneficial effects of ascorbic acid supplementation to broodstock of a select aquaculture species is well documented. At the present levels of feeding, dietary means of vitamin C does not meet the requirements for maturation, reproduction and needs of early life stages of larvae. In addition, thi...

  9. Effect of chilling on the development of in vitro produced bovine embryos at various cleavage stages

    OpenAIRE

    Balasubramanian, S.; Rho, Gyu-Jin

    2006-01-01

    Purpose: Bovine embryos and zygotes are known to be sensitive to “temperature shock” when cooled to temperatures near 0°C. The effect of chilling on in vitro derived embryos at various cleavage stages was investigated.

  10. The effect of flurbiprofen on the development of anencephaly in early stage chicken embryos.

    Science.gov (United States)

    Özeren, Ersin; Er, Uygur; Güvenç, Yahya; Demirci, Adnan; Arıkök, Ata Türker; Şenveli, Engin; Ergün, Rüçhan Behzat

    2015-04-01

    The study investigated the effect of flurbiprofen on the development of anencephaly in early stage chicken embryos. We looked at four groups with a total of 36 embryos. There was a control group, a normal saline group, a normal-dose group and a high-dose group with ten, ten, eight and eight eggs with embryo respectively. Two embryos in the control group, studied with light microscopy at 48 h, were consistent with 28-29 hours' incubation in the Hamburger-Hamilton System. They had open neural tubes. The other embryos in this group were considered normal. One embryo in the normal saline group was on the occlusion stage at 48 h. One embryo showed an open neural tube. They were compatible with 28-29 hours' incubation in the Hamburger-Hamilton system. The remaining eight embryos showed normal development. In the normal dose group, one embryo showed underdevelopment of the embryonic disc and the embryo was dead. In four embryos, the neural tubes were open. One cranial malformation was found that was complicated with anencephaly in one embryo. In two embryos the neural tubes were closed, as they showed normal development, and they reached their expected stages according to the Hamburger-Hamilton classification. There was no malformation or growth retardation. Four experimental embryos were anencephalic in the high dose group, and three embryos had open neural tubes. One embryo exhibited both anencephaly and a neural tube closure defect. None of the embryos in this group showed normal development. Even the usual therapeutic doses of flurbiprofen increased the risk of neural tube defect. Flurbiprofen was found to significantly increase the risk of anencephaly. The provision of improved technical materials and studies with larger sample sizes will reveal the stage of morphological disruption during the development of embryos.

  11. Prediction of in-vitro developmental competence of early cleavage-stage mouse embryos with compact time-lapse equipment.

    Science.gov (United States)

    Pribenszky, Csaba; Losonczi, Eszter; Molnár, Miklós; Lang, Zsolt; Mátyás, Szabolcs; Rajczy, Klára; Molnár, Katalin; Kovács, Péter; Nagy, Péter; Conceicao, Jason; Vajta, Gábor

    2010-03-01

    Single blastocyst transfer is regarded as an efficient way to achieve high pregnancy rates and to avoid multiple pregnancies. Risk of cancellation of transfer due to a lack of available embryos may be reduced by early prediction of blastocyst development. Time-lapse investigation of mouse embryos shows that the time of the first and second cleavage (to the 2- and 3-cell stages, respectively) has a strong predictive value for further development in vitro, while cleavage from the 3-cell to the 4-cell stage has no predictive value. In humans, embryo fragmentation during preimplantation development has been associated with lower pregnancy rates and a higher incidence of developmental abnormalities. Analysis of time-lapse records shows that most fragmentation is reversible in the mouse and is resorbed in an average of 9 h. Daily or bi-daily microscopic checks of embryo development, applied routinely in human IVF laboratories, would fail to detect 36 or 72% of these fragmentations, respectively. Fragmentation occurring in a defined time frame has a strong predictive value for in-vitro embryo development. The practical compact system used in the present trial, based on the 'one camera per patient' principle, has eliminated the usual disadvantages of time-lapse investigations and is applicable for the routine follow-up of in-vitro embryo development. Copyright 2009 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  12. Identification of abnormal gene expression in bovine transgenic somatic cell nuclear transfer embryos

    OpenAIRE

    Cho, Jongki; Kang, Sungkeun; Lee, Byeong Chun

    2014-01-01

    This study was conducted to investigate the expression of three genes related to early embryonic development in bovine transgenic cloned embryos. To accomplish this, development of bovine transgenic somatic cell nuclear transfer (SCNT) embryos was compared with non-transgenic embryos. Next, mRNA transcription of three specific genes (DNMT1, Hsp 70.1, and Mash2) related to early embryo development in transgenic SCNT embryos was compared between transgenic and non-transgenic SCNTs, parthenogene...

  13. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF

    NARCIS (Netherlands)

    Vergouw, C.G.; Kostelijk, E.H.; Doejaaren, E.; Hompes, P.G.A.; Lambalk, C.B.; Schats, R.

    2012-01-01

    STUDY QUESTION Does the type of medium used to culture fresh and frozenthawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? SUMMARY ANSWER A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean

  14. Pregnancy outcomes in a university hospital after legal requirement for single-embryo transfer.

    Science.gov (United States)

    Ercan, Cihangir Mutlu; Kerimoglu, Ozlem Secilmis; Sakinci, Mehmet; Korkmaz, Cem; Duru, Namik Kemal; Ergun, Ali

    2014-04-01

    To present our in vitro fertilization outcomes after implementation of the Turkish government policy limiting the number of transferred embryos to one, in women under the age of 35, and two, in women over age 35; and to demonstrate the feasibility of this policy. We retrospectively reviewed the records of 423 patients who underwent ovarian stimulation, oocyte pick-up, intracytoplasmic sperm injection and embryo transfer due to primary or secondary infertility over a 22-month time interval. Clinical pregnancy was defined as a fetal heartbeat in the seventh gestational week. Embryo transfers were carried out on day 3 or 5. Descriptive statistics are given as the number of subjects and percentages. A total of 353 embryo transfers were performed: 261 (73.9%) were single-embryo transfer (244 (69.1%) elective and 17 (4.8%) non-elective) and 92 (26.1%) were double-embryo transfer. Of the 244 elective single-embryo transfers, 6.6% (n=16) were performed using frozen-thawed embryos. The average patient age was 29.6±4.5 years in the single-embryo group and 36.5±2.4 years in the double-embryo group. The cumulative pregnancy rates per oocyte pick-up were similar in both groups: 41% (n=107) in the single-embryo group and 43.4% (n=40) in the double-embryo group (p=0.678). The cumulative live birth rate of the single-embryo group (32.1%) was not statistically different from the double-embryo group (35.8%) (p=0.518). The twin pregnancy rate after single-embro transfer was significantly lower than with double-embryo transfer (2.8% (n=3) vs. 32.5% (n=13); ppregnancies occurred in women 35 years or older who underwent double-embryo transfer. No significant difference in the spontaneous abortion rates was recorded between the single- and double-embryo transfer groups (16 (6.1%) vs. 6 (6.5%); p=0.894). Single-embryo transfer results in a pregnancy rate comparable to double-embryo transfer, with a significantly reduced multiple pregnancy rate. Double-embryo transfer in patients over

  15. Influence of embryo handling and transfer method on pig cloning efficiency.

    Science.gov (United States)

    Shi, Junsong; Zhou, Rong; Luo, Lvhua; Mai, Ranbiao; Zeng, Haiyu; He, Xiaoyan; Liu, Dewu; Zeng, Fang; Cai, Gengyuan; Ji, Hongmei; Tang, Fei; Wang, Qinglai; Wu, Zhenfang; Li, Zicong

    2015-03-01

    The somatic cell nuclear transfer (SCNT) technique could be used to produce genetically superior or genetically engineered cloned pigs that have wide application in agriculture and bioscience research. However, the efficiency of porcine SCNT currently is very low. Embryo transfer (ET) is a key step for the success of SCNT. In this study, the effects of several ET-related factors, including cloned embryo culture time, recipient's ovulation status, co-transferred helper embryos and ET position, on the success rate of pig cloning were investigated. The results indicated that transfer of cloned embryos cultured for a longer time (22-24h vs. 4-6h) into pre-ovulatory sows decreased recipient's pregnancy rate and farrowing rate, and use of pre-ovulatory and post-ovulatory sows as recipients for SCNT embryos cultured for 22-24h resulted in a similar porcine SCNT efficiency. Use of insemination-produced in vivo fertilized, parthenogenetically activated and in vitro fertilized embryos as helper embryos to establish and/or maintain pregnancy of SCNT embryos recipients could not improve the success rate of porcine SCNT. Transfer of cloned embryos into double oviducts of surrogates significantly increased pregnancy rate as well as farrowing rate of recipients, and the developmental rate of transferred cloned embryos, as compared to unilateral oviduct transfer. This study provided useful information for optimization of the embryo handling and transfer protocol, which will help to improve the ability to generate cloned pigs. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Development of interspecies nuclear transfer embryos reconstructed with argali (Ovis ammon) somatic cells and sheep ooplasm.

    Science.gov (United States)

    Pan, Xiaoyan; Zhang, Yanli; Guo, Zhiqin; Wang, Feng

    2014-02-01

    Interspecies nuclear transfer has already achieved success in several species, which shows great potential in recovery and conservation of endangered animals. The study was conducted to establish an efficient system for in vitro argali (Ovis ammon)-sheep embryo reconstruction via interspecies somatic cell nuclear transfer (iSCNT). The competence of domestic sheep cytoplasts to reprogram the adult argali fibroblast nuclei was evaluated, and the effects of enucleation methods and donor cell passage and cell state on the in vitro development of argali-sheep cloned embryos were also examined. Sheep oocytes could support argali and sheep fibroblast cell nuclei transfer and develop to blastocysts in vitro. Oocytes matured for 21–23 h and enucleated by chemically assisted enucleation (CAE) had a higher enucleation rate than blind enucleation (BE), but the development rate of iSCNTembryos was the same (P>0.05). Moreover, passage numbers of fibroblast cells embryos. Thus sheep cytoplasm successfully supports argali nucleus development to blastocyst stage after optimising the nuclear transfer procedure, which indicates that iSCNT can be used to conserve endangered argali in the near future.

  17. Histone deacetylase inhibitor significantly improved the cloning efficiency of porcine somatic cell nuclear transfer embryos.

    Science.gov (United States)

    Huang, Yongye; Tang, Xiaochun; Xie, Wanhua; Zhou, Yan; Li, Dong; Yao, Chaogang; Zhou, Yang; Zhu, Jianguo; Lai, Liangxue; Ouyang, Hongsheng; Pang, Daxin

    2011-12-01

    Valproic acid (VPA), a histone deacetylase inbibitor, has been shown to generate inducible pluripotent stem (iPS) cells from mouse and human fibroblasts with a significant higher efficiency. Because successful cloning by somatic cell nuclear transfer (SCNT) undergoes a full reprogramming process in which the epigenetic state of a differentiated donor nuclear is converted into an embryonic totipotent state, we speculated that VPA would be useful in promoting cloning efficiency. Therefore, in the present study, we examined whether VPA can promote the developmental competence of SCNT embryos by improving the reprogramming state of donor nucleus. Here we report that 1 mM VPA for 14 to 16 h following activation significantly increased the rate of blastocyst formation of porcine SCNT embryos constructed from Landrace fetal fibroblast cells compared to the control (31.8 vs. 11.4%). However, we found that the acetylation level of Histone H3 lysine 14 and Histone H4 lysine 5 and expression level of Oct4, Sox2, and Klf4 was not significantly changed between VPA-treated and -untreated groups at the blastocyst stage. The SCNT embryos were transferred to 38 surrogates, and the cloning efficiency in the treated group was significantly improved compared with the control group. Taken together, we have demonstrated that VPA can improve both in vitro and in vivo development competence of porcine SCNT embryos.

  18. The effects of MRI on mouse embryos during fetal stage

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Takashi; Sakazaki, Takahiko; Itokawa, Yuka [Suzuka University of Medical Science, Koriyama (Japan)] (and others)

    2006-06-15

    The effects of Magnetic Resonance Imaging (MRI) on mouse embryos at the early stage of organogenesis were investigated. Pregnant ICR mice were exposed on day 8 of gestation to MRI at 0.5 T for 0.5 hour to 3 hours. The mortality rates of embryos or fetuses, the incidence of external malformations, fetal body weight and sex ratio were observed at day 18 of gestation. A significant increase in embryonic mortality was observed after exposure to either 0.5 T MRI for 0.5 hour or 2 hours. However, the exposure to MRI for 1 hour or 3 hours did not induce any significant increase in embryonic mortality when compared with control. External malformations such as exencephaly, cleft palate and anomalies of tail were observed in all experimental groups exposed to each MRI. A statistically significant increase of external malformations was observed in all groups treated with 0.5 T MRI for 0.5 hour and 3 hours. The incidence of external malformations in the mice group exposed to 0.5 T MRI for 0.5-hour was found to be higher than those of mice group exposed to 0.5 T MRI for 2 hours. The effects of MRI on the external malformations might not to be dose-dependent. There was no statistically significant difference in fetal body weight and sex ratio among each MRI exposure groups.

  19. Comparison of pregnancy rate between fresh embryo transfers and frozen-thawed embryo transfers following ICSI treatment

    Directory of Open Access Journals (Sweden)

    Zahra Basirat

    2016-01-01

    Full Text Available Background: The use of assisted reproductive technology (ART is increasing in the world. The rate, efficacy and safety of ART are very different among countries. There is an increase in the use of intra cytoplasmic sperm injection (ICSI, single fresh embryo transfer (ET and frozen-thawed embryo transfer (FET. Objective: The objective of this study was to compare pregnancy rate in fresh ET and FET. Materials and Methods: In this retrospective cross-sectional study 1014 ICSI-ET cycles (426 fresh ET and 588 FET from 753 women undergoing ICSI treatment referred to Fatemezahra Infertility and Reproductive Health Research Center in Babol, Iran from 2008 to 2013 were reviewed. Results: There were no significant differences between biochemical pregnancy rate (23% versus 18.8%, OR 1.301; 95% CI .95-1.774, gestational sac (95.6% versus 100% in FET, OR 0.60; 95% CI 0.54-0.67, and fetal heart activity (87.2% versus 93.6% OR .46; 95% CI .16-1.32 in fresh ET and FET cycles, respectively. P< 0.05 was considered statistically significant for all measures. Conclusion: Although, the result showed no significantly difference between the fresh ET and the FET cycles, however the embryos are able to be stored for subsequent ART. Therefore, we recommend FET cycles as an option alongside the fresh ET.

  20. Influence of beta-carotene on fertility in rabbits when using embryo transfer programs.

    Science.gov (United States)

    Besenfelder, U; Solti, L; Seregi, J; Brem, G

    1993-05-01

    The effect of beta-carotene on reproduction traits in rabbits was studied in 509 (superovulated and normally ovulated) donors and 239 recipients by using embryo/gene transfer performed at 2 different locations. All of the bucks and the half of the females were fed a diet supplemented with 40 mg synthetic beta-carotene (Rovimix((R)))/kg feed. Embryos at the pronucleus stage were collected 19 to 21 hours after induction of ovulation with human chorionic gonadotropin (hCG); they were then microinjected into the male pronucleus and transferred to synchronized recipients. Data were obtained from the time when the donors and recipients were caged, until the pups resulting from the embryo transfers were weaned. Supplemented beta-carotene did not affect most of the 30 traits that were analyzed. However superovulated donors in Project 2 that received the beta-carotene enriched diet had a 14% lighter ovary weight (P<0.05) and less than half of the oocytes were unfertilized (P<0.05). In Project 1 (beta-carotene group) there was a greater number of pups born (36%, P<0.05) and more of these pups were born alive (53%, P<0.05).

  1. Full-term development of gaur-bovine interspecies somatic cell nuclear transfer embryos: effect of trichostatin A treatment.

    Science.gov (United States)

    Srirattana, Kanokwan; Imsoonthornruksa, Sumeth; Laowtammathron, Chuti; Sangmalee, Anawat; Tunwattana, Wanchai; Thongprapai, Thamnoon; Chaimongkol, Chockchai; Ketudat-Cairns, Mariena; Parnpai, Rangsun

    2012-06-01

    Trichostatin A (TSA) has previously been used in somatic cell nuclear transfer (SCNT) to improve the cloning efficiency in several species, which led our team to investigate the effects of TSA on the full-term development of bovine SCNT and gaur-bovine interspecies SCNT (gaur iSCNT; gaur somatic cells as donors and bovine oocytes as recipients) embryos. Treatment with 50 nM TSA for 10 h after fusion had no positive effects on the rates of fusion, cleavage, or the development to eight-cell or morula stages in both bovine SCNT and gaur iSCNT embryos. However, TSA treatment significantly enhanced the blastocyst formation rate in bovine SCNT embryos (44 vs. 32-34% in the TSA-treated and TSA-untreated groups, respectively), but had no effects on gaur iSCNT embryos. The fresh blastocysts derived from bovine SCNT and gaur iSCNT embryos (fresh groups), as well as vitrified bovine SCNT blastocysts (vitrified group), were transferred to bovine recipients. We found that TSA treatment increased the pregnancy rates only in recipients receiving fresh bovine SCNT embryos. In recipients receiving TSA-treated bovine SCNT embryos, three cloned calves from the fresh group and twin cloned calves from the vitrified group were delivered; however, no calf was born from the TSA-untreated bovine SCNT embryos. In contrast, one gaur iSCNT calf was born from a recipient receiving blastocysts from the TSA-untreated group. In summary, TSA improved the preimplantation development and pregnancy rates of bovine SCNT embryos, but did not have any beneficial effect on gaur iSCNT embryos. However, one gaur iSCNT calf reached full-term development.

  2. Effect of embryo age and recipient asynchrony on pregnancy rates in a commercial equine embryo transfer program.

    Science.gov (United States)

    Jacob, J C F; Haag, K T; Santos, G O; Oliveira, J P; Gastal, M O; Gastal, E L

    2012-04-01

    In the present study, 809 uterine flushes and 454 embryo transfers performed in mares over a 4-yr interval were examined to evaluate the effects of: (1) the day of embryo collection on recovery rates; (2) the degree of synchrony between donor and recipient mares on pregnancy rates; (3) the recipient day post ovulation on pregnancy rates; and (4) the age of the embryo at recovery on pregnancy rates at 60 days. Uterine flushes were performed on Days 6, 7, 8, 9, and 10 (Day 0 = ovulation) and embryos were transferred to recipients with degrees of synchrony varying between +1 to -6 (recipient ovulated 1 day before through 6 days after the donor). Recipient mares ranged from 2 to 8 days post ovulation. Embryo recovery rates were similar for flushes performed on Day 7 (61%), Day 8 (66%), Day 9 (59%), and Day 10 (56%), but the embryo recovery rate was lower (P recipient mares on Day 2 (33%) compared with mares on Day 3 (66%), Day 4 (66%), Day 5 (62%), Day 6 (55%), Day 7 (58%), and Day 8 (56%). Pregnancy rate was higher (P recipient mares does not need to be as restricted as previously reported in horses. Acceptable pregnancy rates (e.g., 70%, 99/142) were obtained even when recipient mares ovulated 4 to 5 days after the donors; (3) similar pregnancy rates were obtained when recipient mares received embryos within a large range of days post ovulation (Days 3 to 8); and (4) Day 7 embryos produced higher pregnancy rates when compared with Days 8 and 9 embryos. In clinical terms, the application of these new findings will be beneficial to large equine embryo transfer operations in producing more pregnancies per season. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Yongye; Tang, Xiaochun; Xie, Wanhua; Zhou, Yan; Li, Dong; Zhou, Yang; Zhu, Jianguo; Yuan, Ting; Lai, Liangxue [Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun 130062 (China); Pang, Daxin, E-mail: pdx@jlu.edu.cn [Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun 130062 (China); Ouyang, Hongsheng, E-mail: ouyh@jlu.edu.cn [Jilin Province Key Laboratory of Animal Embryo Engineering, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi An DaLu, Changchun 130062 (China)

    2011-07-29

    Highlights: {yields} Report for the first time that vitamin C has a beneficial effect on the development of porcine SCNT embryos. {yields} The level of acH4K5 and Oct4 expression at blastocyst-stage was up-regulated after treatment. {yields} A higher rate of gestation and increased number of piglets born were harvested in the treated group. -- Abstract: The reprogramming of differentiated cells into a totipotent embryonic state through somatic cell nuclear transfer (SCNT) is still an inefficient process. Previous studies revealed that the generation of induced pluripotent stem (iPS) cells from mouse and human fibroblasts could be significantly enhanced with vitamin C treatment. Here, we investigated the effects of vitamin C, to our knowledge for the first time, on the in vitro and in vivo development of porcine SCNT embryos. The rate of blastocyst development in SCNT embryos treated with 50 {mu}g/mL vitamin C 15 h after activation (36.0%) was significantly higher than that of untreated SCNT embryos (11.5%). The enhanced in vitro development rate of vitamin C-treated embryos was associated with an increased acetylation level of histone H4 lysine 5 and higher Oct4, Sox2 and Klf4 expression levels in blastocysts, as determined by real-time PCR. In addition, treatment with vitamin C resulted in an increased pregnancy rate in pigs. These findings suggest that treatment with vitamin C is beneficial for enhancement of the in vitro and in vivo development of porcine SCNT embryos.

  4. Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos

    International Nuclear Information System (INIS)

    Huang, Yongye; Tang, Xiaochun; Xie, Wanhua; Zhou, Yan; Li, Dong; Zhou, Yang; Zhu, Jianguo; Yuan, Ting; Lai, Liangxue; Pang, Daxin; Ouyang, Hongsheng

    2011-01-01

    Highlights: → Report for the first time that vitamin C has a beneficial effect on the development of porcine SCNT embryos. → The level of acH4K5 and Oct4 expression at blastocyst-stage was up-regulated after treatment. → A higher rate of gestation and increased number of piglets born were harvested in the treated group. -- Abstract: The reprogramming of differentiated cells into a totipotent embryonic state through somatic cell nuclear transfer (SCNT) is still an inefficient process. Previous studies revealed that the generation of induced pluripotent stem (iPS) cells from mouse and human fibroblasts could be significantly enhanced with vitamin C treatment. Here, we investigated the effects of vitamin C, to our knowledge for the first time, on the in vitro and in vivo development of porcine SCNT embryos. The rate of blastocyst development in SCNT embryos treated with 50 μg/mL vitamin C 15 h after activation (36.0%) was significantly higher than that of untreated SCNT embryos (11.5%). The enhanced in vitro development rate of vitamin C-treated embryos was associated with an increased acetylation level of histone H4 lysine 5 and higher Oct4, Sox2 and Klf4 expression levels in blastocysts, as determined by real-time PCR. In addition, treatment with vitamin C resulted in an increased pregnancy rate in pigs. These findings suggest that treatment with vitamin C is beneficial for enhancement of the in vitro and in vivo development of porcine SCNT embryos.

  5. Methods employed to overcome difficult embryo transfer during assisted reproduction treatment.

    Science.gov (United States)

    Akhtar, M A; Netherton, R; Majumder, K; Edi-Osagie, E; Sajjad, Y

    2015-08-01

    Multiple steps, at both scientific and clinical level, are involved in assisted reproduction technology (ART). Optimization of each of these steps contributes to maximize the success of ART. Embryo transfer is one of the vital steps in ART. An easy embryo transfer increases the success of ART. Adequate training is required to undertake embryo transfer, but anatomical difficulties in completing this procedure alter the success of treatment. Difficult embryo transfer is challenging for clinicians but has an overwhelming negative impact on patients. Difficult embryo transfer may cause cervical or endometrial trauma with uterine contractions which can lead to unsuccessful implantation and poor outcome after ART. Literature review of published material looking at all interventions that were employed to overcome difficult embryo transfers during ART. We identified 5 methods (17 studies) were employed to overcome difficult embryo transfer which are: Hysteroscopic methods in 4 studies (2 case reports, 2 case series). Malecot catheters after hysteroscopic evaluation were used in 2 studies (1 case report, 1 case series). Dilators including hygroscopic dilators were used in 2 studies (1 case report, 1 case series) and mechanical dilators were used in 3 studies (2 case series, 1 RCT-patients 367). Intrafollopian transfer was reported in 1 case report. Transmyometrial embryo transfers were reported in 5 studies (2 case reports, 2 case series, 1 RCT). There were only two randomized controlled studies (RCTs) identified in the review. All other were either case series or case reports. We found that there was no uniform classification or grading of difficulty of embryo transfer in the literature. A grading system has been suggested in this review. We recommend that there should be a consensus guideline formulated for interventions to overcome difficult embryo transfer. A large multicenter randomized controlled study is required to compare different methods for women with difficult

  6. Production of transgenic canine embryos using interspecies somatic cell nuclear transfer.

    Science.gov (United States)

    Hong, So Gun; Oh, Hyun Ju; Park, Jung Eun; Kim, Min Jung; Kim, Geon A; Koo, Ok Jae; Jang, Goo; Lee, Byeong Chun

    2012-02-01

    Somatic cell nuclear transfer (SCNT) has emerged as an important tool for producing transgenic animals and deriving transgenic embryonic stem cells. The process of SCNT involves fusion of in vitro matured oocytes with somatic cells to make embryos that are transgenic when the nuclear donor somatic cells carry 'foreign' DNA and are clones when all the donor cells are genetically identical. However, in canines, it is difficult to obtain enough mature oocytes for successful SCNT due to the very low efficiency of in vitro oocyte maturation in this species that hinders canine transgenic cloning. One solution is to use oocytes from a different species or even a different genus, such as bovine oocytes, that can be matured easily in vitro. Accordingly, the aim of this study was: (1) to establish a canine fetal fibroblast line transfected with the green fluorescent protein (GFP) gene; and (2) to investigate in vitro embryonic development of canine cloned embryos derived from transgenic and non-transgenic cell lines using bovine in vitro matured oocytes. Canine fetal fibroblasts were transfected with constructs containing the GFP and puromycin resistance genes using FuGENE 6®. Viability levels of these cells were determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. Interspecies SCNT (iSCNT) embryos from normal or transfected cells were produced and cultured in vitro. The MTT measurement of GFP-transfected fetal fibroblasts (mean OD = 0.25) was not significantly different from non-transfected fetal fibroblasts (mean OD = 0.35). There was no difference between transgenic iSCNT versus non-transgenic iSCNT embryos in terms of fusion rates (73.1% and 75.7%, respectively), cleavage rates (69.7% vs. 73.8%) and development to the 8-16-cell stage (40.1% vs. 42.7%). Embryos derived from the transfected cells completely expressed GFP at the 2-cell, 4-cell, and 8-16-cell stages without mosaicism. In summary, our results demonstrated that

  7. EMBRYOSPLITTING - A WAY TO INCREASE THE EFFICIENCY OF EMBRYO - TRANSFER IN SHEEP

    OpenAIRE

    Vintila, I.; Benscik, I.; Pacala, N.; Corin, N.; Babusik, I.; Kulickova, L.

    1996-01-01

    In has been compared the gestation results and the number of lambs obtained from 40 unsplitted integral embryos collected from 31 superovulated sheep, to the same parameters in the situation when 40 demi -embryos were used, in order to estimate the embryosplitting advantage to increase the efficiency or embryo- transfer in sheep. The gestation percentage obtained by the recipient sheep in the case of integral embryos, was 57,5% and in case of demiembryos was 47,5%, only. Also, the proportion ...

  8. Optimal developmental stage for vitrification of parthenogenetically activated porcine embryos

    DEFF Research Database (Denmark)

    Li, Rong; Li, Juan; Kragh, Peter

    2012-01-01

    -lapse monitored for 24 h or analyzed by diffential staining. After warming, the embryos had to be cultured for at least 8 h before their survival rates were stabilized. Both the survival rate and 8 h and the hatching rate at 24 h of Day 4 embryos were significantly higher than those vitrified on Day5 or Day 6 (P...

  9. Monozygotic Triplets and a Singleton After ICSI and Day 3 Transfer of Two Embryos

    Directory of Open Access Journals (Sweden)

    Singpetch Suksompong

    2017-07-01

    Full Text Available Monozygotic triplets are a rare condition to occur naturally or even in assisted reproductive technologies. In this report, we present a case of monozygotic triplets with a singleton pregnancy following embryo transfer of 2 embryos. To our knowledge, this is the first published case of one singleton and monozygotic triplets after ICSI and transfer of two day 3 embyos in Thailand.

  10. Non-surgical transfer of 4',6'-diamidino-2-phenylindole-stained equine embryos

    Directory of Open Access Journals (Sweden)

    M. HUHTINEN

    2008-12-01

    Full Text Available 4',6'-diamidino-2-phenylindole (DAPI is a fluorescent dye that binds only to the DNA of dead blastomeres of an embryo. The use of DAPI for assessing viabilities of equine Day 6 to 7 embryos was studied. The pregnancy and normal development of the embryo until 20 days after transfer (day of pregnancy termination was used as an indicator of embryonic viability. Eleven embryos were stained with DAPI at room temperature for 15 min. They were then exposed to UV light to visualize staining, and cultured for 2 h before non-surgical transfer to recipient mares. Eight control embryos were cultured for 2 h before transfer to recipient mares. The recipient mares were scanned for pregnancies every other day starting 6 days after transfer. Twenty days after the transfer the pregnant recipient mares received luprostiol to induce abortion of the foetus. The pregnancy rate 6 days after transfer was 82% in the treatment group, and 75% in the control group (P>0.05. One DAPI-stained embryo resorbed and was no longer visible 12 days after transfer. However, this particular recipient mare had a uterine inflammation as shown by examination in the oestrus following embryo transfer. Although the number of embryos is insufficient to demonstrate lack of treatment effect, the high pregnancy rates following DAPI staining should encourage further studies on this subject.;

  11. Successful pregnancy rates achieved with day 4 embryo transfers.

    Science.gov (United States)

    Skorupski, Josh C; Stein, Daniel E; Acholonu, Uchenna; Field, Heather; Keltz, Martin

    2007-04-01

    To assess the success of day 4 embryo transfers (ETs) following IVF at one institution. Retrospective analysis. A university hospital IVF program. Two hundred nondonor, fresh IVF cycles. None. Outcomes of IVF. Outcome assessments after day 4 ETs included rates of implantation, clinical pregnancy, and singleton and multiple live births. The overall live-birth rate was 54.4%. Implantation rates were highest in younger age groups, and similar in patients 35-40 years of age. Pregnancy and live-birth rates were similar across all age groups up to age 40 years. Multiple gestations were highest in women < or =40 years of age. Acceptable pregnancy rates can be achieved with day 4 ETs.

  12. IVF and embryo transfer: historical origin and development.

    Science.gov (United States)

    Biggers, John D

    2012-08-01

    IVF and embryo transfer for the treatment of human infertility has now resulted in the birth of over 4 million babies. The technique did not arise as a quantum event but was built on the efforts of many earlier workers in the fields of reproductive endocrinology and development. One should remember the famous saying of Isaac Newton: 'If I have seen further than most, it is because I have stood on the shoulder's of giants'. Ethical and moral issues have always arisen when investigators study early mammalian development, particularly human development. This paper documents these earlier studies and also draws attention to the ethical and moral arguments that inevitably arose. Copyright © 2012 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  13. Does the number of oocytes retrieved influence pregnancy after fresh embryo transfer?

    Directory of Open Access Journals (Sweden)

    Qianfang Cai

    Full Text Available BACKGROUND: The nature of the association between the number of oocytes retrieved and in vitro fertilization (IVF outcomes after fresh embryo transfer remains unclear because of conflicting results reported in the studies on this subject. In addition, the influence of the quality of the embryos transferred is usually neglected. The objective of this study is to assess the relationships of the number of oocytes retrieved, the number and quality of embryos transferred, and the prospects of pregnancy after fresh embryo transfer. METHODS: The data on 3131 infertile women undergoing their first IVF treatment cycle between January 2009 and December 2010 were collected retrospectively. Restricted cubic splines and stratified analyses were used to explore the relationships between the number of oocytes retrieved, the number and quality of embryos transferred, and the IVF outcomes. RESULTS: When stratified by the number and quality of transferred embryos, no significant differences in the chances for clinical pregnancy and live birth were found in three groups of oocytes yielded (≤6, 7-14, or ≥15. The relationship between the number of oocytes retrieved and pregnancy is nearly a reflection of the pattern of the relationship between the number of oocytes retrieved and the probability of having two good-quality embryos transferred. The patients with the "optimal" number of oocytes were not only younger but also had the highest probability of having two good-quality embryos replaced. CONCLUSIONS: Similarly aged patients have similar pregnancy prospects after fresh embryo transfer when the same number and quality of embryos are replaced, irrespective of their number of oocytes. Selecting the desired number of good-quality embryos for transfer is the key to IVF success. Thus, aiming at retrieving an optimal number of oocytes to maximize IVF outcomes in a fresh cycle could place undue stress on the patients and may not be the best medical decision.

  14. Role of ooplasm in nuclear and nucleolar remodeling of intergeneric somatic cell nuclear transfer embryos during the first cell cycle

    DEFF Research Database (Denmark)

    Østrup, Olga; Strejcek, Frantisek; Petrovicova, Ida

    2011-01-01

    Initially, development of the zygote is under control of the oocyte ooplasm. However, it is presently unknown if and to what extent is the ooplasm able to interact with a transferred somatic cell from another species in the context of interspecies somatic cell nuclear transfer (SCNT). Here, one-cell...... stage embryos were processed at different points in time post activation (2 hpa, 4 hpa, 8 hpa, and 12 hpa) for detailed nuclear and nucleolar analysis by TEM, and immunofluorescence for visualization of nucleolar proteins related to transcription (UBF) and processing (fibrillarin). Bovine and porcine...... intergeneric SCNT embryos were compared to their parthenogenetic counterparts to assess the effects of the introduced somatic cell. Despite the absence of morphological remodeling (premature chromatin condensation, nuclear envelope breakdown), reconstructed embryos showed nuclear and nucleolar precursor body...

  15. In situ DNA transfer to chicken embryos by biolistics

    Directory of Open Access Journals (Sweden)

    Luciana A. Ribeiro

    1999-12-01

    Full Text Available Fertilized chicken eggs were bombarded with a biolistic device. Transient expression of the lacZ gene under the control of a human cytomegalovirus (CMV promoter was assessed after in situ gene transfer using this approach. The influence of different pressures, vacuum levels and particles was tested. Survival rate improved as particle velocity decreased, but resulted in lower levels of expression. The best survival and expression were obtained with gold particles, a helium gas pressure of 600 psi and a vacuum of 600 mmHg. Under these conditions, all bombarded embryos showed b-galactosidase activity, indicating that this was an effective method for transformation of chicken embryos.Ovos fertilizados de galinha foram bombardeados através da técnica de biobalística. A expressão transiente do gene lacZ, sob o controle do promotor humano citomegalovírus, foi verificada após a transferência in situ. Diferentes níveis de pressão de gás hélio, vácuo e tipos de partículas foram testados. A taxa de sobrevivência aumentou à medida que a velocidade das partículas diminuíram, entretanto, o nível de expressão foi menor. Os melhores resultados, combinando taxa de sobrevivência e expressão, foram obtidos com partículas de ouro, 600 libras por polegada ao quadrado de hélio e 600 mmHg de vácuo. Nestas condições, todos os embriões bombardeados apresentaram atividade da b-galactosidase, indicando que esta técnica é eficiente para a transformação de embriões de galinhas.

  16. Improvement of porcine cloning efficiency by trichostain A through early-stage induction of embryo apoptosis.

    Science.gov (United States)

    Ji, Qianqian; Zhu, Kongju; Liu, Zhiguo; Song, Zhenwei; Huang, Yuankai; Zhao, Haijing; Chen, Yaosheng; He, Zuyong; Mo, Delin; Cong, Peiqing

    2013-03-15

    Trichostain A (TSA), an inhibitor of histone deacetylases, improved developmental competence of SCNT embryos in many species, apparently by improved epigenetic reprogramming. The objective of the present study was to determine the effects of TSA-induced apoptosis in cloned porcine embryos. At various developmental stages, a comet assay and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling staining were used to detect apoptosis, and real-time polymerase chain reaction was used to assess expression of genes related to apoptosis and pluripotency. In this study, TSA significantly induced apoptosis (in a dose-dependent manner) at the one-, two-, and four-cell stages. However, in blastocyst stage embryos, TSA decreased the apoptotic index (P < 0.05). Expression levels of Caspase 3 were higher in TSA-treated versus control embryos at the two-cell stage (not statistically significant). The expression ratio of antiapoptotic Bcl-xl gene to proapoptotic Bax gene, an indicator of antiapoptotic potential, was higher in TSA-treated groups at the one-, two-, and four-cell and blastocyst stages. Furthermore, expression levels of pluripotency-related genes, namely, Oct4 and Nanog, were elevated at the morula stage (P < 0.05) in TSA treatment groups. We concluded that inducing apoptosis might be a mechanism by which TSA promotes development of reconstructed embryos. At the initial stage of apoptosis induction, abnormal cells were removed, thereby enhancing proliferation of healthy cells and improving embryo quality. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. Trends and factors associated with the Day 5 embryo transfer, assisted reproductive technology surveillance, USA, 2001-2009.

    Science.gov (United States)

    Marsh, Courtney A; Farr, Sherry L; Chang, Jeani; Kissin, Dmitry M; Grainger, David A; Posner, Samuel F; Macaluso, Maurizio; Jamieson, Denise J

    2012-08-01

    What characteristics are associated with a Day 5 embryo transfer? The use of the Day 5 embryo transfer has increased over time, with clinicians allowing women with typically 'poorer' prognostic characteristics to undergo a Day 5 embryo transfer. The mean number of embryos per Day 5 transfer decreased from 2001 to 2009, although the prevalence of the Day 5 single embryo transfer remains low and the rate of multiple births remains substantial. Day 5 embryo transfer may reduce the rate of multiple gestation pregnancy. US trends over time in the prevalence of the Day 5 transfer, changes in characteristics of patients receiving Day 5 transfer, and number of embryos transferred are unknown. We used 2001-2009 US National assisted reproductive technology (ART) Surveillance System (NASS) data on 620,295 fresh IVF cycles derived from autologous oocytes with a Day 3 or 5 embryo transfer. Trends in the mean number of embryos transferred from 2001 to 2009 were assessed by the day of transfer. For 349,947 cycles from clinics performing both Days 3 and 5 embryo transfers, multivariable logistic regression was used to determine the characteristics associated with the Day 5 embryo transfer. We also compared the characteristics of the Day 5 embryo cycles in 2001 and 2009. Overall, the proportion of ART cycles using the Day 5 embryo transfer increased from 12% in 2001 to 36% in 2009 (Psingle embryo transfer tripled from 4.5% in 2001 to 14.8% in 2009 (PWomen undergoing multiple ART cycles over time are not linked. We ran multivariable logistic regression to lessen the effects of the confounding factors. Cycle cancelation rates by the day of embryo transfer are unknown. Generalizable to ART clinics included in NASS. This study was funded by the Centres for Disease Control. The authors have no competing interests to declare.

  18. The Early Stages of Heart Development: Insights from Chicken Embryos

    Directory of Open Access Journals (Sweden)

    Johannes G. Wittig

    2016-04-01

    Full Text Available The heart is the first functioning organ in the developing embryo and a detailed understanding of the molecular and cellular mechanisms involved in its formation provides insights into congenital malformations affecting its function and therefore the survival of the organism. Because many developmental mechanisms are highly conserved, it is possible to extrapolate from observations made in invertebrate and vertebrate model organisms to humans. This review will highlight the contributions made through studying heart development in avian embryos, particularly the chicken. The major advantage of chick embryos is their accessibility for surgical manipulation and functional interference approaches, both gain- and loss-of-function. In addition to experiments performed in ovo, the dissection of tissues for ex vivo culture, genomic, or biochemical approaches is straightforward. Furthermore, embryos can be cultured for time-lapse imaging, which enables tracking of fluorescently labeled cells and detailed analysis of tissue morphogenesis. Owing to these features, investigations in chick embryos have led to important discoveries, often complementing genetic studies in mice and zebrafish. As well as including some historical aspects, we cover here some of the crucial advances made in understanding early heart development using the chicken model.

  19. Stage selection and restricted oviposition period improves cryopreservation of dipteran embryos.

    Science.gov (United States)

    Rajamohan, Arun; Rinehart, Joseph P; Leopold, Roger A

    2015-04-01

    Embryos of two dipteran species (Musca domestica and Lucilia sericata) were assessed for an effective sampling time that would result in the highest post-cryopreservation hatch rate, with a primary goal to define species-specific egg collection periods and the effects of manual stage selection on post cryopreservation yield. The effects of the time taken to collect eggs on, (a) the proportion of embryos reaching a specific developmental stage between 17 and 20 h of development, and (b) the post-cryopreservation hatch rate were assessed. Permeabilization treatment applied at any stage of embryonic development did not significantly reduce embryo viability. Eggs collected over longer durations significantly reduced the number of embryos available in a specific developmental stage amenable to cryopreservation. Hatch percentage after cryopreservation of the embryos of M. domestica collected over a 60 min period was 10.7 ± 8.7% compared to 31 ± 5% for the eggs collected for just 15 min. Similarly, percent hatch in L. sericata resulted in 17.0 ± 3.9 and cryopreservation after manual selection of specific embryonic developmental stages from the dechorionated samples. Post-cryopreservation hatching rate for stage-selected M. domestica embryos was 86.5 ± 5.5% compared to 33.3 ± 4.5% for embryos staged only by an overall visual confirmation. In the case of L. sericata, the hatching percentage was 79.0 ± 11.1 for stage-selected embryos compared to 17.0 ± 3.9% without individual selection. Published by Elsevier Inc.

  20. Halogenated flame retardants during egg formation and chicken embryo development: maternal transfer, possible biotransformation, and tissue distribution.

    Science.gov (United States)

    Zheng, Xiao-Bo; Luo, Xiao-Jun; Zeng, Yan-Hong; Wu, Jiang-Ping; Chen, She-Jun; Mai, Bi-Xian

    2014-08-01

    Hen muscle, eggs, and newborn chick tissues (muscle and liver) were collected from an electronic waste recycling site in southern China. The authors examined the maternal transfer, potential metabolism, and tissue distribution of several halogenated flame retardants (HFRs) during egg formation and chicken embryo development. The pollutant composition changes significantly from hen muscle to eggs and from eggs to tissues of newborn chicks. Higher-halogenated chemicals, such as octa- to deca-polybrominated diphenyl ether (PBDE) congeners, deca-polybrominated biphenyl (PBB209), and dechlorane plus (DP), are less readily transferred to eggs compared with lower-halogenated chemicals. During embryo development, PBDEs are the most likely to be metabolized, whereas decabromodiphenyl ethane (DBDPE) is the least. The authors also observed selective maternal transfer of anti-DP and stereoselective metabolism of syn-DP during chicken embryo development. During tissue development, liver has greater affinity than the muscle for chemcials with a high log octanol-water partition coefficient, with the exception of DBDPE. The differences in metabolism potential of different chemicals in chicken embryos cause pollutant composition alterations. Halogenated flame retardant from maternal transfer and tissue distribution also exhibited chemical specificity, especially for DBDPE. Levels of DBDPE were elevated along with the full process from hen muscle to eggs and from eggs to chick tissues. More attention should be paid to the selective accumulation and biotransformation of HFRs in the early development stage of birds. © 2014 SETAC.

  1. Embryo transfer simulation improves pregnancy rates and decreases time to proficiency in Reproductive Endocrinology and Infertility fellow embryo transfers.

    Science.gov (United States)

    Heitmann, Ryan J; Hill, Micah J; Csokmay, John M; Pilgrim, Justin; DeCherney, Alan H; Deering, Shad

    2017-05-01

    To design and evaluate an ET simulator to train Reproductive Endocrinology and Infertility (REI) fellows' techniques of ET. Simulation model development and retrospective cohort analysis. Not applicable. Patients undergoing IVF. Simulation model evaluation and implementation of ET simulation training. Pregnancy rates. The REI fellow and faculty evaluation responses (n = 19/21 [90%]) of the model demonstrated realistic characteristics, with evaluators concluding the model was suitable for training in almost all evaluated areas. A total of 12 REI fellows who performed ET were analyzed: 6 before ET trainer and 6 after ET trainer. Pregnancy rates were 31% in the initial 10 ETs per fellow before simulator vs. 46% after simulator. One of six pre-ET trainer fellows (17%) had pregnancy rates ≥40% in their first 10 ETs; whereas four of six post-ET trainer fellows had pregnancy rates ≥40% in their first 10 ETs. The average number of ETs to obtain >40% pregnancy efficiency was 27 ETs before trainer vs. 15 ETs after trainer. Pregnancy rates were similar in the two groups after 20 ETs, and collective terminal pregnancy rates were >50% after 40 ETs. Embryo transfer simulation improved REI fellow pregnancy rates in their first 10 transfers and led to a more rapid ET proficiency. These data suggest potential value in adopting ET simulation, even in programs with a robust history of live ET in fellowship training. Published by Elsevier Inc.

  2. State-of-the-art production, conservation and transfer of in-vitro-produced embryos in small ruminants.

    Science.gov (United States)

    Cognié, Yves; Poulin, Nati; Locatelli, Yann; Mermillod, Pascal

    2004-01-01

    Today, although not efficient enough to replace multiple ovulation and embryo transfer, in vitro embryo production for small ruminants is a platform for new reproductive technologies, such as embryo sexing, transgenesis and cloning. The in vitro embryo-production system developed for sheep and goats is more efficient now than 15 years ago, but could still be improved. Laparoscopic collection of oocytes in live animals treated with gonadotrophin indicates a promising future for the application of this technology to genetic improvement programmes. Oocyte maturation in defined medium with epidermal growth factor and cysteamine appears as efficient as oocyte maturation in follicular fluid-supplemented medium and allows future study of the effect of other factors involved in the cytoplasmic maturation of oocytes from these species. Further efforts have to be made to standardise the semen-capacitating process and to improve the quality and freezability of in-vitro-produced (IVP) embryos. The optimisation of IVP procedures for deer species has required the study of the seasonal variation of oocyte competence and the development of a specific methodology to allow the culture of embryos up to the blastocyst stage.

  3. A comparison of psychological functioning in couples undergoing frozen-thawed embryo replacement in various stages of treatment using the Mean Affect Adjective Check List (MAACL).

    Science.gov (United States)

    Ismail, W A; Menezes, M Q; Martin, C W; Thong, K J

    2004-09-01

    The objectives were to identify the stage(s) of frozen embryo replacement cycle where the couples are most vulnerable to psychological dysfunction. Assessment was performed by using the Mean Affect Adjective Check-List (MAACL). Thirty couples completed the MAACL questionnaire at the following stages: (a) pretreatment (visit 1), (b) before embryo transfer (visit 2), and (c) before pregnancy test (visit 3). Each partner had to complete a separate questionnaire set. For both partners, the depression score for visit 3 was significantly higher and the sensation seeking and positive affect scores were significantly lower than the corresponding scores for earlier visits. Anxiety scores were similar for all visits. For men, the hostility scores were significantly higher between visits 1 and 2 while in women the same was reported between visits 3 and 2. Psychological counselling should be targeted at couples especially after embryo transfer. MAACL is a useful method for measuring psychological dysfunction in these couples.

  4. Strain preservation of experimental animals: vitrification of two-cell stage embryos for multiple mouse strains.

    Science.gov (United States)

    Eto, Tomoo; Takahashi, Riichi; Kamisako, Tsutomu

    2015-04-01

    Strain preservation of experimental animals is crucial for experimental reproducibility. Maintaining complete animal strains, however, is costly and there is a risk for genetic mutations as well as complete loss due to disasters or illness. Therefore, the development of effective vitrification techniques for cryopreservation of multiple experimental animal strains is important. We examined whether a vitrification method using cryoprotectant solutions, P10 and PEPeS, is suitable for preservation of multiple inbred and outbred mouse strains. First, we investigated whether our vitrification method using cryoprotectant solutions was suitable for two-cell stage mouse embryos. In vitro development of embryos exposed to the cryoprotectant solutions was similar to that of fresh controls. Further, the survival rate of the vitrified embryos was extremely high (98.1%). Next, we collected and vitrified two-cell stage embryos of 14 mouse strains. The average number of embryos obtained from one female was 7.3-33.3. The survival rate of vitrified embryos ranged from 92.8% to 99.1%, with no significant differences among mouse strains. In vivo development did not differ significantly between fresh controls and vitrified embryos of each strain. For strain preservation using cryopreserved embryos, two offspring for inbred lines and one offspring for outbred lines must be produced from two-cell stage embryos collected from one female. The expected number of surviving fetuses obtained from embryos collected from one female of either the inbred or outbred strains ranged from 2.9 to 19.5. The findings of the present study indicated that this vitrification method is suitable for strain preservation of multiple mouse strains. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  5. Development of a generally applicable morphokinetic algorithm capable of predicting the implantation potential of embryos transferred on Day 3

    Science.gov (United States)

    Petersen, Bjørn Molt; Boel, Mikkel; Montag, Markus; Gardner, David K.

    2016-01-01

    curve (AUC) to establish the predictive strength of the algorithm. MAIN RESULTS AND THE ROLE OF CHANCE By applying the here developed algorithm (KIDScore), which was based on six annotations (the number of pronuclei equals 2 at the 1-cell stage, time from insemination to pronuclei fading at the 1-cell stage, time from insemination to the 2-cell stage, time from insemination to the 3-cell stage, time from insemination to the 5-cell stage and time from insemination to the 8-cell stage) and ranking the embryos in five groups, the implantation potential of the embryos was predicted with an AUC of 0.650. On Day 3 the KIDScore algorithm was capable of predicting blastocyst development with an AUC of 0.745 and blastocyst quality with an AUC of 0.679. In a comparison of blastocyst prediction including six other published algorithms and KIDScore, only KIDScore and one more algorithm surpassed an algorithm constructed on conventional Alpha/ESHRE consensus timings in terms of predictive power. LIMITATIONS, REASONS FOR CAUTION Some morphological assessments were not available and consequently three of the algorithms in the comparison were not used in full and may therefore have been put at a disadvantage. Algorithms based on implantation data from Day 3 embryo transfers require adjustments to be capable of predicting the implantation potential of Day 5 embryo transfers. The current study is restricted by its retrospective nature and absence of live birth information. Prospective Randomized Controlled Trials should be used in future studies to establish the value of time-lapse technology and morphokinetic evaluation. WIDER IMPLICATIONS OF THE FINDINGS Algorithms applicable to different culture conditions can be developed if based on large data sets of heterogeneous origin. STUDY FUNDING/COMPETING INTEREST(S) This study was funded by Vitrolife A/S, Denmark and Vitrolife AB, Sweden. B.M.P.’s company BMP Analytics is performing consultancy for Vitrolife A/S. M.B. is employed at

  6. Non-invasive metabolomic profiling of embryo culture media and morphology grading to predict implantation outcome in frozen-thawed embryo transfer cycles.

    Science.gov (United States)

    Li, Xiong; Xu, Yan; Fu, Jing; Zhang, Wen-Bi; Liu, Su-Ying; Sun, Xiao-Xi

    2015-11-01

    Assessment of embryo viability is a crucial component of in vitro fertilization and currently relies largely on embryo morphology and cleavage rate. Because morphological assessment remains highly subjective, it can be unreliable in predicting embryo viability. This study investigated the metabolomic profiling of embryo culture media using near-infrared (NIR) spectroscopy for predicting the implantation potential of human embryos in frozen-thawed embryo transfer (FET) cycles. Spent embryo culture media was collected on day 4 after thawed embryo transfer (n = 621) and analysed using NIR spectroscopy. Viability scores were calculated using a predictive multivariate algorithm of fresh embryos with known pregnancy outcomes. The mean viability indices of embryos resulting in clinical pregnancy following FET were significantly higher than those of non-implanted embryos and differed between the 0, 50, and 100 % implantation groups. Notably, the 0 % group index was significantly lower than the 100 % implantation group index (-0.787 ± 0.382 vs. 1.064 ± 0.331, P  0.05). NIR metabolomic profiling of thawed embryo culture media is independent of morphology and correlates with embryo implantation potential in FET cycles. The viability score alone or in conjunction with morphologic grading is a more objective marker for implantation outcome in FET cycles than morphology alone.

  7. [Intravaginal culture and embryo transfer. A new method for the fertilization of human oocytes].

    Science.gov (United States)

    Ranoux, C; Dubuisson, J B; Foulot, H; Aubriot, F X

    1987-12-01

    This technique was developed at the University Clinic of Port-Royal. It corresponds to the intravaginal culture of embryos and their transfer into the uterus. After ovocyte stimulation, most often by Clomid HMG, the follicles are aspirated under laparoscopic or sonographic control 34 to 36 hours after HCG. After being collected, the ovocytes are placed, whatever their stage of maturity, in one or several 3 ml tubes completely filled with culture medium (B2 of pure Menezo). Up to 4 ovocytes per tube are thus fertilized with 10 to 20,000 mobile spermatozoids/ml, prepared in the usual dilution, centrifugation and migration. Then the tube(s) are placed in the posterior vaginal cul-de-sac, kept in place with a diaphragm where they will remain during the 44 to 48 hours of culture time. Following that time, the contents of the tube are examined in order to evaluate the occurrence and the stage of embryonic division. A first series of 100 aspirations has enabled to obtain 15 pregnancies, still evolving, including two births of healthy children. A randomized series is currently in progress to determine a possible difference in the rates of pregnancy between CIVETE and the classic technique. Beside its new psychological contribution, this technique has demonstrated that it was possible to culture human embryos in the absence of CO2; its extreme simplicity should lead to a broader expansion of this technique.

  8. Dynamic distribution of NuMA and microtubules in human fetal fibroblasts, developing oocytes and somatic cell nuclear transferred embryos.

    Science.gov (United States)

    Xu, Xiaoming; Duan, Xin; Lu, Changfu; Lin, Ge; Lu, Guangxiu

    2011-05-01

    The nuclear mitotic apparatus (NuMA) plays a central role in the assembly and maintenance of spindle poles. Somatic cell nuclear transfer (SCNT) studies on non-human primates have shown that meiotic spindle removal during enucleation causes depletion of NuMA and the minus-end-directed motor protein (HSET) from the ooplasm, and this in turn leads to failure of embryo development. To determine whether NuMA from somatic cells could compensate for NuMA loss during enucleation, the distribution of NuMA and microtubule organization were investigated in human fibroblasts, developing oocytes and SCNT embryos. Human fetal fibroblasts, oocytes at various maturation stages and human embryos reconstructed by different SCNT methods were analyzed for NuMA and α-tubulin using immunofluorescent confocal microscopy. NuMA was detected in interphase nuclei of fibroblasts and oocytes. During mitosis and meiosis, NuMA relocated to the domain surrounding the two spindle poles. During the enucleation process, NuMA was removed along with the meiotic spindle. At 2 h after injection into a donor cell, transitory bipolar spindles were organized and NuMA was detected in the reformed poles. NuMA could be detected spreading uniformly across the nucleoplasm of one pseudo-pronucleus in SCNT embryos but was excluded from the nucleolus. Regardless of the method used for SCNT (enucleation-injection or injection-pronuclei enucleation), NuMA aggregated and relocated to the reformed spindle poles at metaphase of the first mitotic event. At interphase, NuMA relocated throughout the nucleus in developmentally arrested SCNT embryos. Our results show that donor cell nuclei contain NuMA, which might contribute to the maintenance of spindle morphology in SCNT embryos. Normal spindle and NuMA expression were found in human SCNT embryos at different developmental stages.

  9. Non-invasive assessment of in-vitro embryo quality to improve transfer success

    DEFF Research Database (Denmark)

    Højbøge, Tina Rødgaard; Heegaard, Peter M. H.; Callesen, Henrik

    2015-01-01

    embryos before the transfer to a recipient still remains challenging. Presently, the predominant non-invasive technique for selecting viable embryos is based on morphology, where parameters such as rates of cleavage and blastocyst formation as well as developmental kinetics are evaluated mostly...... subjectively. The simple morphological approach is, however, inadequate for the prediction of embryo quality, and several studies have focused on developing new non-invasive methods using molecular approaches based particularly on proteomics, metabolomics and most recently small non-coding RNA, including micro......RNA. This review outlines the potential of several non-invasive in-vitro methods based on analysis of spent embryo culture medium....

  10. Reprogrammed transcriptome in rhesus-bovine interspecies somatic cell nuclear transfer embryos.

    Directory of Open Access Journals (Sweden)

    Kai Wang

    Full Text Available Global activation of the embryonic genome (EGA, one of the most critical steps in early mammalian embryo development, is recognized as the time when interspecies somatic cell nuclear transfer (iSCNT embryos fail to thrive.In this study, we analyzed the EGA-related transcriptome of rhesus-bovine iSCNT 8- to 16-cell embryos and dissected the reprogramming process in terms of embryonic gene activation, somatic gene silencing, and maternal RNA degradation. Compared with fibroblast donor cells, two thousand and seven genes were activated in iSCNT embryos, one quarter of them reaching expression levels comparable to those found in in vitro fertilized (IVF rhesus embryos. This suggested that EGA in iSCNT embryos had partially recapitulated rhesus embryonic development. Eight hundred and sixty somatic genes were not silenced properly and continued to be expressed in iSCNT embryos, which indicated incomplete nuclear reprogramming. We compared maternal RNA degradation in bovine oocytes between bovine-bovine SCNT and iSCNT embryos. While maternal RNA degradation occurred in both SCNT and iSCNT embryos, we saw more limited overall degradation of maternal RNA in iSCNT embryos than in SCNT embryos. Several important maternal RNAs, like GPF9, were not properly processed in SCNT embryos.Our data suggested that iSCNT embryos are capable of triggering EGA, while a portion of somatic cell-associated genes maintain their expression. Maternal RNA degradation seems to be impaired in iSCNT embryos. Further understanding of the biological roles of these genes, networks, and pathways revealed by iSCNT may expand our knowledge about cell reprogramming, pluripotency, and differentiation.

  11. Artificial cryopreserved embryo transfer cycle success depends on blastocyst developmental rate and progesterone timing

    DEFF Research Database (Denmark)

    Ozgur, Kemal; Bulut, Hasan; Berkkanoglu, Murat

    2018-01-01

    This retrospective cohort analysis compared the developmental competence of cryopreserved day-4 and 5 blastocysts, and investigated the effect of progesterone administration duration on the success of artificial frozen embryo transfers. Between October 2015 and March 2016, 868 intracytoplasmic sp...

  12. Reducing twin pregnancy rates after IVF--elective single embryo transfer (eSET).

    LENUS (Irish Health Repository)

    Milne, P

    2010-01-01

    Multiple pregnancy is a major complication of IVF and is associated with increased maternal, fetal and neonatal morbidity. Elective single embryo transfer (eSET) during IVF, rather than the more standard transfer of two embryos (double embryo transfer or DET), has been shown to significantly reduce the multiple pregnancy rate associated with IVF, while maintaining acceptable pregnancy rates. Couples undergoing IVF in 2008 who met good prognostic criteria had eSET performed. Pregnancy and twinning rates were compared with those for similar couples in 2007 who had DET. Couples unsuccessful with a fresh cycle of treatment had subsequent frozen embryo transfer cycles with DET. The cumulative pregnancy rate was similar for each group. However there were no multiple pregnancies in the eSET group, compared to 4 twins of 5 pregnancies in the DET group. 96% of eligible couples agreed to eSET. ESET is successful in and acceptable to good prognosis Irish couples undergoing IVF.

  13. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF.

    Science.gov (United States)

    Vergouw, Carlijn G; Kostelijk, E Hanna; Doejaaren, Els; Hompes, Peter G A; Lambalk, Cornelis B; Schats, Roel

    2012-09-01

    Does the type of medium used to culture fresh and frozen-thawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean birthweight adjusted for gestational age, gender and parity (z-scores) of singletons born after a fresh or frozen-thawed SET. Furthermore, we show that embryo freezing and thawing cycles may lead to a significantly higher mean birthweight. Animal studies have shown that culture media constituents are responsible for changes in birthweight of offspring. In human IVF, there is still little knowledge of the effect of medium type on birthweight. Until now, only a small number of commercially available culture media have been investigated (Vitrolife, Cook(®) Medical and IVF online medium). Our study adds new information: it has a larger population of singleton births compared with the previously published studies, it includes outcomes of other media types (HTF and Sage(®)), not previously analysed, and it includes data on frozen-thawed SETs. This study was a retrospective analysis of birthweights of singleton newborns after fresh (Day 3) or frozen-thawed (Day 5) SET cycles, using embryos cultured in either of two different types of commercially available culture media, between 2008 and 2011. Before January 2009, a single-step culture medium was used: human tubal fluid (HTF) with 4 mg/ml human serum albumin. From January 2009 onwards, a commercially available sequential medium was introduced: Sage(®), Quinn's advantage protein plus medium. Singletons born after a fresh SET (99 embryos cultured in HTF and 259 in Sage(®)) and singletons born after a frozen-thawed SET (32 embryos cultured in HTF only, 41 in HTF and Sage(®) and 86 in Sage(®) only) were analysed. Only patients using autologous gametes without the use of a gestational carrier were considered. Also excluded were (vanishing) twins, triplets

  14. Pentachlorophenol exposure causes Warburg-like effects in zebrafish embryos at gastrulation stage

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Ting; Zhao, Jing [Key Laboratory of Yangtze River Water Environment, Ministry of Education, College of Environmental Science and Technology, Tongji University, Shanghai 200092 (China); Hu, Ping [Key Laboratory of Model Animal for Disease Study, Ministry of Education, Model Animal Research Center, Nanjing University, Nanjing 210061 (China); State Key Laboratory of Reproductive Medicine, Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029 (China); Dong, Zhangji; Li, Jingyun [Key Laboratory of Model Animal for Disease Study, Ministry of Education, Model Animal Research Center, Nanjing University, Nanjing 210061 (China); Zhang, Hongchang [Key Laboratory of Yangtze River Water Environment, Ministry of Education, College of Environmental Science and Technology, Tongji University, Shanghai 200092 (China); Yin, Daqiang, E-mail: yindq@tongji.edu.cn [Key Laboratory of Yangtze River Water Environment, Ministry of Education, College of Environmental Science and Technology, Tongji University, Shanghai 200092 (China); Zhao, Qingshun, E-mail: qingshun@nju.edu.cn [Key Laboratory of Model Animal for Disease Study, Ministry of Education, Model Animal Research Center, Nanjing University, Nanjing 210061 (China)

    2014-06-01

    Pentachlorophenol (PCP) is a prevalent pollutant in the environment and has been demonstrated to be a serious toxicant to humans and animals. However, little is known regarding the molecular mechanism underlying its toxic effects on vertebrate early development. To explore the impacts and underlying mechanisms of PCP on early development, zebrafish (Danio rerio) embryos were exposed to PCP at concentrations of 0, 20 and 50 μg/L, and microscopic observation and cDNA microarray analysis were subsequently conducted at gastrulation stage. The morphological observations revealed that PCP caused a developmental delay of zebrafish embryos in a concentration-dependent manner. Transcriptomic data showed that 50 μg/L PCP treatment resulted in significant changes in gene expression level, and the genes involved in energy metabolism and cell behavior were identified based on gene functional enrichment analysis. The energy production of embryos was influenced by PCP via the activation of glycolysis along with the inhibition of oxidative phosphorylation (OXPHOS). The results suggested that PCP acts as an inhibitor of OXPHOS at 8 hpf (hours postfertilization). Consistent with the activated glycolysis, the cell cycle activity of PCP-treated embryos was higher than the controls. These characteristics are similar to the Warburg effect, which occurs in human tumors. The microinjection of exogenous ATP confirmed that an additional energy supply could rescue PCP-treated embryos from the developmental delay due to the energy deficit. Taken together, our results demonstrated that PCP causes a Warburg-like effect on zebrafish embryos during gastrulation, and the affected embryos had the phenotype of developmental delay. - Highlights: • We treat zebrafish embryos with PCP at gastrula stage. • PCP acts as an oxidative phosphorylation inhibitor, not an uncoupler, in gastrulation. • Exogenous ATP injection will rescue the development of effected embryos. • The transcriptome of PCP

  15. Pentachlorophenol exposure causes Warburg-like effects in zebrafish embryos at gastrulation stage

    International Nuclear Information System (INIS)

    Xu, Ting; Zhao, Jing; Hu, Ping; Dong, Zhangji; Li, Jingyun; Zhang, Hongchang; Yin, Daqiang; Zhao, Qingshun

    2014-01-01

    Pentachlorophenol (PCP) is a prevalent pollutant in the environment and has been demonstrated to be a serious toxicant to humans and animals. However, little is known regarding the molecular mechanism underlying its toxic effects on vertebrate early development. To explore the impacts and underlying mechanisms of PCP on early development, zebrafish (Danio rerio) embryos were exposed to PCP at concentrations of 0, 20 and 50 μg/L, and microscopic observation and cDNA microarray analysis were subsequently conducted at gastrulation stage. The morphological observations revealed that PCP caused a developmental delay of zebrafish embryos in a concentration-dependent manner. Transcriptomic data showed that 50 μg/L PCP treatment resulted in significant changes in gene expression level, and the genes involved in energy metabolism and cell behavior were identified based on gene functional enrichment analysis. The energy production of embryos was influenced by PCP via the activation of glycolysis along with the inhibition of oxidative phosphorylation (OXPHOS). The results suggested that PCP acts as an inhibitor of OXPHOS at 8 hpf (hours postfertilization). Consistent with the activated glycolysis, the cell cycle activity of PCP-treated embryos was higher than the controls. These characteristics are similar to the Warburg effect, which occurs in human tumors. The microinjection of exogenous ATP confirmed that an additional energy supply could rescue PCP-treated embryos from the developmental delay due to the energy deficit. Taken together, our results demonstrated that PCP causes a Warburg-like effect on zebrafish embryos during gastrulation, and the affected embryos had the phenotype of developmental delay. - Highlights: • We treat zebrafish embryos with PCP at gastrula stage. • PCP acts as an oxidative phosphorylation inhibitor, not an uncoupler, in gastrulation. • Exogenous ATP injection will rescue the development of effected embryos. • The transcriptome of PCP

  16. Large baby syndrome in singletons born after frozen embryo transfer (FET)

    DEFF Research Database (Denmark)

    Pinborg, Anja; Henningsen, AA; Loft, A

    2013-01-01

    Are singletons born after frozen embryo transfer (FET) at increased risk of being born large for gestational age (LGA) and if so, is this caused by intrinsic maternal factors or related to the freezing/thawing procedures?......Are singletons born after frozen embryo transfer (FET) at increased risk of being born large for gestational age (LGA) and if so, is this caused by intrinsic maternal factors or related to the freezing/thawing procedures?...

  17. An investigation into the possibility of bluetongue virus transmission by transfer of infected ovine embryos

    Directory of Open Access Journals (Sweden)

    Estelle H. Venter

    2011-02-01

    Full Text Available Bluetongue (BT, a disease that affects mainly sheep, causes economic losses owing to not only its deleterious effects on animals but also its associated impact on the restriction of movement of livestock and livestock germplasm. The causative agent, bluetongue virus (BTV, can occur in the semen of rams and bulls at the time of peak viraemia and be transferred to a developing foetus. The risk of the transmission of BTV by bovine embryos is negligible if the embryos are washed according to the International Embryo Transfer Society (IETS protocol. Two experiments were undertaken to determine whether this holds for ovine embryos that had been exposed to BTV. Firstly, the oestrus cycles of 12 ewes were synchronised and the 59 embryos that were obtained were exposed in vitro to BTV-2 and BTV-4 at a dilution of 1 x 102.88 and 1 x 103.5 respectively. In the second experiment, embryos were recovered from sheep at the peak of viraemia. A total of 96 embryos were collected from BTV-infected sheep 21 days after infection. In both experiments half the embryos were washed and treated with trypsin according to the IETS protocol while the remaining embryos were neither washed nor treated. All were tested for the presence of BTV using cell culture techniques. The virus was detected after three passages in BHK-21 cells only in one wash bath in the first experiment and two unwashed embryos exposed to BTV-4 at a titre of 1 x 103.5. No embryos or uterine flush fluids obtained from viraemic donors used in the second experiment were positive for BTV after the standard washing procedure had been followed. The washing procedure of the IETS protocol can thus clear sheep embryos infected with BTV either in vitro or in vivo.

  18. The Relationship between Cell Number, Division Behavior and Developmental Potential of Cleavage Stage Human Embryos: A Time-Lapse Study.

    Directory of Open Access Journals (Sweden)

    Xiangyi Kong

    Full Text Available Day 3 cleavage embryo transfer is routine in many assisted reproductive technology centers today. Embryos are usually selected according to cell number, cell symmetry and fragmentation for transfer. Many studies have showed the relationship between cell number and embryo developmental potential. However, there is limited understanding of embryo division behavior and their association with embryo cell number and developmental potential. A retrospective and observational study was conducted to investigate how different division behaviors affect cell number and developmental potential of day 3 embryos by time-lapse imaging. Based on cell number at day 3, the embryos (from 104 IVF/intracytoplasmic sperm injection (ICSI treatment cycles, n = 799 were classified as follows: less than 5 cells (10C; n = 42. Division behavior, morphokinetic parameters and blastocyst formation rate were analyzed in 5 groups of day 3 embryos with different cell numbers. In 10C embryos increased compared to 7-8C embryos (45.8%, 33.3% vs. 11.1%, respectively. In ≥5C embryos, FR and DC significantly reduced developmental potential, whereas 10C. In NB embryos, the cell cycle elongation or shortening was the main cause for abnormally low or high cell number, respectively. After excluding embryos with abnormal division behaviors, the developmental potential, implantation rate and live birth rate of day 3 embryos increased with cell number.

  19. Employing mated females as recipients for transfer of cloned dog embryos.

    Science.gov (United States)

    Kim, Geon A; Oh, Hyun Ju; Park, Jung Eun; Kim, Min Jung; Park, Eun Jung; Lim, Sang Hyun; Kang, Sung Keun; Jang, Goo; Lee, Byeong Chun

    2013-01-01

    It has been suggested that co-transferring parthenogenetic embryos could improve the pregnancy success rate with cloned embryos in mammals. As an alternative to co-transferring parthenotes, in dogs we employed recipient females that possessed in vivo-fertilised embryos as a result of mating to determine whether mated bitches could be suitable recipients for cloned embryos. The effect of using mated recipients on implantation and pregnancy rates of canine somatic cell nuclear transfer embryos was also determined. Cloned embryos were transferred into the oviducts of naturally synchronous females that had mated with male dogs before ovulation. The pregnancy rate appeared to be similar between mated recipients (50%) and non-mated recipients (28.57%; P>0.05). However, the delivery rate of cloned pups was significantly higher in mated recipients than non-mated recipients (10.53 vs 2.38%; Pcloned pups in non-mated recipients were delivered by Caesarean section because the fall in progesterone concentration in these females did not occur until the due date. The present study demonstrated for the first time that mated female dogs can be used as recipients for cloned embryos.

  20. Repeated embryo collection and interspecies transfer in alpacas and llamas during non-breeding season

    Directory of Open Access Journals (Sweden)

    Pacheco J

    2015-08-01

    Full Text Available Sexual behavior evaluation was evaluated, collecting and interspecies embryo transfer inter species in llamas and alpacas during non-breeding season, 10 and 10 donor alpacas llamas, alpacas and 20 receiving 20 llamas, 5 alpacas and 5 llamas males were used. Sexual behavior by libido in males and acceptance of female to male in the presence of a dominant follicle was evaluated, the collection of embryos simple ovulation by non-surgical technique was performed and the fresh embryos are transferred directly into the horn left. It was observed that only 40% of alpaca accept the male and female in all cases had to use two males for mating, but all llama males mounted on the first attempt and accepted all females breeding. Embryos were collected at 25 and 60% of alpacas and llamas washes respectively, all were grade 1 embryos transferable; the embryo transfer fertility evaluated by ultrasound at 25 days was 100 and 41.6% respectively for donor alpaca and llama, however ultrasound evaluation at 60 days fertility was 50 and 25% respectively for donor alpaca and llama. We conclude that there is greater reproductive seasonality in alpaca regard to llamas, all were grade 1 embryos collected, fertility evaluated by ultrasound 25 days down to 60 days, demonstrating embryonic mortality, possibly due to the non-breeding season of both species.

  1. Interspecific embryo transfer from mouflon (Ovis gmelini musimon) to domestic Corriedale sheep in Argentina.

    Science.gov (United States)

    Dixon, T E; Hunter, J W; Soler, J P

    2007-09-01

    An interspecific embryo transfer program was conducted for genetic improvement and increasing the number of offspring from a flock of mouflon sheep in Argentina. The female donor mouflons were divided into three groups, G1 (n=5), G2 (n=4) and G3 (n=5). The total NIH-FSH-P1 dose given to each donor on the superovulatory treatment was 260, 200 and 160 mg for G1, G2 and G3, respectively. The mouflons in G3 were maidens, while the others were multiparous. Domestic Corriedale ewes (n=60) were synchronized and used as recipients. The embryo recovery and transfer was performed by a surgical method. Mouflons (n=13) responded to the superovulatory treatment with an average of 9.1+/-2.8 ovulations. A low incidence of early luteal regression was found (1 out of 14 donors). Embryo recovery rates were 60, 31 and 76% in groups G1, G2 and G3, respectively. The percentage of transferable embryos obtained in G1 and in G2 exceeded 80%. None of the embryos obtained from G3 were of transferable quality. In G1, 25 transferable embryos were recovered and transferred to 13 recipients, resulting in a pregnancy rate of 76.9% (10/13). In G2, 10 embryos were transferred to 5 recipients, resulting in a 60% pregnancy rate (3/5). Lambing rate was 60% (15/25) and 30% (3/10) for G1 and G2, respectively. Thirteen lambs were born to the 14 donors following natural service after the embryo recoveries. This study demonstrates that the application of IET technology would have great reproductive impact, especially when the donor mouflon hinds are selected according to age and reproductive history.

  2. Vitamin C supplementation enhances compact morulae formation but reduces the hatching blastocyst rate of bovine somatic cell nuclear transfer embryos.

    Science.gov (United States)

    Li, Qian; Wang, Yong-Sheng; Wang, Li-Jun; Zhang, Hui; Li, Rui-Zhe; Cui, Chen-Chen; Li, Wen-Zhe; Zhang, Yong; Jin, Ya-Ping

    2014-08-01

    Vitamin C, an antioxidant that reduces reactive oxygen species (ROS) in cells, is capable of significantly improving the developmental competence of porcine and mouse somatic cell nuclear transfer (SCNT) embryos, both in vitro and in vivo. In the present study, the effects of vitamin C on the developmental competence of bovine SCNT embryos were investigated. The results indicated that vitamin C (40 μg/mL) positively affected the scavenging of intracellular ROS, cleavage rate at 24 h (76.67 vs. 68.26%, pvitamin C supplementation did not significantly affect the blastocyst formation rate and proportion of inner cell mass over total cells per blastocyst on day 7. Moreover, vitamin C supplementation obviously impaired the total cell numbers per blastocyst (97.20 ± 11.35 vs. 88.57 ± 10.43, pVitamin C supplementation preferentially improved the viability of bovine SCNT embryos prior to the blastocyst stage, but did not enhance the formation and quality of blastocysts in vitro. In conclusion, the effect of vitamin C on the development of bovine SCNT embryos is complex, and vitamin C is not a suitable antioxidant chemical for the in vitro culture of bovine SCNT embryos.

  3. Activation of ribosomal RNA genes in porcine embryos produced in vitro or by somatic cell nuclear transfer

    DEFF Research Database (Denmark)

    Bjerregaard, Bolette; Pedersen, Hanne Gervi; Jakobsen, Anne Sørig

    2007-01-01

    equal proportions of transcriptionally active and inactive embryos and essentially all embryos that developed to the 16-cell stage (n = 21) and further to the blastocyst stage (n = 19) contained only transcriptionally active cells. In conclusion, porcine embryos produced in vitro had an asynchronous...

  4. Temporal and developmental-stage variation in the occurrence of mitotic errors in tripronuclear human preimplantation embryos

    NARCIS (Netherlands)

    Mantikou, Eleni; van Echten-Arends, Jannie; Sikkema-Raddatz, Birgit; van der Veen, Fulco; Repping, Sjoerd; Mastenbroek, Sebastiaan

    2013-01-01

    Mitotic errors during early development of human preimplantation embryos are common, rendering a large proportion of embryos chromosomally mosaic. It is also known that the percentage of diploid cells in human diploid-aneuploid mosaic embryos is higher at the blastocyst than at the cleavage stage.

  5. Temporal and Developmental-Stage Variation in the Occurrence of Mitotic Errors in Tripronuclear Human Preimplantation Embryos

    NARCIS (Netherlands)

    Mantikou, Eleni; van Echten-Arends, Jannie; Sikkema-Raddatz, Birgit; van der Veen, Fulco; Repping, Sjoerd; Mastenbroek, Sebastiaan

    Mitotic errors during early development of human preimplantation embryos are common, rendering a large proportion of embryos chromosomally mosaic. It is also known that the percentage of diploid cells in human diploid-aneuploid mosaic embryos is higher at the blastocyst than at the cleavage stage.

  6. Chromosome fragility at FRAXA in human cleavage stage embryos at risk for fragile X syndrome.

    Science.gov (United States)

    Verdyck, Pieter; Berckmoes, Veerle; De Vos, Anick; Verpoest, Willem; Liebaers, Inge; Bonduelle, Maryse; De Rycke, Martine

    2015-10-01

    Fragile X syndrome (FXS), the most common inherited intellectual disability syndrome, is caused by expansion and hypermethylation of the CGG repeat in the 5' UTR of the FMR1 gene. This expanded repeat, also known as the rare fragile site FRAXA, causes X chromosome fragility in cultured cells from patients but only when induced by perturbing pyrimidine synthesis. We performed preimplantation genetic diagnosis (PGD) on 595 blastomeres biopsied from 442 cleavage stage embryos at risk for FXS using short tandem repeat (STR) markers. In six blastomeres, from five embryos an incomplete haplotype was observed with loss of all alleles telomeric to the CGG repeat. In all five embryos, the incomplete haplotype corresponded to the haplotype carrying the CGG repeat expansion. Subsequent analysis of additional blastomeres from three embryos by array comparative genomic hybridization (aCGH) confirmed the presence of a terminal deletion with a breakpoint close to the CGG repeat in two blastomeres from one embryo. A blastomere from another embryo showed the complementary duplication. We conclude that a CGG repeat expansion at FRAXA causes X chromosome fragility in early human IVF embryos at risk for FXS. © 2015 Wiley Periodicals, Inc.

  7. Embryo-transfer twinning and performance efficiency in beef production.

    Science.gov (United States)

    Guerra-Martinez, P; Dickerson, G E; Anderson, G B; Green, R D

    1990-12-01

    Effects of twinning on efficiency of beef production were estimated from results of bilateral transfer of two Angus x Hereford (AxH) embryos into each of 241 heifers and 84 cows (H, A, HxA or Holstein x H) over 4 yr. Calves were weaned at 180 d and fed either 220 d in a feedlot (1977) or 170 d on forage and 140 d in a feedlot (1978 to 1980). Effects of parity, twinning and sex of calf were estimated as covariates within year-breed of dam. Pregnancy at 45 to 60 d of gestation was 68% in heifers (H) and 74% in cows (C), with 40% single (S) and 60% twin (T) births. Dystocia was 28% in H vs 10% in C (P less than .05), and tended to be less (P greater than .05) for T than S in H. More placentas were retained (P less than .05) for T than for S in both H (35 vs 12%) and C (24 vs 4%). Twin gestations averaged 3 d shorter and subsequent calving intervals 13 d longer (P less than .05), but total calf mortality was slightly higher (P greater than .05). Abortions were 4% in H only. Twinning females lost maternal weight during late gestation (P less than .05) when crowding limited voluntary feed intake, while fetal requirements were 60% higher (P less than .01). Twins increased milk output 25% (P less than .05), but 11% higher feed intake maintained cow weight during lactation. Twinning reduced birth weight 13% and weaning weight 17% (P less than .05), but 400-d feedlot weight only 9% because of compensating feedlot gain. Twins gained 18% faster than S during postweaning 170-d forage feeding, but 5% slower in feedlot to 8% lighter 490-d weight (P less than .05). Assuming 40% higher veterinary and labor costs for twins, estimated integrated herd costs per unit of age-constant output value would be lower for T than for S production by about 24% for marketing either at weaning or at 400 d.

  8. DOT1L inhibitor improves early development of porcine somatic cell nuclear transfer embryos

    DEFF Research Database (Denmark)

    Tao, Jia; Zhang, Yu; Zuo, Xiaoyuan

    2017-01-01

    inhibitor EPZ004777 (EPZ), significantly improved reprogramming efficiency during the generation of mouse induced pluripotent stem cells. However, the roles of DOT1L in porcine nuclear transfer-mediated cellular reprogramming are not yet known. Here we showed that DOT1L inhibition via 0.5 nM EPZ treatment......Incomplete epigenetic reprogramming of the genome of donor cells causes poor early and full-term developmental efficiency of somatic cell nuclear transfer (SCNT) embryos. Previous research indicate that inhibition of the histone H3 K79 methyltransferase DOT1L, using a selective pharmacological...... for 12 or 24 h significantly enhanced the blastocyst rate of SCNT embryos and dramatically reduced the level of H3K79me2 during SCNT 1-cell embryonic development. Additionally, H3K79me2 level in the EPZ-treated SCNT embryos was similar to that in in vitro fertilized embryos, suggesting that DOT1L...

  9. Evidence of Placental Autophagy during Early Pregnancy after Transfer of In Vitro Produced (IVP) Sheep Embryos.

    Science.gov (United States)

    Toschi, Paola; Czernik, Marta; Zacchini, Federica; Fidanza, Antonella; Loi, Pasqualino; Ptak, Grażyna Ewa

    2016-01-01

    Pregnancies obtained by Assisted Reproductive Technologies (ART) are associated with limited maternal nutrient uptake. Our previous studies shown that in vitro culture of sheep embryos is associated with vascularization defects in their placentae and consequent reduction of embryo growth. Autophagy is a pro-survival cellular mechanism triggered by nutrient insufficiency. Therefore, the goal of our present study was to determine if autophagy is involved in early placental development after transfer of in vitro produced (IVP) embryos. To do this, placentae obtained following transfer of IVP sheep embryos were compared with placentae obtained after natural mating (control-CTR). The placentae were collected on day 20 post-fertilization and post-mating, respectively, and were analyzed using molecular (qPCR), ultrastructural and histological/immunological approaches. Our results show drastically increased autophagy in IVP placentae: high levels of expression (pembryos.

  10. Ultrasonographic fetal sex identification in pregnant sheep derived from natural mating and embryo transfer.

    Science.gov (United States)

    de Freitas Neto, Leopoldo Mayer; dos Santos, Maico Henrique Barbosa; de Aguiar Filho, Cristiano Rocha; de Almeida Irmão, José Monteiro; Caldas, Eduardo Luiz Cavalcanti; Neves, Jairo Pereira; Lima, Paulo Fernandes; de Oliveira, Marcos Antonio Lemos

    2010-06-01

    The objective of this study was to identify the migration period of the genital tubercle and the period of visualization of external genital structures in fetuses of the Dorper breed of sheep derived from natural mating and from fresh, frozen and vitrified embryo transfer. Transrectal ultrasound was performed using a double-frequency linear transducer (6.0 and 8.0 MHz) to monitor 130 ewe fetuses distributed in the four treatments regarding embryo origin. The accuracy of the ultrasound was 100% in this experiment. The fetuses originated from controlled natural mating (NM) and from fresh (FrE), frozen (FE) and vitrified (VE) embryo transfer, with embryos collected 7 days after breeding. Migration of the genital tubercle occurred earlier (Pembryos. It can also be concluded that real-time ultrasonography is a reliable tool for fetal sex determination in sheep taking into account both the location of the genital tubercle and the identification of external genital structures.

  11. Stage-dependent toxicity of bisphenol a on Rhinella arenarum (anura, bufonidae) embryos and larvae.

    Science.gov (United States)

    Wolkowicz, Ianina R Hutler; Herkovits, Jorge; Pérez Coll, Cristina S

    2014-02-01

    The acute and chronic toxicity of bisphenol A (BPA) was evaluated on the common South American toad Rhinella arenarum embryos and larvae by means of continuous and pulse exposure treatments. Embryos were treated continuously from early blastula (S.4) up to complete operculum (S.25), during early larval stages and by means of 24 h pulse exposures of BPA in concentrations ranging between 1.25 and 40 mg L(-1) , in order to evaluate the susceptibility to this compound in different developmental stages. For lethal effects, S.25 was the most sensitive and gastrula was the most resistant to BPA. The Teratogenic Index for neurula, the most sensitive embryonic stage for sublethal effects was 4.7. The main morphological alterations during early stages were: delayed or arrested development, reduced body size, persistent yolk plug, microcephaly, axial/tail flexures, edemas, blisters, waving fin, underdeveloped gills, mouth malformations, and cellular dissociation. BPA caused a remarkable narcotic effect from gill circulation stage (S.20) onwards in all the organisms exposed after 3 h of treatment with 10 mg L(-1) BPA. After recovering, the embryos exhibited scarce response to stimuli, erratic or circular swimming, and spasmodic contractions from 5 mg L(-1) onwards. Our results highlight the lethal and sublethal effectsof BPA on R. arenarum embryos and larvae, in the last case both at structural and functional levels. Copyright © 2011 Wiley Periodicals, Inc., A Wiley Company.

  12. Increasing twinning rate in beef cattle through embryo transfer ...

    African Journals Online (AJOL)

    Thirty-nine Limousin X Friesian heifers and forty-five Limousin X Friesian cows were randomly allocated to three treatments in a twin-induction breeding program. Animals were inseminated twice with Limousin semen (treatment A) or inseminated twice also with Hereford semen and each implanted with one embryo 7 days ...

  13. Coexpression analysis identifies nuclear reprogramming barriers of somatic cell nuclear transfer embryos.

    Science.gov (United States)

    Zuo, Yongchun; Su, Guanghua; Cheng, Lei; Liu, Kun; Feng, Yu; Wei, Zhuying; Bai, Chunling; Cao, Guifang; Li, Guangpeng

    2017-09-12

    The success of cloned animal "Dolly Sheep" demonstrated the somatic cell nuclear transfer (SCNT) technique holds huge potentials for mammalian asexual reproduction. However, the extremely poor development of SCNT embryos indicates their molecular mechanism remain largely unexplored. Deciphering the spatiotemporal patterns of gene expression in SCNT embryos is a crucial step toward understanding the mechanisms associated with nuclear reprogramming. In this study, a valuable transcriptome recourse of SCNT embryos was firstly established, which derived from different inter-/intra donor cells. The gene co-expression analysis identified 26 cell-specific modules, and a series of regulatory pathways related to reprogramming barriers were further enriched. Compared to the intra-SCNT embryos, the inter-SCNT embryos underwent only complete partially reprogramming. As master genome trigger genes, the transcripts related to TFIID subunit, RNA polymerase and mediators were incomplete activated in inter-SCNT embryos. The inter-SCNT embryos only wasted the stored maternal mRNA of master regulators, but failed to activate their self-sustained pathway of RNA polymerases. The KDM family of epigenetic regulator also seriously delayed in inter-SCNT embryo reprogramming process. Our study provided new insight into understanding of the mechanisms of nuclear reprogramming.

  14. Perinatal outcomes among singletons after assisted reproductive technology with single-embryo or double-embryo transfer versus no assisted reproductive technology.

    Science.gov (United States)

    Martin, Angela S; Chang, Jeani; Zhang, Yujia; Kawwass, Jennifer F; Boulet, Sheree L; McKane, Patricia; Bernson, Dana; Kissin, Dmitry M; Jamieson, Denise J

    2017-04-01

    To examine outcomes of singleton pregnancies conceived without assisted reproductive technology (non-ART) compared with singletons conceived with ART by elective single-embryo transfer (eSET), nonelective single-embryo transfer (non-eSET), and double-embryo transfer with the establishment of 1 (DET -1) or ≥2 (DET ≥2) early fetal heartbeats. Retrospective cohort using linked ART surveillance data and vital records from Florida, Massachusetts, Michigan, and Connecticut. Not applicable. Singleton live-born infants. None. Preterm birth (PTB Reproductive Medicine. All rights reserved.

  15. Timing of first embryonic cleavage is a positive indicator of the in vitro developmental potential of porcine embryos derived from in vitro fertilization, somatic cell nuclear transfer and parthenogenesis.

    Science.gov (United States)

    Isom, S Clay; Li, Rong Feng; Whitworth, Kristin M; Prather, Randall S

    2012-03-01

    Evidence in many species has suggested that those embryos that cleave earliest after fertilization are more developmentally competent than those that cleave relatively later after fertilization. Herein we document this phenomenon in porcine in vitro-fertilized (IVF), somatic cell nuclear transfer (SCNT), and parthenogenetic (PA) embryos. In vitro-matured pig oocytes were used to generate IVF, SCNT, and PA embryos. At 24 hr post-activation (or insemination; hpa/hpi), embryos were visually assessed, and cleaved embryos were moved into a new culture well. This process was repeated at 30 and 48 hpa/hpi. All embryos were allowed to develop 7 days in culture. For IVF embryos, 39.9%, 24.6%, and 10.5% of fast-, intermediate-, or slow-cleaving embryos, respectively, developed into blastocysts by day 7. For SCNT embryos, 31.8% of fast-, 5.7% of intermediate-, and 2.9% of late-cleaving embryos achieved the blastocyst stage of development. For PA embryos, the percentages of those cleaved embryos that developed to blastocyst were 59.3%, 36.7%, and 7.5% for early-, intermediate-, and late-cleaving embryos, respectively. Using RNA collected from early-, intermediate-, and late-cleaving embryos, real-time PCR was performed to assess the transcript levels of 14 different genes of widely varied function. The qPCR results suggest that maternal mRNA degradation may not proceed in an appropriate pattern in slow-cleaving embryos. These findings (1) confirm that, as observed in other species, earlier-cleaving porcine embryos are more successful at developing in culture than are slower-cleaving embryos, and (2) implicate mechanisms of maternal transcript destruction as potential determinants of oocyte/embryo quality. Copyright © 2011 Wiley Periodicals, Inc.

  16. Absence of Sperm Factors as in the Parthenogenesis Does Not Interfere on Bovine Embryo Sensitiveness to Heat Shock at Pre-Implantation Stage.

    Science.gov (United States)

    Camargo, L S A; Paludo, F; Pereira, M M; Wohlres-Viana, S; Gioso, M M; Carvalho, B C; Quintao, C C R; Viana, J H M

    2016-02-01

    Oocyte has been considered the major contributor for embryo thermo-tolerance. However, it was shown that sperm factors can be transferred to the oocyte during fertilization, raising the question of whether the absence of such factors could interfere on embryo thermo-tolerance. In this study, we used parthenogenesis to generate bovine embryos without spermatozoa in order to test whether the absence of sperm factors could influence their thermo-sensitiveness at early stages. In vitro fertilized (IVF) and parthenogenetic (PA) embryos at 44 h post-insemination/chemical activation were exposed to 38.5°C (control) or 41°C (heat shock) for 12 h and then developed for 48 h and up to blastocyst stage. Apoptosis index and expression of PRDX1, GLUT1, GLUT5 and IGF1r genes in blastocysts derived from heat-shocked embryos were also evaluated. The heat shock decreased the blastocyst rate at day seven (p 0.05) between method of activation (IVF and PA) and temperature (38.5°C or 41.5°C) for all developmental parameters evaluated. Expression of GLUT1 gene was downregulated (p < 0.05) by heat shock in both IVF and PA blastocyst whereas expression of GLUT5 and IGF1r genes was downregulated (p < 0.05) by heat shock in PA blastocysts. Those data show that the heat shock affects negatively the embryo development towards blastocysts stage, increases the apoptotic index and disturbed the expression of some genes in both IVF and PA embryos, indicating that the presence or absence of sperm factors does not influence the sensitivity of the bovine embryo to heat shock. © 2015 Blackwell Verlag GmbH.

  17. Embryo transfer practices in the United States: a survey of clinics registered with the Society for Assisted Reproductive Technology.

    Science.gov (United States)

    Jungheim, Emily S; Ryan, Ginny L; Levens, Eric D; Cunningham, Alexandra F; Macones, George A; Carson, Kenneth R; Beltsos, Angeline N; Odem, Randall R

    2010-09-01

    To gain a better understanding of factors influencing clinicians' embryo transfer practices. Cross-sectional survey. Web-based survey conducted in December 2008 of individuals practicing IVF in centers registered with the Society for Assisted Reproductive Technology (SART). None. None. Prevalence of clinicians reporting following embryo transfer guidelines recommended by the American Society for Reproductive Medicine (ASRM), prevalence among these clinicians to deviate from ASRM guidelines in commonly encountered clinical scenarios, and practice patterns related to single embryo transfer. Six percent of respondents reported following their own, independent guidelines for the number of embryos to transfer after IVF. Of the 94% of respondents who reported routinely following ASRM embryo transfer guidelines, 52% would deviate from these guidelines for patient request, 51% for cycles involving the transfer of frozen embryos, and 70% for patients with previously failed IVF cycles. All respondents reported routinely discussing the risks of multiple gestations associated with standard embryo transfer practices, whereas only 34% reported routinely discussing single embryo transfer with all patients. Although the majority of clinicians responding to our survey reported following ASRM embryo transfer guidelines, at least half would deviate from these guidelines in a number of different situations. Copyright (c) 2010 American Society for Reproductive Medicine. All rights reserved.

  18. Extracellular matrix synthesis in blastula and gastrula stages of normal and hybrid frog embryos

    International Nuclear Information System (INIS)

    Johnson, K.E.

    1978-01-01

    Pulse-chase labelling experiments and light- and electron-microscopic autoradiography were used to examine the site of synthesis, mode of transport, and sites of deposition of fucose-, glucose- and mannose-labelled materials in different developmental stages of normal developing Rana pipiens embryos and interspecific hybrid embryos formed by fertilizing the eggs of R. pipiens with the sperm of R. catesbeiana. In both normal and hybrid embryos, after a 15-30 min pulse, grains are closely associated with juxtanuclear and cytoplasmic collections of membrane-bound vescicles which resemble the Golgi apparatus. In normal embryos following a 15-30 min pulse and a 60-min chase, grains are largely cleared from the cytoplasmic vescicles and deposited in the extra-cellular spaces or along cell surfaces. In contrast, arrested hybrid embryos given a 15-30 min pulse and a 60-min chase show a marked accumulation of grains over cytoplasmic structures such as the Golgi apparatus and vescicular elements in the cell cortex. Certain interesting features of regional variation in synthetic activity in developing normal embryos are also described. (author)

  19. [Association of fertilization strategy and embryo transfer time with the incidence of ectopic pregnancy].

    Science.gov (United States)

    Li, Ming-zhao; Zhao, Wan-qiu; Ren, An-qi; Shi, Juan-zi

    2015-10-01

    To investigate the correlation of the fertilization strategy and embryo transfer (ET) time with the incidence of ectopic pregnancy. We selected 3,331 fresh and 2,706 frozen-thawed ET cycles for the patients undergoing in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). The fresh transfers included 2 546 IVF-ET and 785 ICSI-ET cycles and 2,220 day-3 embryo and 1,111 day-5 blastocyst transfers, while the frozen-thawed transfers included 2,080 IVF-ET and 626 ICSI-ET cycles and 741 day-3 embryo and 1 965 day-5 or -6 blastocyst transfers. We compared the incidence rate of ectopic pregnancy associated with different fertilization strategies and ET time. The incidence rate of ectopic pregnancy was 1. 41% (36/2 546) in the IVF-ET cycles and 3.44% (27/785) in the ICSI-ET cycles of the fresh transfers, significantly lower in the IVF-ET than in the ICSI-ET cycles (P 0.05). The IVF-ET and ICSI-ET cycles included 2,220 fresh day-3 (F-D3) embryos, 1,111 F-D5 blastocysts, 741 frozen-thawed day-3 (T-D3) embryos, and 1,965 T-D5/6 blastocysts. The incidence rate of ectopic pregnancy was 1.71% (n = 38) in the F-D3, 2.25% (n = 25) in the F-D5, 1.35% (n = 10) in the T-D3, and 0.81% (n = 16) in the T-D5/6 group, respectively, significantly lower in the T-D5/6 than in the other three groups (P ectopic pregnancy is associated with fertilization strategies, which is significantly lower in frozen-thawed than in fresh embryo transfers.

  20. Nuclear and cellular expression data from the whole 16-cell stage Arabidopsis thaliana embryo and a cell type-specific expression atlas of the early Arabidopsis embryo

    NARCIS (Netherlands)

    Palovaara, J.P.J.

    2017-01-01

    SuperSeries contain expression data from the nuclei of cell types involved in patterning events, with focus on root apical stem cell formation, at 16-cell stage, early globular stage and late globular stage in the early Arabidopsis embryo (atlas). Expression data comparing nuclear and cellular RNA

  1. Transferring embryos with indeterminate PGD results: the ethical implications

    OpenAIRE

    Insogna, Iris G.; Ginsburg, Elizabeth

    2016-01-01

    Background In vitro fertilization (IVF) and pre-implantation genetic diagnosis (PGD) are becoming increasingly common techniques to select embryos that are unaffected by a known genetic disorder. Though IVF-PGD has high success rates, 7.5?% of blastocysts have inconclusive results after testing. A recent case involving a known BRCA-1 carrier was brought before our Assisted Reproductive Technology Ethics Committee in order to gain a better appreciation for the ethical implications surrounding ...

  2. Effect of hyaluronic acid-enriched transfer medium on frozen-thawed embryo transfer outcomes.

    Science.gov (United States)

    Fu, Wei; Yu, Min; Zhang, Xiao-Jin

    2018-04-01

    To determine if hyaluronic acid-enriched transfer medium (HETM) affects the implantation rate (IR) and clinical pregnancy rate (PR) in women undergoing frozen-thawed embryo transfer (FET). The records of women who underwent FET from May 2014 to October 2014 were retrospectively reviewed. Outcome measures were IR and PR. In all 1721 cycles of 1632 patients were included in this study. HETM was used for 347 cycles of 342 patients, and standard medium for 1374 cycles of 1290 patients. Overall, FET outcomes were similar between the groups. For patients undergoing their first FET attempt, the IR (24.3% vs 31.6%, P = 0.042) and clinical PR (34.3% vs 50.1%, P = 0.004) were lower in the HETM group. For patients undergoing their second FET attempt, pregnancy outcomes were similar between the groups. For patients undergoing their third or more FET attempt, HETM was associated with a higher IR (33.3% vs 16.4%, P Gynecology.

  3. Acupuncture on the day of embryo transfer: a randomized controlled trial of 635 patients

    DEFF Research Database (Denmark)

    Andersen, Dorthe; Løssl, Kristine; Nyboe Andersen, Anders

    2010-01-01

    This prospective, randomized, controlled and double-blinded trial studied whether acupuncture in relation to embryo transfer could increase the ongoing pregnancy rates and live birth rates in women undergoing assisted reproductive therapy. A total of 635 patients undergoing IVF or intracytoplasmic...... sperm injection (ICSI) were included. In 314 patients, embryo transfer was accompanied by acupuncture according to the principles of traditional Chinese medicine. In the control group, 321 patients received placebo acupuncture using a validated placebo needle. In the acupuncture group and the placebo...

  4. In vitro development of canine somatic cell nuclear transfer embryos in different culture media.

    Science.gov (United States)

    Kim, Dong-Hoon; No, Jin-Gu; Choi, Mi-Kyung; Yeom, Dong-Hyeon; Kim, Dong-Kyo; Yang, Byoung-Chul; Yoo, Jae Gyu; Kim, Min Kyu; Kim, Hong-Tea

    2015-01-01

    The objective of the present study was to investigate the effects of three different culture media on the development of canine somatic cell nuclear transfer (SCNT) embryos. Canine cloned embryos were cultured in modified synthetic oviductal fluid (mSOF), porcine zygote medium-3 (PZM-3), or G1/G2 sequential media. Our results showed that the G1/G2 media yielded significantly higher morula and blastocyst development in canine SCNT embryos (26.1% and 7.8%, respectively) compared to PZM-3 (8.5% and 0%or mSOF (2.3% and 0%) media. In conclusion, this study suggests that blastocysts can be produced more efficiently using G1/G2 media to culture canine SCNT embryos.

  5. Noninvasive metabolomic profiling as an adjunct to morphology for noninvasive embryo assessment in women undergoing single embryo transfer

    NARCIS (Netherlands)

    Seli, E.; Vergouw, C.G.; Morita, H.; Botros, L.; Roos, P.; Lambalk, C.B.; Yamashita, N.; Kato, O.; Sakkas, D.

    2010-01-01

    Objective: To determine whether metabolomic profiling of spent embryo culture media correlates with reproductive potential of human embryos. Design: Retrospective study. Setting: Academic and a private assisted reproductive technology (ART) programs. Patient(s): Women undergoing single embryo

  6. Nerve growth factor regulates axial rotation during early stages of chick embryo development

    Science.gov (United States)

    Manca, Annalisa; Capsoni, Simona; Di Luzio, Anna; Vignone, Domenico; Malerba, Francesca; Paoletti, Francesca; Brandi, Rossella; Arisi, Ivan; Cattaneo, Antonino; Levi-Montalcini, Rita

    2012-01-01

    Nerve growth factor (NGF) was discovered because of its neurotrophic actions on sympathetic and sensory neurons in the developing chicken embryo. NGF was subsequently found to influence and regulate the function of many neuronal and non neuronal cells in adult organisms. Little is known, however, about the possible actions of NGF during early embryonic stages. However, mRNAs encoding for NGF and its receptors TrkA and p75NTR are expressed at very early stages of avian embryo development, before the nervous system is formed. The question, therefore, arises as to what might be the functions of NGF in early chicken embryo development, before its well-established actions on the developing sympathetic and sensory neurons. To investigate possible roles of NGF in the earliest stages of development, stage HH 11–12 chicken embryos were injected with an anti-NGF antibody (mAb αD11) that binds mature NGF with high affinity. Treatment with anti-NGF, but not with a control antibody, led to a dose-dependent inversion of the direction of axial rotation. This effect of altered rotation after anti NGF injection was associated with an increased cell death in somites. Concurrently, a microarray mRNA expression analysis revealed that NGF neutralization affects the expression of genes linked to the regulation of development or cell proliferation. These results reveal a role for NGF in early chicken embryo development and, in particular, in the regulation of somite survival and axial rotation, a crucial developmental process linked to left–right asymmetry specification. PMID:22308471

  7. Syntheses of nucleic acid and protein in somatic embryos of Fritillaria ussuriensis maxim in different development stages

    International Nuclear Information System (INIS)

    Wang Shuyu; Tang Wei; Wang Hui

    1993-09-01

    After developing a procedure for somatic embryogenesis in Fritillaria ussuriensis, dynamics on the syntheses of DNA, RNA, and protein during globular, heart-shaped, torpedo-shaped, cotyledonary, and mature somatic embryo stages was demonstrated by both autoradiography and scintillation counting. The rates of syntheses of DNA, RNA, and protein gradually increase between the globular and cotyledonary somatic embryos stages. DNA, RNA, and protein synthesis rates are in peak at the cotyledonary later stage, precotyledonary stage, and cotyledonary stage, respectively. It appears that more DNA, RNA, and protein are synthesized in the cotyledonary somatic embryo stage than in other stages. All these results indicate that an increased syntheses of DNA, RNA, and protein is associated with the differentiation of embryogenic cells and organogenesis in somatic embryos

  8. Volatiles identified from five stages of embryo development separated from a heterogeneous suspension culture of Daucus carota.

    Science.gov (United States)

    Kennedy, A H; Chamberlain, D; Wilson, G; Ryan, M F

    1991-11-01

    Five stages of embryo development were fractionated from a mature culture of Daucus carota (Gelbe Rheinsche), using a series of metal sieves. The composition of the population of embryos in each fraction was determined quantitatively from microscopic investigations. Volatiles from samples of tissue from six stages of development were trapped on activated charcoal cartridges. These volatiles, some of which may play a significant role in the interaction of the plant with the carrot root fly (Psila rosae), were analysed using gas chromatography/mass spectroscopy. The resulting chromatograms are arranged in order of embryo development. The progressive elaboration of the volatile profile reflects the increased biosynthetic capacity of the developing embryo.

  9. Evidence of Placental Autophagy during Early Pregnancy after Transfer of In Vitro Produced (IVP Sheep Embryos.

    Directory of Open Access Journals (Sweden)

    Paola Toschi

    Full Text Available Pregnancies obtained by Assisted Reproductive Technologies (ART are associated with limited maternal nutrient uptake. Our previous studies shown that in vitro culture of sheep embryos is associated with vascularization defects in their placentae and consequent reduction of embryo growth. Autophagy is a pro-survival cellular mechanism triggered by nutrient insufficiency. Therefore, the goal of our present study was to determine if autophagy is involved in early placental development after transfer of in vitro produced (IVP embryos. To do this, placentae obtained following transfer of IVP sheep embryos were compared with placentae obtained after natural mating (control-CTR. The placentae were collected on day 20 post-fertilization and post-mating, respectively, and were analyzed using molecular (qPCR, ultrastructural and histological/immunological approaches. Our results show drastically increased autophagy in IVP placentae: high levels of expression (p<0.05 of canonical markers of cellular autophagy and a high proportion of autophagic cells (35.08%; p<0.001 were observed. We conclude that high autophagic activity in IVP placentae can be a successful temporary counterbalance to the retarded vasculogenesis and the reduction of foetal growth observed in pregnancies after transfer of IVP embryos.

  10. Assisted reproduction technique outcomes for fresh versus deferred cryopreserved day-2 embryo transfer: a retrospective matched cohort study.

    Science.gov (United States)

    Bourdon, Mathilde; Santulli, Pietro; Gayet, Vanessa; Maignien, Chloé; Marcellin, Louis; Pocate-Cheriet, Khaled; Chapron, Charles

    2017-03-01

    Ovarian stimulation could adversely affect endometrial receptivity and consequently embryo implantation. One emerging strategy is the 'freeze-all' approach. Most studies have focused on blastocyst transfers, with limited research on day-2 deferred cryopreserved embryo transfers. In this large retrospective cohort study, outcomes were compared between day-2 fresh versus deferred cryopreserved embryo transfers. After matching by age and number of previous cycles, 325 cycles were included in the fresh group and 325 in the deferred cryopreserved embryo transfers group: no significant differences were found between groups in implantation (0.20 ± 0.33 versus 0.17 ± 0.31, respectively) and ongoing pregnancy rates (21.85% versus 18.46%). Independent predictors for ongoing pregnancy after a multiple logistic regression analysis were the women's age (OR = 0.92; 95% CI 0.88 to 0.97), body mass index (OR = 0.94; 95% CI 0.89 to 0.99), the number of two pronuclei embryos (OR = 1.19; 95% CI 1.04 to 1.40) and at least one grade 1 embryo transferred (OR = 1.97; 95% CI 1.26 to 3.05). In the case of a day-2 embryo transfer, outcomes after treatment with assisted reproduction techniques are similar for fresh versus deferred cryopreserved embryo transfers when pre-transfer progesterone exposures are similar in the two groups. Copyright © 2016 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  11. Considerations on the transfer of natural and artificial radionuclides from mother to embryo

    International Nuclear Information System (INIS)

    Toader, Maria; Vasilache, R. A.; Toader, Mirela; Scridon, R.

    1998-01-01

    The transfer from mother to embryo is still a matter of high interest for the radiological protection, especially when the natural radioisotopes are involved. The present paper makes a comparison between two situations, the transfer to the embryo when the mother is subjected to recurrent, constant intake of radionuclides (the case of natural radioisotopes) and the case of prolonged, variable intakes of radionuclides (the case of 137 Cs) after Chernobyl). The embryos studied were not older than 11 weeks and were taken from two areas, Bucharest (for the natural radioisotopes 226 Ra and 210 Po and for 137 Cs and 90 Sr) and uranium mining area (only for natural radioisotopes). The monitoring of the environment factors in the uranium mining area has shown that the dietary intake of natural radionuclides had extremely low variations during the interval of study. The variation of 137 Cs dietary intake was quite important during the study and was reported in previous papers. (authors)

  12. Increasing vaginal progesterone gel supplementation after frozen-thawed embryo transfer significantly increases the delivery rate

    DEFF Research Database (Denmark)

    Alsbjerg, Birgit; Polyzos, Nikolaos P; Elbaek, Helle Olesen

    2013-01-01

    The aim of this study was to evaluate the reproductive outcome in patients receiving frozen-thawed embryo transfer before and after doubling of the vaginal progesterone gel supplementation. The study was a retrospective study performed in The Fertility Clinic, Skive Regional Hospital, Denmark. A ...

  13. [Analysis of pregnancy outcomes of polycystic ovary syndrome patients after frozen embryo transfer].

    Science.gov (United States)

    Lyu, X D; Qiao, J

    2018-01-25

    Objective: To investigate pregnancy outcomes of the patients with polycystic ovary syndrome (PCOS) after frozen embryo transfer (FET) . Methods: Data of 2 367 PCOS patients received in vitro fertilization-embryo transfer [including fresh embryo transfer (fET) and FET] from January 2009 to December 2015 in Peking University Third Hospital were evaluated retrospectively. The basal characteristics, pregnancy complications and outcomes were analyzed, then identified the relative factors followed. Results: Totally 2 367 patients received in vitro fertilization-embryo transfer: 1 106 were treated with fET, and the rest 1 261 cases were treated with FET. The incidence of gestational diabetes mellitus (GDM) was lower in FET group [4.04%(51/1 261) versus 6.15%(68/1 106)], the difference was statistically significant ( Ppregnancy complications between the two groups (all P> 0.05). fET was an independent risk factor for GDM (adjusted OR= 1.570, 95% CI: 1.075-2.294). Conclusion: Compared with fET, FET could decrease the risk of GDM and receive better neonatal outcomes in patients with PCOS.

  14. Further evidence that culture media affect perinatal outcome : findings after transfer of fresh and cryopreserved embryos

    NARCIS (Netherlands)

    Nelissen, Ewka C.; Van Montfoort, Aafke P.; Coonen, Edith; Derhaag, Josien G.; Geraedts, Joep P.; Smits, Luc J.; Land, Jolande A.; Evers, Johannes L.; Dumoulin, John C.

    We have previously shown that the medium used for culturing IVF embryos affects the birthweight of the resulting newborns. This observation with potentially far-reaching clinical consequences during later life, was made in singletons conceived during the first IVF treatment cycle after the transfer

  15. Randomized trial of harp therapy during in vitro fertilization-embryo transfer.

    Science.gov (United States)

    Murphy, Erin M; Nichols, Jennifer; Somkuti, Steve G; Sobel, Michael; Braverman, Andrea; Barmat, Larry I

    2014-04-01

    This study evaluated whether harp therapy reduces levels of stress and improves clinical outcomes in patients undergoing embryo transfer. This prospective randomized trial enrolled 181 women undergoing embryo transfer, who were randomized to harp therapy during embryo transfer or standard treatment. Patients underwent standardized psychological testing and physiologic assessment of stress. The study was conducted in a reproductive medicine practice. No statistically significant differences were found in the heart and respiratory rates, nor was there a significant difference in event-based anxiety at baseline. Harp therapy had a significantly larger decrease in state anxiety from pre- to post-embryo transfer. Clinical pregnancy was 53% versus 48% for the harp therapy and standard treatment groups, respectively. Harp therapy decreases state, or event-based, anxiety, significantly lowering state scores posttransfer and having a positive effect on acute levels of stress. There was an increased pregnancy rate, but larger sample sizes are needed to evaluate whether harp therapy has an effect on clinical outcomes.

  16. EFSA Panel on Biological Hazards (BIOHAZ); Scientific Opinion on the risk of transmission of classical scrapie via in vivo derived embryo transfer in ovine animals

    DEFF Research Database (Denmark)

    Hald, Tine; Baggesen, Dorte Lau

    . Irrespective of the embryo’s genotype, embryos derived from rams and dams carrying at least one ARR allele would significantly decrease this risk (compared to an embryo from parents of unknown genotypes). The use of homozygous ARR embryos would provide the highest level of safety regarding the risk...... of transmitting classical scrapie through embryo transfer (in vivo derived embryos). The use of heterozygous ARR embryos would ensure a higher level of safety compared to Q171/Q171 embryos. Finally, it was concluded that, providing the OIE recommendations and procedures relating to embryo transfer are adhered to...

  17. Developmental disparity between in vitro-produced and somatic cell nuclear transfer bovine days 14 and 21 embryos

    DEFF Research Database (Denmark)

    Alexopoulos, Natalie I.; Maddox-Hyttel, Poul; Tveden-Nyborg, Pernille Yde

    2008-01-01

    the application of new reproductive technologies such as somatic cell nuclear transfer (SCNT). In the present study, days 14 and 21 bovine embryos, generated by either in vitro-production (IVP) or SCNT, performed by either subzonal injection (SUZI) or handmade cloning (HMC), were compared by stereomicroscopy...... of VIM and TUBB3 was less distinct in SCNT embryos when compared with IVP embryos, indicating slower or compromised development. In conclusion, SCNT and to some degree, IVP embryos displayed a high rate of embryonic mortality before D14 and surviving embryos displayed reduced quality with respect...

  18. Adverse pregnancy outcomes after in vitro fertilization: effect of number of embryos transferred and plurality at conception.

    Science.gov (United States)

    Luke, Barbara; Stern, Judy E; Kotelchuck, Milton; Declercq, Eugene R; Hornstein, Mark D; Gopal, Daksha; Hoang, Lan; Diop, Hafsatou

    2015-07-01

    To evaluate risks for adverse pregnancy outcomes by number of embryos transferred (ET) and fetal heartbeats (FHB) in assisted reproductive technology-conceived singleton live births. Longitudinal cohort using cycles reported to the Society for Assisted Reproductive Technology Clinic Outcomes Reporting System between 2004 and 2008 among women who were treated and gave birth in Massachusetts. Not applicable. Assisted reproductive technology data on 6,073 births between 2004 and 2008 were linked to vital records and hospital data. Likelihood of ET ≥3 vs. 1-2, FHB >1 vs. 1, and risks of preterm birth (PTB, 1 were modeled with binary logistic regression using a backward-stepping algorithm, and presented as adjusted odds ratios (95% confidence intervals). None. ET ≥3, FHB >1, PTB, LBW, and SGA. Higher ET was significantly more likely with older maternal age, intracytoplasmic sperm injection, assisted hatching, cleavage-stage embryos, and thawed embryos. The likelihood of FHB >1 with ≥3 ET vs. 1-2 ET was 2.04 (1.68-2.48). Risks of PTB and LBW with FHB >1 were 1.63 (1.27-2.09) and 1.81 (1.36-2.39), respectively; the risk of SGA was not significant. Nulliparity was associated with higher risks of PTB (1.34 [1.12-1.59]), LBW (1.48 [1.20-1.83]), and SGA (2.17 [1.69-2.78]). Number of embryos transferred was strongly associated with FHBs, with twice the risk of FHB >1 with ≥3 ET vs. 1-2 ET. Increasing FHBs were associated with significantly greater risks for PTB and LBW outcomes. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  19. Incorporation of 3H-thymidine during early developmental stages of ovoviviparous and oviparous embryos in the brine

    International Nuclear Information System (INIS)

    Iwasaki, Tamiko

    1980-01-01

    DNA synthesis in the embryos during various developmental stages in the brood pouch of the Artemia female as well as the encysted eggs produced by oviparity was studied by autoradiography using 3 H-thymidine. Nuclei of the embryos through the blastula stage in the brood pouch were labelled, regardless of the existence of the thick shell surrounding the embryos. However, incorporation of 3 H-thymidine into nuclei was absent in the stages between gastrula and early nauplii which had developed in the brood pouch or were delivered from encysted dry eggs. (author)

  20. Initial stages of calcium uptake and mineral deposition in sea urchin embryos

    OpenAIRE

    Vidavsky, Netta; Addadi, Sefi; Mahamid, Julia; Shimoni, Eyal; Ben-Ezra, David; Shpigel, Muki; Weiner, Steve; Addadi, Lia

    2013-01-01

    With the onset of gastrulation, sea urchin embryos deposit a calcium carbonate endoskeleton consisting of two spicules. Sea water is the source for the mineral ions, but the specific stages of the transport and deposition pathway are not well understood. This study shows that the first-formed mineral is deposited inside intracellular micrometer-size vesicles as solid nanospheres. Surprisingly, the initial deposits are distributed widely inside the embryonic cells, including epithelial cells. ...

  1. Genome stability of bovine in vivo-conceived cleavage-stage embryos is higher compared to in vitro-produced embryos.

    Science.gov (United States)

    Tšuiko, Olga; Catteeuw, Maaike; Zamani Esteki, Masoud; Destouni, Aspasia; Bogado Pascottini, Osvaldo; Besenfelder, Urban; Havlicek, Vitezslav; Smits, Katrien; Kurg, Ants; Salumets, Andres; D'Hooghe, Thomas; Voet, Thierry; Van Soom, Ann; Robert Vermeesch, Joris

    2017-11-01

    Is the rate and nature of chromosome instability (CIN) similar between bovine in vivo-derived and in vitro-cultured cleavage-stage embryos? There is a major difference regarding chromosome stability of in vivo-derived and in vitro-cultured embryos, as CIN is significantly lower in in vivo-derived cleavage-stage embryos compared to in vitro-cultured embryos. CIN is common during in vitro embryogenesis and is associated with early embryonic loss in humans, but the stability of in vivo-conceived cleavage-stage embryos remains largely unknown. Because human in vivo preimplantation embryos are not accessible, bovine (Bos taurus) embryos were used to study CIN in vivo. Five young, healthy, cycling Holstein Friesian heifers were used to analyze single blastomeres of in vivo embryos, in vitro embryos produced by ovum pick up with ovarian stimulation (OPU-IVF), and in vitro embryos produced from in vitro matured oocytes retrieved without ovarian stimulation (IVM-IVF). Single blastomeres were isolated from embryos, whole-genome amplified and hybridized on Illumina BovineHD BeadChip arrays together with the bulk DNA from the donor cows (mothers) and the bull (father). DNA was also obtained from the parents of the bull and from the parents of the cows (paternal and maternal grandparents, respectively). Subsequently, genome-wide haplotyping and copy-number profiling was applied to investigate the genomic architecture of 171 single bovine blastomeres of 16 in vivo, 13 OPU-IVF and 13 IVM-IVF embryos. The genomic stability of single blastomeres in both of the in vitro-cultured embryo cohorts was severely compromised (P vivo. Only 18.8% of in vivo-derived embryos contained at least one blastomere with chromosomal anomalies, compared to 69.2% of OPU-IVF embryos (P vivo-developed embryos, in vitro procedures exacerbate chromosomal abnormalities during early embryo development. Hence, the present study highlights that IVF treatment compromises embryo viability and should be applied

  2. Molecular asymmetry in the 8-cell stage Xenopus tropicalis embryo described by single blastomere transcript sequencing.

    Science.gov (United States)

    De Domenico, Elena; Owens, Nick D L; Grant, Ian M; Gomes-Faria, Rosa; Gilchrist, Michael J

    2015-12-15

    Correct development of the vertebrate body plan requires the early definition of two asymmetric, perpendicular axes. The first axis is established during oocyte maturation, and the second is established by symmetry breaking shortly after fertilization. The physical processes generating the second asymmetric, or dorsal-ventral, axis are well understood, but the specific molecular determinants, presumed to be maternal gene products, are poorly characterized. Whilst enrichment of maternal mRNAs at the animal and vegetal poles in both the oocyte and the early embryo has been studied, little is known about the distribution of maternal mRNAs along either the dorsal-ventral or left-right axes during the early cleavage stages. Here we report an unbiased analysis of the distribution of maternal mRNA on all axes of the Xenopus tropicalis 8-cell stage embryo, based on sequencing of single blastomeres whose positions within the embryo are known. Analysis of pooled data from complete sets of blastomeres from four embryos has identified 908 mRNAs enriched in either the animal or vegetal blastomeres, of which 793 are not previously reported as enriched. In contrast, we find no evidence for asymmetric distribution along either the dorsal-ventral or left-right axes. We confirm that animal pole enrichment is on average distinctly lower than vegetal pole enrichment, and that considerable variation is found between reported enrichment levels in different studies. We use publicly available data to show that there is a significant association between genes with human disease annotation and enrichment at the animal pole. Mutations in the human ortholog of the most animally enriched novel gene, Slc35d1, are causative for Schneckenbecken dysplasia, and we show that a similar phenotype is produced by depletion of the orthologous protein in Xenopus embryos. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  3. Increasing The Number of Embryos Transferred from Two to Three, Does not Increase Pregnancy Rates in Good Prognosis Patients

    Directory of Open Access Journals (Sweden)

    Mahnaz Ashrafi

    2015-10-01

    Full Text Available Background: To compare the pregnancy outcomes after two embryos versus three embryos transfers (ETs in women undergoing in vitro fertilization (IVF/intracytoplasmic sperm injection (ICSI cycles. Materials and Methods: This retrospective study was performed on three hundred eighty seven women with primary infertility and with at least one fresh embryo in good quality in order to transfer at each IVF/ICSI cycle, from September 2006 to June 2010. Patients were categorized into two groups according to the number of ET as follows: ET2 and ET3 groups, indicating two and three embryos were respectively transferred. Pregnancy outcomes were compared between ET2 and ET3 groups. Chi square and student t tests were used for data analysis. Results: Clinical pregnancy and live birth rates were similar between two groups. The rates of multiple pregnancies were 27 and 45.2% in ET2 and ET3 groups, respectively. The rate of multiple pregnancies in young women was significantly increased when triple instead of double embryos were transferred. Logistic regression analysis indicated two significant prognostic variables for live birth that included number and quality of transferred embryos; it means that the chance of live birth following ICSI treatment increased 3.2-fold when the embryo with top quality (grade A was transferred, but the number of ET had an inverse relationship with live birth rate; it means that probability of live birth in women with transfer of two embryos was three times greater than those who had three ET. Conclusion: Due to the difficulty of implementation of the elective single-ET technique in some infertility centers in the world, we suggest transfer of double instead of triple embryos when at least one good quality embryo is available for transfer in women aged 39 years or younger. However, to reduce the rate of multiple pregnancies, it is recommended to consider the elective single ET strategy.

  4. Body movements during early stages of chick embryo under intermittent low oxygen environment

    Science.gov (United States)

    Moriya, Kenji; Chiba, Yuya; Shimouchi, Akito

    2017-07-01

    We have attempted to elucidate the characteristic pattern of body movements in early stages of chick embryos under intermittent low oxygen incubation environment. In order to achieve this aim, the oxygen control system that can be set arbitrary oxygen concentration was developed. We choose the 18% of O2 concentration and tried to measure the embryonic body movements. As a results, only one chick embryo in the early stages under intermittent 18% O2 environment (the cycle is 18%O2-10min and 21%O2-50min) was successfully recorded and its body movements were analyzed. The characteristic body movements, which are attributed to the instantaneous effect of low oxygen environment, compared with before and after normal O2 condition were not observed. Because the early stage embryos in which the significant organs aside from heart are not formed yet have a strong adaptation to environment changes, short hypoxic condition like a 10 min might not affect instantaneous embryonic physiological changes. Meanwhile, although the cyclic interval of the large body movements becomes short in the normal development, it became long in 18%O2 condition. This result might indicate that intermittent low oxygen condition accumulatively influenced physiological function. Further improvements of accuracy in the oxygen control system and the calculation system of body movements, and further experiments under low oxygen conditions are required in the next step.

  5. Tiger, Bengal and Domestic Cat Embryos Produced by Homospecific and Interspecific Zona-Free Nuclear Transfer.

    Science.gov (United States)

    Moro, L N; Jarazo, J; Buemo, C; Hiriart, M I; Sestelo, A; Salamone, D F

    2015-10-01

    The aim of this study was to evaluate three different cloning strategies in the domestic cat (Felis silvestris) and to use the most efficient to generate wild felid embryos by interspecific cloning (iSCNT) using Bengal (a hybrid formed by the cross of Felis silvestris and Prionailurus bengalensis) and tiger (Panthera tigris) donor cells. In experiment 1, zona-free (ZP-free) cloning resulted in higher fusion and expanded blastocyst rates with respect to zona included cloning techniques that involved fusion or injection of the donor cell. In experiment 2, ZP-free iSCNT and embryo aggregation (2X) were assessed. Division velocity and blastocyst rates were increased by embryo aggregation in the three species. Despite fewer tiger embryos than Bengal and cat embryos reached the blastocyst stage, Tiger 2X group increased the percentage of blastocysts with respect to Tiger 1X group (3.2% vs 12.1%, respectively). Moreover, blastocyst cell number was almost duplicated in aggregated embryos with respect to non-aggregated ones within Bengal and tiger groups (278.3 ± 61.9 vs 516.8 ± 103.6 for Bengal 1X and Bengal 2X groups, respectively; 41 vs 220 ± 60 for Tiger 1X and Tiger 2X groups, respectively). OCT4 analysis also revealed that tiger blastocysts had higher proportion of OCT4-positive cells with respect to Bengal blastocysts and cat intracytoplasmic sperm injection blastocysts. In conclusion, ZP-free cloning has improved the quality of cat embryos with respect to the other cloning techniques evaluated and was successfully applied in iSCNT complemented with embryo aggregation. © 2015 Blackwell Verlag GmbH.

  6. Effect of Adding Human Chorionic Gonadotropin to The Endometrial Preparation Protocol in Frozen Embryo Transfer Cycles

    Directory of Open Access Journals (Sweden)

    Maryam Eftekhar

    2012-01-01

    Full Text Available Background: Human chorionic gonadotropin (HCG, one of the initial embryonic signals, isprobably a major regulator of the embryo-endometrial relationship. This study aims to assess theadvantage of HCG supplementation during the secretory phase of hormonally prepared cycles forthe transfer of cryopreserved-thawed embryos.Materials and Methods: This study was a randomized clinical trial. Infertile women who werecandidates for frozen-thawed embryo transfers entered the study and were divided into two groups,HCG and control. The endometrial preparation method was similar in both groups: all women receivedestradiol valerate (6 mg po per day from the second day of the menstrual cycle and progesteronein oil (100 mg intramuscular (I.M. when the endometrial thickness reached 8 mm. Estradiol andprogesterone were continued until the tenth week of gestation. In the HCG group, patients received anHCG 5000 IU injection on the first day of progesterone administration and the day of embryo transfer.Results: In this study, 130 couples participated: 65 in the HCG group and 65 in the control group.There was no statistically significant difference between groups regarding basic characteristics.Implantation rate, chemical pregnancy, clinical pregnancy, ongoing pregnancy, and abortion rateswere similar in both groups.Conclusion: Although HCG has some advantages in assisted reproductive technology (ARTcycles, our study did not show any benefit of HCG supplementation during the secretory phase offrozen cycles (Registration Number: IRCT201107266420N4.

  7. Embryo splitting

    Directory of Open Access Journals (Sweden)

    Karl Illmensee

    2010-04-01

    Full Text Available Mammalian embryo splitting has successfully been established in farm animals. Embryo splitting is safely and efficiently used for assisted reproduction in several livestock species. In the mouse, efficient embryo splitting as well as single blastomere cloning have been developed in this animal system. In nonhuman primates embryo splitting has resulted in several pregnancies. Human embryo splitting has been reported recently. Microsurgical embryo splitting under Institutional Review Board approval has been carried out to determine its efficiency for blastocyst development. Embryo splitting at the 6–8 cell stage provided a much higher developmental efficiency compared to splitting at the 2–5 cell stage. Embryo splitting may be advantageous for providing additional embryos to be cryopreserved and for patients with low response to hormonal stimulation in assisted reproduction programs. Social and ethical issues concerning embryo splitting are included regarding ethics committee guidelines. Prognostic perspectives are presented for human embryo splitting in reproductive medicine.

  8. Superovulation and embryo recovery in Boer goats treated with ...

    African Journals Online (AJOL)

    Lehloenya

    These pregnancy rates obtained were also indicative of an acceptable procedure of embryo transfer used in this trial. The differences obtained could also be attributed to the stage of development of the embryos, as in this trial blastocysts were used. In most trials transferable embryos (morulae and blastocysts) were used.

  9. Scriptaid Treatment Decreases DNA Methyltransferase 1 Expression by Induction of MicroRNA-152 Expression in Porcine Somatic Cell Nuclear Transfer Embryos.

    Directory of Open Access Journals (Sweden)

    Shuang Liang

    Full Text Available Abnormal epigenetic reprogramming of donor nuclei after somatic cell nuclear transfer (SCNT is thought to be the main cause of low cloning efficiencies. A growing body of evidence has demonstrated a positive role of Scriptaid, a histone deacetylase inhibitor (HDACi that belongs to an existing class of hydroxamic acid-containing HDACis, on the development competence of cloned embryos in many species. The present study investigated the effects of Scriptaid on the development of porcine SCNT embryos in vitro and its mechanism. Treatment with 300 or 500 nM Scriptaid for 20 h after activation significantly increased the percentage of SCNT embryos that developed to the blastocyst stage and the total number of cells per blastocyst and significantly decreased the percentage of apoptotic cells in blastocysts. Scriptaid treatment significantly increased the level of histone H3 acetylated at K9 and the conversion of 5-methylcytosine into 5-hydroxymethylcytosine and significantly decreased the level of histone H3 trimethylated at K9 at the pronuclear stage. As a potential mechanism for the DNA methylation changes, our results showed that the expression of DNA methyltransferase 1 was frequently down-regulated in Scriptaid-treated embryos in comparison with untreated embryos and was inversely correlated to endogenous microRNA-152 (miR-152. Taken together, these findings illustrated a crucial functional crosstalk between miR-152 and DNMT1. Meanwhile, mRNA and protein levels of POU5F1 and CDX2 were increased in Scriptaid-treated embryos. mRNA levels of Caspase3, and Bax were significantly decreased and that of Bcl-xL was significantly increased in Scriptaid-treated embryos. In conclusion, these observations would contribute to uncover the nuclear reprogramming mechanisms underlying the effects of Scriptaid on the improvement of porcine SCNT embryos.

  10. Ectopic pregnancy following in vitro fertilization with embryo transfer: A single-center experience during 15 years.

    Science.gov (United States)

    Cheng, Ling-Yun; Lin, Pin-Yao; Huang, Fu-Jen; Kung, Fu-Tsai; Chiang, Hsin-Ju; Lin, Yu-Ju; Lan, Kuo-Chung

    2015-10-01

    Ectopic pregnancy is an obstetrical disease that is potentially associated with maternal death in the first trimester. It is one of the well-known complications following in vitro fertilization (IVF) with embryo transfer (ET). The incidence of ectopic pregnancy is estimated to be 2.1-8.6% of clinical pregnancy after IVF-ET, which is higher than natural conceptions (incidence rate 2%). This study aimed to re-evaluate the ectopic pregnancy rate in patients undergoing IVF-ET and to investigate the effects of embryo stage and frozen-thawed blastocyst transfer and ET during full bladder distention on ectopic pregnancy rate. This retrospective study reviewed women who achieved a clinical pregnancy after IVF-ET at the Department of Obstetrics and Gynecology, Kaohsiung Chang Gung Memorial Hospital between 1999 and 2013. We compared ectopic pregnancy rate following Day 3 ET with Day 5 ET, and after fresh ET with thawed ET. Besides, multivariate analysis was used to clarify the factors affecting ectopic pregnancy after IVF-ET. Of the total 1213 clinical pregnancies after fresh ET, 18 (1.5%) were verified as ectopic, which is similar to the rate following natural conception. The ectopic pregnancy rates were similar for Day 3 (1.2%) and Day 5 (1.7%) ETs. The incidence of ectopic pregnancy in thawed ET cycles (0.6%) was not significantly reduced than fresh ET cycles (1.5%). Tubal ET (TET) and ET under full bladder distention had a significant effect on ectopic pregnancy. Thawed ET was not associated with a lower incidence of ectopic pregnancy than fresh ET, and embryo stage did not affect the rate of ectopic pregnancy. In addition, TET and ET under conditions of full bladder distention may increase the ectopic pregnancy rate. Copyright © 2015. Published by Elsevier B.V.

  11. The impact of consumer affordability on access to assisted reproductive technologies and embryo transfer practices: an international analysis.

    Science.gov (United States)

    Chambers, Georgina M; Hoang, Van Phuong; Sullivan, Elizabeth A; Chapman, Michael G; Ishihara, Osamu; Zegers-Hochschild, Fernando; Nygren, Karl G; Adamson, G David

    2014-01-01

    To systematically quantify the impact of consumer cost on assisted reproduction technology (ART) utilization and numbers of embryos transferred. Ordinary least squared (OLS) regression models were constructed to measure the independent impact of ART affordability-measured as consumer cost relative to average disposable income-on ART utilization and embryo transfer practices. Not applicable. Women undergoing ART treatment. None. OLS regression coefficient for ART affordability, which estimates the independent effect of consumer cost relative to income on utilization and number of embryos transferred. ART affordability was independently and positively associated with ART utilization with a mean OLS coefficient of 0.032. This indicates that, on average, a decrease in the cost of a cycle of 1 percentage point of disposable income predicts a 3.2% increase in utilization. ART affordability was independently and negatively associated with the number of embryos transferred, indicating that a decrease in the cost of a cycle of 10 percentage points of disposable income predicts a 5.1% increase in single-embryo transfer cycles. The relative cost that consumers pay for ART treatment predicts the level of access and number of embryos transferred. Policies that affect ART funding should be informed by these findings to ensure equitable access to treatment and clinically responsible embryo transfer practices. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  12. AFSC/RACE/SAP/Foy: Effects of ocean acidification on embryo stages of Tanner crab: Kodiak Island, Alaska.

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — To study the effects of ocean acidification we examined the effects of ocean acidification on the embryo stages of the economically important southern Tanner crab,...

  13. DNA methylation in porcine preimplantation embryos developed in-vivo or produced by in-vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer

    DEFF Research Database (Denmark)

    Deshmukh, Rahul Shahaji; Østrup, Olga; Østrup, Esben

    2011-01-01

    ), in vitro fertilized (IVF), somatic cell nuclear transfer (SCNT) and parthenogenetically activated (PA) embryos were evaluated for DNA methylation quantification at different developmental stages. Fertilized (IV and IVF) one-cell stages lacked a substantial active demethylation of the paternal genome......DNA demethylation and remethylation are crucial for reprogramming of the differentiated parental/somatic genome in the recipient ooplasm upon somatic cell nuclear transfer. Here, we analyzed the DNA methylation dynamics during porcine preimplantation development. Porcine in vivo developed (IV....... Embryos produced under in vitro conditions had higher levels of DNA methylation than IV. A lineage-specific DNA methylation (hypermethylation of the inner cell mass and hypomethylation of the trophectoderm) was observed in porcine IV late blastocysts, but was absent in PA- and SCNT-derived blastocysts...

  14. DNA methylation in porcine preimplantation embryos developed in vivo and produced by in vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer

    DEFF Research Database (Denmark)

    Deshmukh, Rahul Shahaji; Østrup, Olga; Østrup, Esben

    2011-01-01

    ), in vitro fertilized (IVF), somatic cell nuclear transfer (SCNT) and parthenogenetically activated (PA) embryos were evaluated for DNA methylation quantification at different developmental stages. Fertilized (IV and IVF) one-cell stages lacked a substantial active demethylation of the paternal genome......DNA demethylation and remethylation are crucial for reprogramming of the differentiated parental/somatic genome in the recipient ooplasm upon somatic cell nuclear transfer. Here, we analyzed the DNA methylation dynamics during porcine preimplantation development. Porcine in vivo developed (IV....... Embryos produced under in vitro conditions had higher levels of DNA methylation than IV. A lineage-specific DNA methylation (hypermethylation of the inner cell mass and hypomethylation of the trophectoderm) was observed in porcine IV late blastocysts, but was absent in PA- and SCNT-derived blastocysts...

  15. Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos.

    Science.gov (United States)

    Huang, Yongye; Tang, Xiaochun; Xie, Wanhua; Zhou, Yan; Li, Dong; Zhou, Yang; Zhu, Jianguo; Yuan, Ting; Lai, Liangxue; Pang, Daxin; Ouyang, Hongsheng

    2011-07-29

    The reprogramming of differentiated cells into a totipotent embryonic state through somatic cell nuclear transfer (SCNT) is still an inefficient process. Previous studies revealed that the generation of induced pluripotent stem (iPS) cells from mouse and human fibroblasts could be significantly enhanced with vitamin C treatment. Here, we investigated the effects of vitamin C, to our knowledge for the first time, on the in vitro and in vivo development of porcine SCNT embryos. The rate of blastocyst development in SCNT embryos treated with 50μg/mL vitamin C 15h after activation (36.0%) was significantly higher than that of untreated SCNT embryos (11.5%). The enhanced in vitro development rate of vitamin C-treated embryos was associated with an increased acetylation level of histone H4 lysine 5 and higher Oct4, Sox2 and Klf4 expression levels in blastocysts, as determined by real-time PCR. In addition, treatment with vitamin C resulted in an increased pregnancy rate in pigs. These findings suggest that treatment with vitamin C is beneficial for enhancement of the in vitro and in vivo development of porcine SCNT embryos. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. The probability of pregnancy after embryo transfer is affected by the age of the patient, cause of infertility, number of embryos transferred and the average morphology score, as revealed by multiple logistic regression analysis

    NARCIS (Netherlands)

    Roseboom, T. J.; Vermeiden, J. P.; Schoute, E.; Lens, J. W.; Schats, R.

    1995-01-01

    Because the process of conception is affected by many variables, a multiple logistic regression analysis was performed to assess (i) the impact and relative weight of both patient and embryo variables and (ii) their possible effects on the probability of a vital pregnancy after embryo transfer. A

  17. Mosaic embryo transfer after oocyte in vitro maturation in combination with non-invasive prenatal testing (NIPT)-first report of a euploid live birth.

    Science.gov (United States)

    Inoue, Naomi; Lopez, Rosmary; Delgado, Andrea; Nuñez, Denisse; Portella, Jimmy; Noriega-Hoces, Luis; Guzmán, Luis

    2017-09-01

    The purpose of this study is to describe a healthy life birth after a mosaic embryo transfer in oocyte in vitro maturation (IVM). Patient received minimal stimulation, starting on day 3 after menstrual period. No hCG trigger was administered. Oocyte retrieval was performed and oocytes were matured for 30 h. After denuding, mature oocytes were inseminated by ICSI. Embryos were cultured until blastocyst stage and biopsied. One euploid embryo after array comprehensive genome hybridization (aCGH) was diagnostic. However, the next-generation sequencing (NGS) re-analysis showed that embryo was a mosaic for chromosome 13 and 21. Nevertheless, pregnancy ultrasound scans and non-invasive prenatal test (NIPT-Verifi-Illumina) indicated a normal fetus development. Finally, a healthy baby was born after 38 weeks. Its weight was 4480 g, head circumference 36 cm, and total length of 51 cm. To confirm that the baby was chromosomically normal, an NGS test was performed in buccal cells, a normal profile was obtained. Our finding confirmed that mosaic embryo transfer would bring a healthy offspring.

  18. Large scale in vivo risk assessment of bovine viral diarrhea virus (BVDV) transmission through transfer of bovine embryos produced via somatic cell nuclear transfer (SCNT).

    Science.gov (United States)

    Gregg, K; Gosch, G; Guerra, T; Chen, S H; Xiang, T; Broek, D; Bruner, B; Polejaeva, I

    2010-10-15

    The objective was to use the bovine viral diarrhea virus (BVDV) as a model to assess the risk of infectious disease transmission in the system of in vitro embryo production and transfer via somatic cell nuclear transfer (SCNT) technology. The risks of BVDV transmission in the SCNT embryo production were previously evaluated. In that in vitro study, following standard operating procedures (SOP), including pre-nuclear transfer donor cell testing, oocyte decontamination and virus-free cell and embryo culture conditions, SCNT embryos produced were free of detectable viral RNA. The current study focused on the evaluation of the potential risk of disease transmission from SCNT embryos to recipients, and the risk of producing persistently infected animals via SCNT embryo transfer. Blood samples were collected from 553 recipients of SCNT embryos and 438 cloned calves and tested for the presence of BVDV viral RNA via a sensitive real time PCR method. All samples tested were negative. These results, in conjunction with the previous in vitro study, confirmed that the established SCNT embryo production and transfer system is safe and presents no detectable risk of disease transmission. Copyright © 2010 Elsevier Inc. All rights reserved.

  19. Vitrification of pronuclear-stage mouse embryos on solid surface (SSV) versus in cryotube: comparison of the effect of equilibration time and different sugars in the vitrification solution.

    Science.gov (United States)

    Bagis, H; Sagirkaya, H; Mercan, H Odaman; Dinnyès, A

    2004-02-01

    The cryopreservation of pronuclear-stage embryos has particular importance in transgenic technology and human assisted reproductive technology (ART). The objective of this study was to improve the efficiency of cryopreservation of pronuclear-stage mouse embryos. Two vitrification methods (solid surface vitrification (SSV) vs. vitrification in cryotube) have been compared with special emphasis on the effect of the exposure of the embryos to the solutions for various times and the sugar content (trehalose, sucrose, or raffinose) of the vitrification solutions. Pronuclear-stage embryos were either exposed to 1 M dimethyl sulfoxide (DMSO) + 1 M propylene-glycol (PG) solution for 2, 5, 10, or 15 min or not exposed to this "equilibration" solution. The vitrification solutions consisted of 2.75 M DMSO and 2.75 M PG in M2 medium supplemented with 1 M trehalose (DPT), 1 M sucrose (DPS), or 1 M raffinose (DPR). In the cryotube method, groups of 15-25 embryos were transferred into a 1.8 ml cryotube containing 30 microl of DPT, DPS, or DPR. After 30 sec, the cryotubes were directly plunged into liquid nitrogen (LN(2)) and stored for 1 day to 1 month. Vitrified samples were warmed by immersing the cryotubes in a 40 degrees C water bath and then immediately diluted with 300 microl of 0.3 M trehalose, sucrose, or raffinose in M2. In the SSV method, after equilibration 15-20 embryos were exposed to DPT, DPS, or DPR solutions for around 20 sec before being dropped in 2-microl drops onto a pre-cooled (-150 to -180 degrees C) metal surface. Vitrified droplets were stored in cryovials in LN(2). Warming was performed by transferring the vitrified droplets into 0.3 M solutions of trehalose, sucrose, or raffinose at 37 degrees C, respectively. Results showed that both SSV and cryotube vitrification methods can result in high rates of in vitro blastocyst development (up to 58.3 and 68.5% with DPR, respectively), not statistically different from that of the controls (58.3 and 64.4%). Even

  20. Cloning endangered felids using heterospecific donor oocytes and interspecies embryo transfer.

    Science.gov (United States)

    Gómez, Martha C; Pope, C Earle; Ricks, David M; Lyons, Justine; Dumas, Cherie; Dresser, Betsy L

    2009-01-01

    Somatic cell nuclear transfer (SCNT) offers the possibility of preserving endangered species. It is one of the few technologies that avoids the loss of genetic variation and provides the prospect of species continuance, rather than extinction. Nonetheless, there has been a debate over the use of SCNT for preserving endangered species because of abnormal nuclear reprogramming, low efficiency and the involvement of extra mitochondrial DNA (mtDNA) of a different species in live offspring produced by interspecies SCNT. Despite these limitations, live endangered cloned animals have been produced. In the present paper, we describe recent research on the production of cloned embryos derived by fusion of wild felid fibroblast cells with heterospecific domestic cat cytoplasts and their viability after transfer into domestic cat recipients. In addition, we discuss epigenetic events that take place in donor cells and felid cloned embryos and mtDNA inheritance in wild felid clones and their offspring.

  1. Acupuncture on the day of embryo transfer: a randomized controlled trial of 635 patients

    DEFF Research Database (Denmark)

    Andersen, Dorota; Løssl, Kristine; Nyboe Andersen, Anders

    2010-01-01

    This prospective, randomized, controlled and double-blinded trial studied whether acupuncture in relation to embryo transfer could increase the ongoing pregnancy rates and live birth rates in women undergoing assisted reproductive therapy. A total of 635 patients undergoing IVF or intracytoplasmic...... sperm injection (ICSI) were included. In 314 patients, embryo transfer was accompanied by acupuncture according to the principles of traditional Chinese medicine. In the control group, 321 patients received placebo acupuncture using a validated placebo needle. In the acupuncture group and the placebo...... group, the ongoing pregnancy rates were 27% (95% CI 22-32) and 32% (95% CI 27-37), respectively. Live birth rates were 25% (95% CI 20-30) in the acupuncture group and 30% (95% CI 25-30) in the placebo group. The differences were not statistically significant. These results suggest that acupuncture...

  2. NATIONAL PROGRAM FOR IN VITRO FERTILIZATION AND EMBRYO TRANSFER IN ROMANIA: ETHICAL, LEGAL, AND SOCIAL CHALLENGES

    Directory of Open Access Journals (Sweden)

    Gabriela SIMIONESCU

    2017-05-01

    Full Text Available This review summarizes aspects regarding the national program for in vitro fertilization and embryo transfer in Romania, emphasizing on the ethical, legal and social challenges associated with assisted reproduction technologies. Romania is one of the few countries from the European Union that does not have a specific law for human assisted reproduction, but infertile couples in Romania may benefit from the national program for in vitro fertilization and embryo transfer although, unfortunately, the allocated public funds are not in line with the demand. There are a series of inclusion criteria when applying for the program and unlike other countries, only one in vitro fertilization (IVF procedure may be publicly funded. Despite the legal, ethical and social challenges, this program, however, represents an extremely important step in aligning our country with the standards of other developed countries.

  3. Establishment of pregnancy after the transfer of nuclear transfer embryos produced from the fusion of argali (Ovis ammon) nuclei into domestic sheep (Ovis aries) enucleated oocytes.

    Science.gov (United States)

    White, K L; Bunch, T D; Mitalipov, S; Reed, W A

    1999-01-01

    by exposure to ionomycin (5 microM, 4 minutes) followed by incubation in 6-dimethylaminopurine (0.2 mM, 4 hours) and cultured in microdrops of CR1aa medium. From a total of 166 constructed nuclear donor cell-bovine cytoplasm NT couples, 128 (77%) successfully fused, 100 (78%) developed to 8-16 cell stage, and 2 (1.56%) developed to the blastocyst stage. The presence of argali nuclei in 8-16 cell stage embryo clones was confirmed after observation of Hoechst 33342 stained embryos under UV light and chromosome analysis of metaphase spreads from blastomeres. A total of 127 constructed nuclear donor cell-ovine cytoplasm NT couples were produced, 101 (80%) successfully fused, 81 (80% of fused) developed to the 16- to 32-cell stage. A total of 28 hybrid (argali-sheep) and 21 sheep-sheep NT embryos were transferred into 6 recipients and 4 recipients, respectively. Two of these recipients, 1 carrying argali-sheep and 1 sheep-sheep, were confirmed pregnant at 49 days by ultrasound, but both pregnancies terminated by 59 days. The results of this study demonstrate the possibility of using xenogenic oocytes to produce early-stage embryos and pregnancies from an established fibroblast cell line of an endangered species.

  4. The effects of embryo culture media on the birthweight of singletons via fresh or frozen-thawed embryo transfer: a large-scale retrospective study.

    Science.gov (United States)

    Gu, Fang; Deng, Mingfen; Gao, Jun; Wang, Zilian; Ding, Chenhui; Xu, Yanwen; Zhou, Canquan

    2016-09-19

    Embryo culture media used for IVF treatment might affect fetal growth and thus birthweight of the newborns. A retrospective study was conducted in South China using data from 2370 singleton neonates born after IVF/ICSI between 2009 and 2012. Two culture media, i.e., either Vitrolife or SAGE were used as embryo culture media during the study period. Neonates' birthweights were compared between the two embryo culture media groups. Among the 2370 singletons, 1755 cases came from fresh cleavage embryo transfer while 615 were from frozen-thawed cleavage embryo transfer. Within the fresh embryo transfer newborns, no statistical difference was observed in either birthweight (mean ± SD: 3196.0 ± 468.9 versus 3168.4 ± 462.0g, p > 0.05) or adjusted birthweight controlled for gestational age and gender (z-score mean ± SD: 0.11 ± 1.02 versus 0.11 ± 0.99 g, P > 0.05) between the Vitrolife (n = 419) and the SAGE group (n = 1336). Likewise within frozen embryo transfer neotates, no statistical difference of the birthweight (3300.6 ± 441.3 vs.3256.0 ± 466.7 g, P > 0.05) and adjusted birthweight (0.30 ± 0.99 g versus 0.29 ± 0.97 g, P > 0.05) was found between the Vitrolife (n = 202) and the SAGE group (n = 413). The sex ratio [OR1.17, 95 % CI (0.94-1.46)/OR1.1, 95 % CI (0.78-1.54)], rate of small for gestational age [OR1.14, 95 % CI (0.82-1.59)/OR1.06, 95 % CI (0.56-2.02)] and large for gestational age [OR1.07, 95 % CI (0.64-1.76)/OR0.98, 95 % CI (0.47-2.02)] in fresh and frozen-thawed subgourps are all comparable respectively between the two culture media. No group differences were found in the rate of low birthweight and macosomia. Multiple linear regression analysis demonstrated that maternal weight, gestational age, frozen-thawed embryo transfer and infant gender were significantly related to neonatal birthweight (P cultured in SAGE or Vitrolife media after fresh or frozen-thawed cleavage

  5. Current Status of Comprehensive Chromosome Screening for Elective Single-Embryo Transfer

    OpenAIRE

    Wu, Ming-Yih; Chao, Kuang-Han; Chen, Chin-Der; Chang, Li-Jung; Chen, Shee-Uan; Yang, Yu-Shih

    2014-01-01

    Most in vitro fertilization (IVF) experts and infertility patients agree that the most ideal assisted reproductive technology (ART) outcome is to have a healthy, full-term singleton born. To this end, the most reliable policy is the single-embryo transfer (SET). However, unsatisfactory results in IVF may result from plenty of factors, in which aneuploidy associated with advanced maternal age is a major hurdle. Throughout the past few years, we have got a big leap in advancement of the genetic...

  6. Transcriptional reprogramming of gene expression in bovine somatic cell chromatin transfer embryos

    Directory of Open Access Journals (Sweden)

    Page Grier P

    2009-04-01

    Full Text Available Abstract Background Successful reprogramming of a somatic genome to produce a healthy clone by somatic cells nuclear transfer (SCNT is a rare event and the mechanisms involved in this process are poorly defined. When serial or successive rounds of cloning are performed, blastocyst and full term development rates decline even further with the increasing rounds of cloning. Identifying the "cumulative errors" could reveal the epigenetic reprogramming blocks in animal cloning. Results Bovine clones from up to four generations of successive cloning were produced by chromatin transfer (CT. Using Affymetrix bovine microarrays we determined that the transcriptomes of blastocysts derived from the first and the fourth rounds of cloning (CT1 and CT4 respectively have undergone an extensive reprogramming and were more similar to blastocysts derived from in vitro fertilization (IVF than to the donor cells used for the first and the fourth rounds of chromatin transfer (DC1 and DC4 respectively. However a set of transcripts in the cloned embryos showed a misregulated pattern when compared to IVF embryos. Among the genes consistently upregulated in both CT groups compared to the IVF embryos were genes involved in regulation of cytoskeleton and cell shape. Among the genes consistently upregulated in IVF embryos compared to both CT groups were genes involved in chromatin remodelling and stress coping. Conclusion The present study provides a data set that could contribute in our understanding of epigenetic errors in somatic cell chromatin transfer. Identifying "cumulative errors" after serial cloning could reveal some of the epigenetic reprogramming blocks shedding light on the reprogramming process, important for both basic and applied research.

  7. Future aspects of micromanipualtion with embryos for

    African Journals Online (AJOL)

    Embryo micromanipulation techniques and their potential genetic impact in dairy cattle are discussed. In addition, some aspects of gene transfer are mentioned. Only the technique of splitting bovine embryos and the subsequent transfer of halfembryos has reached a stage which might make its application to cattle breeding ...

  8. Totipotency segregates between the sister blastomeres of two-cell stage mouse embryos.

    Science.gov (United States)

    Casser, E; Israel, S; Witten, A; Schulte, K; Schlatt, S; Nordhoff, V; Boiani, M

    2017-08-15

    Following fertilization in mammals, it is generally accepted that totipotent cells are exclusive to the zygote and to each of the two blastomeres originating from the first mitotic division. This model of totipotency was inferred from a minority of cases in which blastomeres produced monozygotic twins in mice. Was this due to experimental limitation or biological constraint? Here we removed experimental obstacles and achieved reliable quantification of the prevalence of dual totipotency among mouse two-cell stage blastomeres. We separated the blastomeres of 1,252 two-cell embryos, preserving 1,210 of the pairs. Two classes of monozygotic twins became apparent at the blastocyst stage: 27% formed a functional epiblast in both members (concordant), and 73% did so in only one member of the pair (discordant) - a partition that proved insensitive to oocyte quality, sperm-entry point, culture environment and pattern of cleavage. In intact two-cell embryos, the ability of sister blastomeres to generate epiblast was also skewed. Class discovery clustering of the individual blastomeres' and blastocysts' transcriptomes points to an innate origin of concordance and discordance rather than developmental acquisition. Our data place constraints on the commonly accepted idea that totipotency is allocated equally between the two-cell stage blastomeres in mice.

  9. Stage-dependent ethoxyresorufin-O-deethylase (EROD) in vivo activity in medaka (Oryzias latipes) embryos.

    Science.gov (United States)

    González-Doncel, Miguel; Carbonell, Gregoria; San Segundo, Laura; Sastre, Salvador; Beltrán, Eulalia M; Fernández-Torija, Carlos

    2015-09-01

    Using medaka (Oryzias latipes) embryos, this study aimed to quantitatively characterize the stage-dependent in vivo ethoxyresorufin-O-deethylase (EROD) as indicator of cytochrome P4501A (CYP1A) activity. Embryos were challenged for 24-h to an agonist (β-naphthoflavone [BNF], 2.5, 5, 10, and 20 μg L(-1)) or to its combination (2.5 μg L(-1)) with an antagonist (α-naphthoflavone [ANF], 25, 50, 100, and 200 μg L(-1)), initiated at four different developmental time points (1, 3, 6, and 9 d post-fertilization [dpf]). Respective induction and competitive inhibition were evaluated over fluorescent images of whole embryo (nonorgan-specific [NOS] EROD activity) and gallbladder (organ-specific [OS] EROD activity). Both flavonoids showed signs of stability in solution. Generally speaking, the mean fluorescence intensity (MFI) values for NOS EROD increased with BNF concentration and exposure challenge. BNF co-exposure with ⩾50 μg ANF L(-1) during the 1-2 and 3-4 dpf challenges lowered NOS EROD to undetectably induced levels. Significant increments in MFIs for OS-EROD were seen from exposures to ⩾2.5 μg BNF L(-1), peaking during the 6-7 dpf challenge regardless of BNF concentration. The simultaneous BNF/ANF incubation showed competitive inhibition for OS EROD activity, although levels were generally detectably induced during all challenges and at all ANF concentrations. The morphometric in vivo gallbladder analysis indicated significant dilation in the 10 dpf-old embryos co-exposed to BNF and 200 μg ANF L(-1). This quantitative approach can be used successfully at 4 dpf at the NOS-EROD or OS-EROD levels, although the NOS-EROD response was sensitive enough to induction or inhibition, even at 2 dpf. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Comparison of ectopic pregnancy risk among transfers of embryos vitrified on day 3, day 5, and day 6.

    Science.gov (United States)

    Du, Tong; Chen, Hong; Fu, Rong; Chen, Qiuju; Wang, Yun; Mol, Ben W; Kuang, Yanping; Lyu, Qifeng

    2017-07-01

    To compare ectopic pregnancy risk among transfers of embryos vitrified on day 3, day 5, and day 6. Retrospective cohort study. Academic tertiary-care medical center. A total of 10,736 pregnancies after 23,730 frozen-thawed embryo transfer (FET) cycles of in vitro fertilization/intracytoplasmic sperm injection from March 2003 to May 2015. The ectopic pregnancy rate was compared among pregnancies resulting from transfers of embryos vitrified on day 3, day 5, and day 6. Generalized estimation equation regression models were used to calculate unadjusted and adjusted odds ratios and 95% confidence intervals for the association between ectopic pregnancy and selected patient and treatment characteristics. We studied this association in both the group that achieved pregnancy and the group that underwent an FET cycle. Odds of ectopic pregnancy. The overall rate of ectopic pregnancy was 2.8% (304/10,736). Ectopic pregnancy rates after day-3, day-5, and day-6 vitrified embryo transfers were 3.1% (287/9,224), 2.0% (11/562), and 0.6% (6/950), respectively. After adjusting for confounders, the risks of ectopic pregnancy in day-3 and day-5 vitrified embryo transfers were both significantly higher than in day-6 vitrified embryo transfers. The associations were similar when we did calculations per cycle. In women undergoing FET, day-6 vitrified embryo transfer is associated with a significantly lower risk of ectopic pregnancy than both day-3 and day-5 vitrified embryo transfers. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  11. Material Transfer Agreement (MTA) | FNLCR Staging

    Science.gov (United States)

    Material Transfer Agreements are appropriate for exchange of materials into or out of the Frederick National Labfor research or testing purposes, with no collaborative research by parties involving the materials.

  12. Assisted Reproductive Technology and Newborn Size in Singletons Resulting from Fresh and Cryopreserved Embryos Transfer.

    Directory of Open Access Journals (Sweden)

    Galit Levi Dunietz

    Full Text Available The aim of this study was two-fold: to investigate the association of Assisted Reproductive Technology (ART and small newborn size, using standardized measures; and to examine within strata of fresh and cryopreserved embryos transfer, whether this association is influenced by parental infertility diagnoses. We used a population-based retrospective cohort from Michigan (2000-2009, Florida and Massachusetts (2000-2010. Our sample included 28,946 ART singletons conceived with non-donor oocytes and 4,263,846 non-ART singletons.Regression models were used to examine the association of ART and newborn size, measured as small for gestational age (SGA and birth-weight-z-score, among four mutually exclusive infertility groups: female infertility only, male infertility only, combined female and male infertility, and unexplained infertility, stratified by fresh and cryopreserved embryos transfer.We found increased SGA odds among ART singletons from fresh embryos transfer compared with non-ART singletons, with little difference by infertility source [adjusted odds-ratio for SGA among female infertility only: 1.18 (95% CI 1.10, 1.26, male infertility only: 1.20 (95% CI 1.10, 1.32, male and female infertility: 1.18 (95% CI 1.06, 1.31 and unexplained infertility: 1.24 (95% CI 1.10, 1.38]. Conversely, ART singletons, born following cryopreserved embryos transfer, had lower SGA odds compared with non-ART singletons, with mild variation by infertility source [adjusted odds-ratio for SGA among female infertility only: 0.56 (95% CI 0.45, 0.71, male infertility only: 0.64 (95% CI 0.47, 0.86, male and female infertility: 0.52 (95% CI 0.36, 0.77 and unexplained infertility: 0.71 (95% CI 0.47, 1.06]. Birth-weight-z-score was significantly lower for ART singletons born following fresh embryos transfer than non-ART singletons, regardless of infertility diagnoses.

  13. Elective single-embryo transfer: persuasive communication strategies can affect choice in a young British population.

    Science.gov (United States)

    van den Akker, O B A; Purewal, S

    2011-12-01

    This study tested the effectiveness of the framing effect and fear appeals to inform young people about the risks of multiple births and the option of selecting elective single-embryo transfer (eSET). A non-patient student sample (age (mean±SD) 23±5.5 years; n=321) were randomly allocated to one of seven groups: (1) framing effect: (1a) gain and (1b) loss frame; (2) fear appeal: (2a) high, (2b) medium and (2c) low fear; or (3) a control group: (3a) education and (3b) non-education. The primary outcome measure was the Attitudes towards Single Embryo Transfer questionnaire, before exposure to the messages (time 1) and immediately afterwards (time 2). Results revealed participants in the high fear, medium fear and gain condition demonstrated the most positive and significant differences (Ppersuasive communication techniques on a student population to promote immediate and hypothetical eSET preferences is more successful at promoting eSET than merely reporting educational content. Future research should investigate its application in a clinical population. A multiple pregnancy is a health risk to both infant and mother following IVF treatment. The aims of this study were to test the effectiveness of two persuasive communication techniques (the framing effect and fear appeals) to inform young people about the risks of multiple births and the hypothetical option of selecting elective single-embryo transfer (eSET) (i.e., only one embryo is transferred to the uterus using IVF treatment). A total of 321 non-patient student sample (mean age 23) were randomly allocated to read a message from one of seven groups: (1) framing effect: (1a) gain and (1b) loss frame; (2) fear appeal: (2a) high, (2b) medium and (2c) low fear; or (3) a control group: education (3a) and (3b) non-education. Participants completed the Attitudes towards Single Embryo Transfer questionnaire, before exposure to the messages (time 1) and immediately afterwards (time 2). Results revealed that participants

  14. Use of purified FSH and LH for embryo production, cryopreservation by conventional freezing or vitrification and transfer of embryos in dairy ewes

    Directory of Open Access Journals (Sweden)

    Giovanni Martemucci

    2010-01-01

    Full Text Available Three experiments were carried out with the aim of evaluating the efficiency of techniques of in vivo production, storageand transfer of embryos in dairy sheep. Experiment I - For embryo production, thirty-one ewes were synchronized withFGA (vaginal sponges, 40 mg, 9 d and PGF2α (ICI; 50 μg, 7th d, and subdivided into three groups corresponding to thefollowing superovulatory treatments over 3 days with purified gonadotrophic preparations: A control, FSH/LH ratio = 1(250 IU p-FSH : 250 UI p-LH; B FSH/LH ratio = 2 (250 IU p-FSH : 125 IU p-LH and daily FSH/LH ratio of 3.4 – 1.7 –0.8 in the 3 days of treatment, respectively; C FSH/LH ratio = 2 (250 IU p-FSH : 125 IU p-LH and daily FSH/LH ratioof 5.0 – 1.0 – 0.3. On the 7th day after oestrus and mating, ovarian response and embryo production were evaluated.Experiment II – Three freezing methods were evaluated based upon post-thaw embryo quality: CF conventional slowfreezing by 1.5 M ethylene glycol (EG; V-1 one-step vitrification based on exposure of the embryos to one solution (EG7.15 M + ficoll 2.5 mM; V-3 vitrification in three steps, corresponding to three solutions at increasing concentration ofglycerol (GLY and EG (GLY 1.4 M; GLY 3.4 M + EG 1.4 M; GLY 4.6 M + EG 3.4 M. V-1 and V-3 frozen embryos weredirectly plunged in liquid nitrogen. At thawing, embryo viability was evaluated on the basis of morphological features.Experiment III – For embryo transfer, a total of 26 recipient ewes were synchronized with donors. On the 7th d fromoestrus, 11 recipient ewes received fresh embryos (Group FE – control and 15 recipients received vitrified-thawedembryos (Group VTE. Each recipient received 2 embryos. Superovulatory treatment B significantly advanced the onsetof oestrus compared to the control (27.3 vs 34.7 h; P10.8. Transferable embryos in Group B (7.2 resulted similar to Group A (5.3 and significantly (Pcompared to Group C (3.2. V3-method resulted in the highest (PCF- and V1-methods

  15. [Pregnancy and obstetric outcomes of fresh embryo transfer versus frozen-thawed embryo transfer in women below 35 years of age].

    Science.gov (United States)

    Sun, Ling; Chen, Zhi-Heng; Yin, Min-Na; Deng, Yu

    2017-07-20

    To compare the obstetric and perinatal outcomes between fresh embryo transfer (ET) and frozen-thawed ET (the "freeze-all" strategy) and evaluate the benefits of the "freeze-all" embryo strategy for young patients. We reviewed a total of 2091 ET cycles performed between January, 2011 and December, 2015 in women aged 20-35 years, including 1295 fresh ET cycles and 796 frozen-thawed ET cycles. The demographic characteristics, ovarian stimulation syndrome, clinical pregnancy rates, live birth rate and the obstetric outcomes (gestational age, preterm delivery rate and mean birth weight) were compared between the two groups. The mean age of the patients receiving frozen-thawed ET cycles had a significantly younger age than those having fresh ET cycles (29.5 vs 30.2 years, P0.05), live birth rate (50.3% vs 47.0%; P>0.05), mean birth weight or gestational age between the two groups. The freeze-all policy produces similar pregnancy and obstetric outcomes with those of fresh ET. Our results support the hypothesis that the freeze-all strategy help to prevent OHSS with a good pregnancy rate.

  16. Effect of frozen/thawed embryo transfer on birthweight, macrosomia, and low birthweight rates in US singleton infants.

    Science.gov (United States)

    Litzky, Julia F; Boulet, Sheree L; Esfandiari, Navid; Zhang, Yujia; Kissin, Dmitry M; Theiler, Regan N; Marsit, Carmen J

    2017-12-29

    Singleton infants conceived using assisted reproductive technology have lower average birthweights than naturally conceived infants and are more likely to be born low birthweight (macrosomia (>4000 g) and low birthweight (macrosomia and low birthweight were evaluated using multivariable predicted marginal proportions from logistic regression models. In total, 180,184 singleton, term infants were included, with 55,898 (31.02%) having been conceived from frozen/thawed embryos. Frozen/thawed embryo transfer was associated with, on average, a 142 g increase in birthweight compared with infants born after fresh embryo transfer (P macrosomia following frozen/thawed embryo transfer was greater than that following fresh embryo transfer, but the risk of low birthweight among frozen/thawed embryo transfer infants was significantly decreased in comparison with fresh embryo transfer infants. Copyright © 2018 Elsevier Inc. All rights reserved.

  17. Transfer line tests take centre stage

    CERN Document Server

    Katarina Anthony

    2014-01-01

    Last weekend, proton beams came knocking on the LHC's door. Shooting from the SPS and into the two LHC transfer lines, the proton beams were dumped just short of entering the accelerator.   The upper plot shows the trajectory of the first TI2 beam, which reached the end of the transfer line in a single attempt after 18 months of technical stop. Below, a smoother beam trajectory in TI2 after some corrections. For the first time since Run 1, the SPS to LHC transfer lines (TI8 and TI2) transported proton beams just short of the LHC. "We tested the beam instrumentation, the devices that measure the beam intensity, transverse beam profile, position and losses, as well as the beam collimators along the transfer lines," says Reyes Alemany Fernandez, the engineer in charge of the LHC. "We were also able to spot possible bottle necks in the beam trajectory and to perform the first optics measurements." Once the beams arrived at the transfer line beam dumps...

  18. [The effect of warm needling pretreatment for endometrial receptivity of frozen embryo transfer].

    Science.gov (United States)

    Luo, Guoqun; Su, Wenwu; Ma, Wenmin; Situ, Yanqiu; Xie, Chongju; Yang, Meiqiong

    2017-08-12

    To explore the effect of warming needling pretreatment for endometrial receptivity before frozen-thawed embryo transfer (FET). Fifty-six repeatedly embryo transfer (ET) failure patients with ultrasound showing follicular phase endometrium of C type, hysteroscopy examination presenting endometritis were randomly assigned into an observation group (25 cases) and a control group (31 cases). The patients in the observation group three months before ET were treated with antibiotics in the menstrual period, warming needle (once a day) at Zhongwan (CV 12), Tianshu (ST 25), Guanyuan (CV 4), Zhongji (CV 3), Zigong (EX-CA 1), Liang qi u (ST 34), Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39) after menstruation until the ovulation stopped, and oral administration of progesterone was applied after ovulation. The patients in the control group three months before ET were treated with antibiotics in the menstrual period, and oral administration of progesterone was applied after ovulation. Continuous three menstrual periods were carried out for the both groups. The changes of endometrial thickness, type and endometrial blood flow and the outcome of FET were observed. Endometrial morphology and blood flow were improved after treatment in the two groups (all P Warm needling may improve endometrial receptivity, embryo transplantation rate and pregnancy rate and decrease early abortion rate by regulating endometrial morphology and blood flow.

  19. Generation of chimeric minipigs by aggregating 4- to 8-cell-stage blastomeres from somatic cell nuclear transfer with the tracing of enhanced green fluorescent protein.

    Science.gov (United States)

    Ji, Huili; Long, Chuan; Feng, Chong; Shi, Ningning; Jiang, Yingdi; Zeng, Guomin; Li, Xirui; Wu, Jingjing; Lu, Lin; Lu, Shengsheng; Pan, Dengke

    2017-05-01

    Blastocyst complementation is an important technique for generating chimeric organs in organ-deficient pigs, which holds great promise for solving the problem of a shortage of organs for human transplantation procedures. Porcine chimeras have been generated using embryonic germ cells, embryonic stem cells, and induced pluripotent stem cells; however, there are no authentic pluripotent stem cells for pigs. In previous studies, blastomeres from 4- to 8-cell-stage parthenogenetic embryos were able to generate chimeric fetuses efficiently, but the resulting fetuses did not produce live-born young. Here, we used early-stage embryos from somatic cell nuclear transfer (SCNT) to generate chimeric piglets by the aggregation method. Then, the distribution of chimerism in various tissues and organs was observed through the expression of enhanced green fluorescent protein (EGFP). Initially, we determined whether 4- to 8- or 8- to 16-cell-stage embryos were more suitable to generate chimeric piglets. Chimeras were produced by aggregating two EGFP-tagged Wuzhishan minipig (WZSP) SCNT embryos and two Bama minipig (BMP) SCNT embryos. The chimeric piglets were identified by coat color and microsatellite and swine leukocyte antigen analyses. Moreover, the distribution of chimerism in various tissues and organs of the piglets was evaluated by EGFP expression. We found that more aggregated embryos were produced using 4- to 8-cell-stage embryos (157/657, 23.9%) than 8- to 16-cell-stage embryos (100/499, 20.0%). Thus, 4- to 8-cell-stage embryos were used for the generation of chimeras. The rate of blastocysts development after aggregating WZSP with BMP embryos was 50.6%. Transfer of 391 blastocysts developed from 4- to 8-cell-stage embryos to five recipients gave rise to 18 piglets, of which two (11.1%) were confirmed to be chimeric by their coat color and microsatellite examination of the skin. One of the chimeric piglets died at 35 days and was subsequently autopsied, whereas the

  20. Intra- and interobserver analysis in the morphological assessment of early stage embryos during an IVF procedure: a multicentre study

    Directory of Open Access Journals (Sweden)

    Devroe Johanna

    2011-09-01

    Full Text Available Abstract Background Quality control programs are necessary to maintain good clinical practice. Embryo grading has been described as one of the external quality assurance schemes. Although the evaluation of embryos is based on the assessment of morphological characteristics, considerable intra- and inter-observer variability has been described. In this multicentre study, the variability in the embryo evaluation has been evaluated using morphological characteristics on day 1, day 2 and day 3 of embryo development. Methods Five embryologists of four different IVF centers participated in this study. Multilevel images of embryos were presented on a website at different time points to evaluate intra-and inter-observer agreement in the assessment of embryo morphology. The embryos were evaluated on day 1, day 2 and day 3 of their development and each embryologist had to decide if the embryo had to be transferred, cryopreserved or discarded. Results Both intra-observer agreement and inter-observer agreement were good to excellent for the position of the pronuclei on day 1, the number of blastomeres on day 2 and day 3 and the clinical decision (transfer, cryopreservation, discard. For all other characteristics (size of pronuclei, presence of cytoplasomic halo, degree of fragmentation and size of blastomeres the intra- and inter-observer agreement was moderate to very poor. Conclusions Mono- or multicentre quality control on embryo scoring by morphological assessment can easily be performed through the design of a simple website. In the future the website design can be adapted to generate statistical feedback upon scoring and can even include a training module.

  1. Dynamic changes in NuMA and microtubules in monkey-rabbit nuclear transfer embryos.

    Science.gov (United States)

    Yan, Li-Ying; Shi, Li-Hong; Sheng, Hui-Zhen; Liu, Shu-Zhen; Huang, Jun-Cheng; Zhu, Zi-Yu; OuYang, Ying-Chun; Lei, Zi-Li; Song, Xiang-Fen; Sun, Qing-Yuan; Chen, Da-Yuan

    2006-05-01

    Previous reports have indicated that failure in cloning monkey is attributed to the removal of nuclear mitotic apparatus (NuMA) during enucleation and subsequent abnormal organization of mitotic apparatus. This study investigated the transformation and assembly of tubulin and NuMA protein during the first cell cycle of cloned monkey embryos reconstructed by using enucleated rabbit oocytes as recipients. After the oocyte fused with a fibroblast, extensive microtubule organization was observed around the introduced nucleus in most reconstructed embryos, suggesting the introduction of a somatic cell centrosome. A high proportion of fibroblast nuclei transferred into non-activated oocytes underwent premature chromosome condensation (PCC), transient spindle organization and chromosomes separation, followed by the formation of two pronucleus-like structures. In contrast, fibroblast nuclei in pre-activated ooplasm rarely underwent PCC, but formed a swollen pronucleus-like structure. Normal spindles were observed in about one third of the cloned embryos reconstructed by both methods. After transferring monkey fibroblasts into NuMA-removed enucleated rabbit oocytes, NuMA was localized in pseudo-pronuclei and gradually moved to mitotic spindle poles at the first mitotic spindle poles. NuMA antibody microinjection resulted in spindle disorganization and chromosome misalignment, but did not significantly affect early cleavage. Our findings indicate that: 1. NuMA in donor monkey fibroblast may contribute to form a normal spindle in enucleated rabbit oocyte; 2. when non-activated cytoplasts and pre-activated cytoplasts are used as recipients, the donor nuclei undergo different morphological changes, but yield similar early embryo development; 3. although abnormal spindle organization and chromosome alignment may cause low efficiency of animal cloning, these abnormalities do not significantly affect early cleavage.

  2. Genome-Wide DNA Methylation Patterns of Bovine Blastocysts Developed In Vivo from Embryos Completed Different Stages of Development In Vitro.

    Directory of Open Access Journals (Sweden)

    Dessie Salilew-Wondim

    Full Text Available Early embryonic loss and altered gene expression in in vitro produced blastocysts are believed to be partly caused by aberrant DNA methylation. However, specific embryonic stage which is sensitive to in vitro culture conditions to alter the DNA methylation profile of the resulting blastocysts remained unclear. Therefore, the aim of this study was to investigate the stage specific effect of in vitro culture environment on the DNA methylation response of the resulting blastocysts. For this, embryos cultured in vitro until zygote (ZY, 4-cell (4C or 16-cell (16C were transferred to recipients and the blastocysts were recovery at day 7 of the estrous cycle. Another embryo group was cultured in vitro until blastocyst stage (IVP. Genome-wide DNA methylation profiles of ZY, 4C, 16C and IVP blastocyst groups were then determined with reference to blastocysts developed completely under in vivo condition (VO using EmbryoGENE DNA Methylation Array. To assess the contribution of methylation changes on gene expression patterns, the DNA methylation data was superimposed to the transcriptome profile data. The degree of DNA methylation dysregulation in the promoter and/or gene body regions of the resulting blastocysts was correlated with successive stages of development the embryos advanced under in vitro culture before transfer to the in vivo condition. Genomic enrichment analysis revealed that in 4C and 16C blastocyst groups, hypermethylated loci were outpacing the hypomethylated ones in intronic, exonic, promoter and proximal promoter regions, whereas the reverse was observed in ZY blastocyst group. However, in the IVP group, as much hypermethylated as hypomethylated probes were detected in gene body and promoter regions. In addition, gene ontology analysis indicated that differentially methylated regions were found to affected several biological functions including ATP binding in the ZY group, programmed cell death in the 4C, glycolysis in 16C and genetic

  3. EVALUATING THE EFFECTS OF FLY ASH EXPOSURE ON FISH EARLY LIFE STAGES: FATHEAD MINNOW EMBRYO-LARVAL TESTS

    Energy Technology Data Exchange (ETDEWEB)

    Greeley Jr, Mark Stephen [ORNL; Elmore, Logan R [ORNL; McCracken, Kitty [ORNL

    2012-05-01

    On December 22, 2008, a dike containing fly ash and bottom ash in an 84-acre complex of the Tennessee Valley Authority's (TVA) Kingston Steam Plant in East Tennessee failed and released a large quantity of ash into the adjacent Emory River. Ash deposits extended as far as 4 miles upstream (Emory River mile 6) of the Plant, and some ash was carried as far downstream as Tennessee River mile 564 ({approx}4 miles downstream of the Tennessee River confluence with the Clinch River). A byproduct of coal burning power plants, fly ash contains a variety of metals and other elements which, at sufficient concentrations and in specific forms, can be toxic to biological systems. The effects of fly ash contamination on exposed fish populations depend on the magnitude and duration of exposure, with the most significant risk considered to be the effects of specific ash constituents, especially selenium, on fish early life stages. Uptake by adult female fish of fly ash constituents through the food chain and subsequent maternal transfer of contaminants to the developing eggs is thought to be the primary route of selenium exposure to larval fish (Woock and others 1987, Coyle and others 1993, Lemly 1999, Moscatello and others 2006), but direct contact of the fertilized eggs and developing embryos to ash constituents in river water and sediments is also a potential risk factor (Woock and others 1987, Coyle and others 1993, Jezierska and others 2009). To address the risk of fly ash from the Kingston spill to the reproductive health of downstream fish populations, ORNL has undertaken a series of studies in collaboration with TVA including: (1) a field study of the bioaccumulation of fly ash constituents in fish ovaries and the reproductive condition of sentinel fish species in reaches of the Emory and Clinch Rivers affected by the fly ash spill; (2) laboratory tests of the potential toxicity of fly ash from the spill area on fish embryonic and larval development (reported in the

  4. Should we consider day-2 and day-3 embryo morphology before day-5 transfer when blastocysts reach a similar good quality?

    Science.gov (United States)

    Herbemont, Charlene; Sarandi, Solmaz; Boujenah, Jeremy; Cedrin-Durnerin, Isabelle; Sermondade, Nathalie; Vivot, Alexandre; Poncelet, Christophe; Grynberg, Michael; Sifer, Christophe

    2017-11-01

    Clinical outcomes of 291 day-5 blastocyst transfers carried out between January 2012 and March 2016 were retrospectively compared according to their quality at day 2 and 3. Inclusion criteria were female age younger than 37 years; first or second IVF and intracytoplasmic sperm injection cycle; quality of the transferred blastocyst: blastocoele B3 or higher; inner-cell-mass A/B; trophectoderm A/B; and known implantation outcome for each transferred blastocyst. Blastocysts were classified into good-quality and poor-quality embryo groups at day 2 and 3. Implantation (38.7% versus 41.4), clinical pregnancy (40.3% versus 45.9%), miscarriage (22.2% versus 26.7%;) and live birth rates (37.4% versus 38.8%) were comparable in day 2 good and poor-quality embryo groups. No signficiant differences in morphology of transferred blastocysts at day 3 were found. Multivariable analysis highlighted that poor or good embryo quality at day 2 and day 3 were not predictive of the implantation of good-quality blastocysts (at day 2: adjusted odds ratio = 0.82 CI 95% 0.49 to 1.38; at day 3: adjusted odds ratio = 1.39; CI 95% 0.77 to 2.52). Good-quality blastocyst transfer should, therefore, be carried out irrespective of embryo quality at cleavage stage, as it may not compromise success rates in a good-prognosis population. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  5. Initial stages of calcium uptake and mineral deposition in sea urchin embryos.

    Science.gov (United States)

    Vidavsky, Netta; Addadi, Sefi; Mahamid, Julia; Shimoni, Eyal; Ben-Ezra, David; Shpigel, Muki; Weiner, Steve; Addadi, Lia

    2014-01-07

    Sea urchin larvae have an endoskeleton consisting of two calcitic spicules. We reconstructed various stages of the formation pathway of calcium carbonate from calcium ions in sea water to mineral deposition and integration into the forming spicules. Monitoring calcium uptake with the fluorescent dye calcein shows that calcium ions first penetrate the embryo and later are deposited intracellularly. Surprisingly, calcium carbonate deposits are distributed widely all over the embryo, including in the primary mesenchyme cells and in the surface epithelial cells. Using cryo-SEM, we show that the intracellular calcium carbonate deposits are contained in vesicles of diameter 0.5-1.5 μm. Using the newly developed airSEM, which allows direct correlation between fluorescence and energy dispersive spectroscopy, we confirmed the presence of solid calcium carbonate in the vesicles. This mineral phase appears as aggregates of 20-30-nm nanospheres, consistent with amorphous calcium carbonate. The aggregates finally are introduced into the spicule compartment, where they integrate into the growing spicule.

  6. Initial stages of calcium uptake and mineral deposition in sea urchin embryos

    Science.gov (United States)

    Vidavsky, Netta; Addadi, Sefi; Mahamid, Julia; Shimoni, Eyal; Ben-Ezra, David; Shpigel, Muki; Weiner, Steve; Addadi, Lia

    2014-01-01

    Sea urchin larvae have an endoskeleton consisting of two calcitic spicules. We reconstructed various stages of the formation pathway of calcium carbonate from calcium ions in sea water to mineral deposition and integration into the forming spicules. Monitoring calcium uptake with the fluorescent dye calcein shows that calcium ions first penetrate the embryo and later are deposited intracellularly. Surprisingly, calcium carbonate deposits are distributed widely all over the embryo, including in the primary mesenchyme cells and in the surface epithelial cells. Using cryo-SEM, we show that the intracellular calcium carbonate deposits are contained in vesicles of diameter 0.5–1.5 μm. Using the newly developed airSEM, which allows direct correlation between fluorescence and energy dispersive spectroscopy, we confirmed the presence of solid calcium carbonate in the vesicles. This mineral phase appears as aggregates of 20–30-nm nanospheres, consistent with amorphous calcium carbonate. The aggregates finally are introduced into the spicule compartment, where they integrate into the growing spicule. PMID:24344263

  7. Development of interspecies cloned embryos in yak and dog.

    Science.gov (United States)

    Murakami, Masao; Otoi, Takeshige; Wongsrikeao, Pimprapar; Agung, Budiyanto; Sambuu, Rentsenkhand; Suzuki, Tatsuyuki

    2005-01-01

    Interspecies nuclear transfer (NT) could be an alternative to replicate animals when supply of recipient oocytes is limited or in vitro embryo production systems are incomplete. In the present study, embryonic development was assessed following interspecies NT of donor cumulus cells derived from yak and dog into the recipient ooplasm of domestic cow. The percentages of fusion and subsequent embryo development to the eight-cell stage of interspecies NT embryos were comparable to those of intraspecies NT embryos (cow-cow NT embryos). The percentage of development to blastocysts was significantly lower (p dog-cow NT embryos, only one embryo (0.4%) developed to the blastocyst stage. These results indicate that interspecies NT embryos possess equally developmental competence to the eight-cell stage as intraspecies NT embryos, but the development to blastocysts is very low when dog somatic cells are used as the donor nuclei.

  8. Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos

    International Nuclear Information System (INIS)

    Zhong Zhisheng; Zhang Gang; Meng Xiaoqian; Zhang Yanling; Chen Dayuan; Schatten, Heide; Sun Qingyuan

    2005-01-01

    Centrosomes, the main microtubule-organizing centers (MTOCs) in most animal cells, are important for many cellular activities such as assembly of the mitotic spindle, establishment of cell polarity, and cell movement. In nuclear transfer (NT), MTOCs that are located at the poles of the meiotic spindle are removed from the recipient oocyte, while the centrosome of the donor cell is introduced. We used mouse MII oocytes as recipients, mouse fibroblasts, rat fibroblasts, or pig granulosa cells as donor cells to construct intraspecies and interspecies nuclear transfer embryos in order to observe centrosome dynamics and functions. Three antibodies against centrin, γ-tubulin, and NuMA, respectively, were used to stain the centrosome. Centrin was not detected either at the poles of transient spindles or at the poles of first mitotic spindles. γ-tubulin translocated into the two poles of the transient spindles, while no accumulated γ-tubulin aggregates were detected in the area adjacent to the two pseudo-pronuclei. At first mitotic metaphase, γ-tubulin was translocated to the spindle poles. The distribution of γ-tubulin was similar in mouse intraspecies and rat-mouse interspecies embryos. The NuMA antibody that we used can recognize porcine but not murine NuMA protein, so it was used to trace the NuMA protein of donor cell in reconstructed embryos. In the pig-mouse interspecies reconstructed embryos, NuMA concentrated between the disarrayed chromosomes soon after activation and translocated to the transient spindle poles. NuMA then immigrated into pseudo-pronuclei. After pseudo-pronuclear envelope breakdown, NuMA was located between the chromosomes and then translocated to the spindle poles of first mitotic metaphase. γ-tubulin antibody microinjection resulted in spindle disorganization and retardation of the first cell division. NuMA antibody microinjection also resulted in spindle disorganization. Our findings indicate that (1) the donor cell centrosome, defined as

  9. Experimental risk assessment of bovine viral diarrhea virus transmission via in vitro embryo production using somatic cell nucleus transfer.

    Science.gov (United States)

    Gregg, K; Chen, S H; Sadeghieh, S; Guerra, T; Xiang, T; Meredith, J; Polejaeva, I

    2009-07-01

    The objective of this study was to perform a comprehensive risk assessment on infectious disease transmission in the system of in vitro embryo production via somatic cell nucleus transfer (SCNT) technology using bovine viral diarrhea virus (BVDV) as a model. The risks of BVDV transmission in each step of the SCNT embryo production procedure, from donor cells to preimplantation SCNT embryo culture, were carefully examined using a sensitive real-time polymerase chain reaction assay. The identified primary source of BVDV transmission in SCNT embryo production was donor cell infection, most likely caused by contaminated fetal bovine serum in the culture medium. The risk of disease transmission through contaminated oocytes from an abattoir was relatively low, and it can be greatly minimized by cumulus cell removal and adequate oocyte washing procedures. Of the 200 cumulus-oocyte complexes (COCs) and more than 1500 cumulus cell-free oocyte (CFO) samples collected from multiple sources over a course of 7 months, only 2.5% of the COCs were BVDV positive, and all of the CFOs (100%) were BVDV negative. To evaluate the risk of BVDV introduction during in vitro SCNT embryo culture, 324 SCNT embryos were produced from 18 different cell lines using oocytes from 26 different batches collected over a course of 9 months. The embryos were cultured in vitro for 7 days and then tested for BVDV. All of the 324 SCNT embryos (100%) were negative, indicating that the embryo culture system is virtually risk-free for BVDV transmission. Based on these results, a standard operational protocol (SOP) for SCNT embryo production was proposed to greatly minimize the risk of BVDV transmission through the SCNT embryo production system. This SOP could be a starting point to produce a SCNT system that is virtually risk-free for disease transmission in general.

  10. Assisted reproductive technology use, embryo transfer practices, and birth outcomes after infertility insurance mandates: New Jersey and Connecticut.

    Science.gov (United States)

    Crawford, Sara; Boulet, Sheree L; Jamieson, Denise J; Stone, Carol; Mullen, Jewel; Kissin, Dmitry M

    2016-02-01

    To explore whether recently enacted infertility mandates including coverage for assisted reproductive technology (ART) treatment in New Jersey (2001) and Connecticut (2005) increased ART use, improved embryo transfer practices, and decreased multiple birth rates. Retrospective cohort study using data from the National ART Surveillance System. We explored trends in ART use, embryo transfer practices and birth outcomes, and compared changes in practices and outcomes during a 2-year period before and after passing the mandate between mandate and non-mandate states. Not applicable. Cycles of ART performed in the United States between 1996 and 2013. Infertility insurance mandates including coverage for ART treatment passed in New Jersey (2001) and Connecticut (2005). Number of ART cycles performed, number of embryos transferred, multiple live birth rates. Both New Jersey and Connecticut experienced an increase in ART use greater than the non-mandate states. The mean number of embryos transferred decreased significantly in New Jersey and Connecticut; however, the magnitudes were not significantly different from non-mandate states. There was no significant change in ART birth outcomes in either mandate state except for an increase in live births in Connecticut; the magnitude was not different from non-mandate states. The infertility insurance mandates passed in New Jersey and Connecticut were associated with increased ART treatment use but not a decrease in the number of embryos transferred or the rate of multiples; however, applicability of the mandates was limited. Published by Elsevier Inc.

  11. Endometrial thickness significantly affects clinical pregnancy and live birth rates in frozen-thawed embryo transfer cycles.

    Science.gov (United States)

    Bu, Zhiqin; Wang, Keyan; Dai, Wei; Sun, Yingpu

    2016-07-01

    In order to explore the relationship between endometrial thickness on the day of embryo transfer and pregnancy outcomes in frozen-thawed embryo transfer (FET) cycles, we retrospectively analyzed data from 2997 patients undergoing their first FET cycles from January 2010 to December 2012. All patients were divided into three groups (Group A, ≤8 mm; Group B, 9-13 mm; Group C, ≥14 mm) according to the endometrial thickness on embryo transfer day. Compared with patients in the other two groups, patients with thin endometrial thickness in Group A had significantly lower clinical pregnancy rate (33.4%, 41.3% and 45.4%, p birth rate (23.8%, 32.2% and 34.0%, p confidence interval (CI): 1.10-1.77, p birth rate (aOR: 1.50; 95% CI: 1.16-1.95, p < 0.01) were significant. We conclude that for patients undergoing FET, endometrial thickness on the embryo transfer day significantly affects IVF outcomes in cleavage embryo transfer cycles independent of other factors.

  12. Effects of donor fibroblast cell type and transferred cloned embryo number on the efficiency of pig cloning.

    Science.gov (United States)

    Li, Zicong; Shi, Junsong; Liu, Dewu; Zhou, Rong; Zeng, Haiyu; Zhou, Xiu; Mai, Ranbiao; Zeng, Shaofen; Luo, Lvhua; Yu, Wanxian; Zhang, Shouquan; Wu, Zhenfang

    2013-02-01

    Currently, cloning efficiency in pigs is very low. Donor cell type and number of cloned embryos transferred to an individual surrogate are two major factors that affect the successful rate of somatic cell nuclear transfer (SCNT) in pigs. This study aimed to compare the influence of different donor fibroblast cell types and different transferred embryo numbers on recipients' pregnancy rate and delivery rate, the average number of total clones born, clones born alive and clones born healthy per litter, and the birth rate of healthy clones (=total number of healthy cloned piglets born /total number of transferred cloned embryos). Three types of donor fibroblasts were tested in large-scale production of cloned pigs, including fetal fibroblasts (FFBs) from four genetically similar Western swine breeds of Pietrain (P), Duroc (D), Landrace (L), and Yorkshire (Y), which are referred to as P,D,LY-FFBs, adult fibroblasts (AFBs) from the same four breeds, which are designated P,D,L,Y-AFBs, and AFBs from a Chinese pig breed of Laiwu (LW), which is referred to as LW-AFBs. Within each donor fibroblast cell type group, five transferred cloned embryo number groups were tested. In each embryo number group, 150-199, 200-249, 250-299, 300-349, or 350-450 cloned embryos were transferred to each individual recipient sow. For the entire experiment, 92,005 cloned embryos were generated from nearly 115,000 matured oocytes and transferred to 328 recipients; in total, 488 cloned piglets were produced. The results showed that the mean clones born healthy per litter resulted from transfer of embryos cloned from LW-AFBs (2.53 ± 0.34) was similar with that associated with P,D,L,Y-FFBs (2.72 ± 0.29), but was significantly higher than that resulted from P,D,L,Y-AFBs (1.47 ± 0.18). Use of LW-AFBs as donor cells for SCNT resulted in a significantly higher pregnancy rate (72.00% vs. 59.30% and 48.11%) and delivery rate (60.00% vs. 45.93% and 35.85%) for cloned embryo recipients, and a

  13. Manipulating early pig embryos.

    Science.gov (United States)

    Niemann, H; Reichelt, B

    1993-01-01

    On the basis of established surgical procedures for embryo recovery and transfer, the early pig embryo can be subjected to various manipulations aimed at a long-term preservation of genetic material, the generation of identical multiplets, the early determination of sex or the alteration of the genetic make-up. Most of these procedures are still at an experimental stage and despite recent considerable progress are far from practical application. Normal piglets have been obtained after cryopreservation of pig blastocysts hatched in vitro, whereas all attempts to freeze embryos with intact zona pellucida have been unsuccessful. Pig embryos at the morula and blastocyst stage can be bisected microsurgically and the resulting demi-embryos possess a high developmental potential in vitro, whereas their development in vivo is impaired. Pregnancy rates are similar (80%) but litter size is reduced compared with intact embryos and twinning rate is approximately 2%. Pig blastomeres isolated from embryos up to the 16-cell stage can be grown in culture and result in normal blastocysts. Normal piglets have been born upon transfer of blastocysts derived from isolated eight-cell blastomeres, clearly underlining the totipotency of this developmental stage. Upon nuclear transfer the developmental capacity of reconstituted pig embryos is low and culture. Sex determination can be achieved either by separation of X and Y chromosome bearing spermatozoa by flow cytometry or by analysing the expression of the HY antigen in pig embryos from the eight-cell to morula stage. Microinjection of foreign DNA has been successfully used to alter growth and development of transgenic pigs, and to produce foreign proteins in the mammary gland or in the bloodstream, indicating that pigs can be used as donors for valuable human pharmaceutical proteins. Another promising area of gene transfer is the increase of disease resistance in transgenic lines of pigs. Approximately 30% of pig spermatozoa bind

  14. Evaluation of the effect of indomethacin and piroxicam administration before embryo transfer on pregnancy rate.

    Science.gov (United States)

    Kumbasar, Serkan; Gül, Özer; Şık, Aytek

    2017-03-01

    The aim of this study was to evaluate the effect of non-steroidal anti-inflammatory drug (NSAID) administration before embryo transfer (ET) on pregnancy rates in women undergoing in vitro fertilization/intracytoplasmic sperm injection ET. Our study included 255 patients diagnosed with primary or secondary infertility caused by a male or tubal-related factor, endometriosis or unexplained factors. The patients were divided randomly into three groups. Two groups were administered oral piroxicam (10 mg capsules) or 100 mg indomethacin (rectal suppository), respectively, 1-2 h before ET. As a control, the third group did not receive any form of treatment before ET. Basal levels of follicle-stimulating hormone, luteinizing hormone, and level 17β-estradiol on the day of human chorionic gonadotropin administration, the collected and transferred number of embryos, and the number of grade A embryos obtained were determined in all patients. The implantation, clinical pregnancy, and miscarriage rates of the groups were compared. The clinical pregnancy rate per ET and the implantation rate were 35.2% and 12.15% in the piroxicam group, 31.7% and 10.9% in the indomethacin group, and 32.9% and 12.5% in the control, respectively. The miscarriage rates of groups 1, 2 and 3 were 12%, 11.7% and 11.7%, respectively (P = 0.964). The differences in clinical pregnancy rates among the groups were not statistically significant (P = 0.887). There were also no significant differences in the implantation rates (P = 0.842). These results suggest that NSAID administration before ET has no additional effect on pregnancy outcome in patients undergoing in vitro fertilization. © 2017 Japan Society of Obstetrics and Gynecology.

  15. Factors that Influence The Occurrence of Multiple Pregnancies after Intracytoplasmic Injection Cycles with Two or Three Fresh Embryo Transfers.

    Science.gov (United States)

    Abdollahi, Mahbubeh; Omani Samani, Reza; Hemat, Mandana; Arabipoor, Arezoo; Shabani, Fatemeh; Eskandari, Farzad; Salehi, Masoud

    2017-10-01

    Multiple pregnancies are an important complication of assisted reproductive technology (ART). The present study aims to indentify the risk factors for multiple pregnancies independent of the number of transferred embryos. This retrospective study reviewed the medical records of patients who underwent intracytoplasmic sperm injection (ICSI) cycles in Royan Institute between October 2011 and January 2012. We entered 12 factors that affected the number of gestational sacs into the poisson regression (PR) model. Factors were obtained from two study populations-cycles with double embryo transfer (DET) and cycles that transferred three embryos (TET). We sought to determine the factors that influenced the number of gestational sacs. These factors were entered into multivariable logistic regression (MLR) to identify risk factors for multiple pregnancies. A total of 1000 patients referred to Royan Institute for ART during the study period. We included 606 eligible patients in this study. PR analysis demonstrated that the quality of transferred embryos and woman's age had a significant effect on the number of observed sacs in patients who underwent ICSI with DET. There was no significant predictive variable for multiple pregnancies according to MLR analysis. Our findings demonstrated that both regression models (PR and MLR) had the same outputs. A significant relation existed between age and fertilization rate with multiple pregnancies in patients who underwent ICSI with TET. Single embryo transfer (SET) should be considered with the remaining embryos cryopreserved to prevent multiple pregnancies in women younger than 35 years of age who undergo ICSI cycles with high fertilization rates and good or excellent quality embryos. However, further prospective studies are necessary to evaluate whether SET in women with these risk factors can significantly decrease multiple pregnancies and improve cycle outcomes. Copyright© by Royan Institute. All rights reserved.

  16. Pregnancy, Delivery, and Neonatal Outcomes of In Vitro Fertilization-Embryo Transfer in Patient with Previous Cesarean Scar

    Science.gov (United States)

    Zhang, Ningyuan; Chen, Hua; Xu, Zhipeng; Wang, Bin; Sun, Haixiang; Hu, Yali

    2016-01-01

    Background What role should previous cesarean section play in affecting clinical pregnancy outcomes and avoiding the complications of in vitro fertilization? In this article, we focus on elective single-embryo transfer (eSET) versus double-embryo transfer (DET) and assess the clinical efficacy and safety of eSET in patients who have a previous cesarean scar. Material/Methods The pregnancy, delivery, and neonatal outcomes of 130 patients who had a previous cesarean scar and received in vitro fertilization-embryo transfer (IVF-ET) were retrospectively analyzed. The number of transferred embryos was chosen depending on patients’ desire after acknowledging all benefits and risks, including eSET (eSET group, n=56) and DET (DET group, n=74). A total of 101 patients with previous vaginal delivery receiving IVF-ET in the same period were included as a control group. Results The pregnancy rates, multiple birth rates, abortion rates, ectopic pregnancy rates, gestational age at delivery, preterm birth rates, neonatal birth weight, and take-home baby rates were similar between the previous cesarean section group and the previous vaginal delivery group. A previous cesarean section scar did not affect embryo implantation and pregnancy outcomes in IVF. In the eSET and DET groups of previous cesarean section patients, the embryo implantation rates, pregnancy rates, abortion rates, and take-home baby rates were similar. However, the rate of multiple pregnancies reached 50% in the DET group, which led to more preterm births and lower birth weight. Conclusions Elective single-embryo transfer is a well-accepted strategy to avoid multiple pregnancies and improve the obstetric and neonatal outcomes of singleton pregnancy in IVF patients with a previous cesarean section. PMID:27636504

  17. Pregnancy loss after frozen-embryo transfer--a comparison of three protocols

    DEFF Research Database (Denmark)

    Tomás, Candido; Alsbjerg, Birgit; Martikainen, Hannu

    2012-01-01

    of embryos and ET. MAIN OUTCOME MEASURE(S): Pregnancy test rate, clinical pregnancy rate, and pregnancy loss rate. RESULT(S): The natural cycle followed by P (NC + P) was used in 26% of cycles, the natural cycle with hCG (NC + hCG) in 10%, and the substituted cycle with estrogen and P (E + P) in 64......, irregular cycles, endometrial thickness, number, and quality of embryos transferred did not correlate to pregnancy loss. CONCLUSION(S): A higher positive pregnancy test rate was obtained in E + P frozen ET cycles in comparison with other protocols; however, due to an increased preclinical and clinical......% of cycles. The rate of transfers after thawing was similar in all groups (87.2%, 73.9%, and 87.2%, respectively). There was a significantly higher positive pregnancy test rate in the E + P (34.3%) and NC + hCG (35.5%) cycles as compared with the NC + P cycles (26.7%). However, the clinical pregnancy rate...

  18. Generating Transgenic Mice by Lentiviral Transduction of Spermatozoa Followed by In Vitro Fertilization and Embryo Transfer.

    Science.gov (United States)

    Chandrashekran, Anil; Casimir, Colin; Dibb, Nick; Readhead, Carol; Winston, Robert

    2016-01-01

    Most transgenic technologies rely on the oocyte as a substrate for genetic modification. Transgenics animals are usually generated by the injection of the gene constructs (including lentiviruses encoding gene constructs or modified embryonic stem cells) into the pronucleus of a fertilized egg followed by the transfer of the injected embryos into the uterus of a foster mother. Male germ cells also have potential as templates for transgenic development. We have previously shown that mature sperm can be utilized as template for lentiviral transduction and as such used to generate transgenic mice efficiently with germ line capabilities. We provide here a detailed protocol that is relatively simple, to establish transgenic mice using lentivirally transduced spermatozoa. This protocol employs a well-established lentiviral gene delivery system (usual for somatic cells) delivering a variety of transgenes to be directly used with sperm, and the subsequent use of these modified sperm in in vitro fertilization studies and embryo transfer into foster female mice, for the establishment of transgenic mice.

  19. Perinatal outcome in fresh versus frozen embryo transfer in ART cycles

    Directory of Open Access Journals (Sweden)

    Ali Aflatoonian

    2016-03-01

    Full Text Available Background: Despite of higher pregnancy rate after frozen embryo transfer (FET which is accepted by the majority of researches, the safety of this method and its effect on neonatal outcome is still under debate. Objective: The aim of this study was to evaluate pregnancy and neonatal outcome of FET compare to fresh cycles. Materials and Methods: In this study,1134 patients using fresh ET and 285 women underwent FET were investigated regarding live birth as primary outcome and gestational age, birth weight, gender, multiple status, ectopic pregnancy, still birth and pregnancy loss as secondary outcomes. Results: Our results showed that there is no difference between FET and fresh cycles regarding live birth (65.6% vs. 70.4% respectively. Ectopic pregnancy, still birth and abortion were similar in both groups. The mean gestational age was significantly lower among singletons in FET group compared to fresh cycles (p=0.047. Prematurity was significantly elevated among singleton infants in FET group (19.6% in comparison to neonates born after fresh ET (12.8% (p=0.037. Conclusion: It seems that there is no major difference regarding perinatal outcome between fresh and frozen embryo transfer. Although, live birth is slightly increased in fresh cycles and prematurity was significantly increased among singleton infants in FET group

  20. Perinatal outcome after embryo transfer in ovum recipients. A comparison with standard in vitro fertilization.

    Science.gov (United States)

    Friedman, F; Copperman, A B; Brodman, M L; Shah, D; Sandler, B; Grunfeld, L

    1996-09-01

    To compare perinatal outcome after embryo transfer vs. standard in vitro fertilization (IVF) in ovum recipients. We reviewed 22 consecutive ovum donor pregnancies delivered at Mount Sinai Hospital between July 1989 and November 1992 and matched them for age, parity and order of gestation to a control group who underwent standard IVF-embryo transfer (ET) during that period. The two groups showed similar rates of maternal complications, including hypertensive disorders, gestational diabetes and puerperal complications. When compared to patients undergoing autologous IVF-ET, ovum recipients had a lower rate of preterm labor (22.7% vs. 54.6%, P < .05), a higher mean gestational age at delivery (38.7 vs. 36.1 weeks, P < .01) and increased mean birth weight of neonates (2,924 vs. 2,374 g, P < .005). Though infants born through traditional IVF-ET were more likely to be preterm, Apgar scores were similar at one and five minutes. Our data support the generally favorable outcome seen in pregnancies conceived through ovum donation despite the fact that many of these patients had prior poor prenatal outcomes, long intervals until conception, and coexisting medical conditions. Ovum donation in women of comparable ages has obstetric outcomes equal to or better than patients undergoing traditional IVF-ET. Obstetric and perinatal outcome do not seem to be impaired in patients receiving donated oocytes.

  1. Endometrial preparation: effect of estrogen dose and administration route on reproductive outcomes in oocyte donation cycles with fresh embryo transfer.

    Science.gov (United States)

    Madero, S; Rodriguez, A; Vassena, R; Vernaeve, V

    2016-08-01

    Is there a difference in live birth rates following endometrial preparation with either a constant or increasing estrogen dose in fresh embryo transfer from oocyte donation cycles? There is no difference in live birth rates between a constant dose versus an increasing dose of estrogen after fresh embryo transfer in oocyte donation cycles with oral or transdermal supplementation. Endometrial preparation (EP) with estrogen and progesterone, and embryo-endometrial synchronicity are determinant for adequate embryo implantation. Estrogen is crucial and different exogenous administration patterns could imply variations on EP. Moreover, estrogen undergoes metabolization by the intestines and liver when administered orally, an effect that is bypassed by transdermal administration. Information on the effect of replacement patterns and route of administration of E on reproductive outcomes of women undergoing fresh embryo transfer from oocyte donation cycles is scarce. Retrospective cohort study including 8362 embryo transfers following ICSI, corresponding to 8254 patients, between October 2010 and March 2015. A total of 5593 (66.9%) patients received an increasing E dose (ID) (oral: 2 mg/day day(d)1-7, 4 mg days d8-12, 6 mg d13-embryo transfer; transdermal: 75 µg/3 days on d1-6, 150 µg/3 days d7-embryo transfer) while 2769 (33.1%) received a constant dose (CD) of estrogen (oral: 6 mg/day 1-embryo transfer; transdermal: 150 µg/3 days d1-embryo transfer). Embryos were generated by ICSI with fresh or vitrified donor oocytes fertilized with either fresh or frozen sperm from either the couple partner or donor. Cohort allocation was not related to patient characteristics; instead it reflected an internal policy change in E administration. Effect of estrogen dose (ID versus CD) on biochemical, clinical, ongoing and live birth rates, stratified by administration route, was analyzed by univariate and multivariate analysis adjusted by donor and recipient demographic and cycle

  2. Developmental toxicity of deltamethrin and 3-phenoxybenzoic acid in embryo-larval stages of zebrafish (Danio rerio).

    Science.gov (United States)

    Kuder, Reshma Shabnam; Gundala, Harold Philip

    2018-03-05

    Synthetic pyrethroids are the major insecticides used widely in agriculture and household pest control. Deltamethrin (DM), a widely used type II pyrethroid insecticide, is a relatively potent neurotoxicant. 3-Phenoxybenzoic acid (PBA) is the major metabolite formed due to metabolism of DM. In order to illustrate the toxic response of zebrafish embryos/larvae to DM and PBA the present research was carried out. For this 4hpf embryos were treated with two concentrations of DM (100 and 200 μg/L) for 48 h and PBA (1000 and 2000 μg/L) for 96 h or 99.9% ethanol (solvent control). Early life stage parameters were observed at specified time points. DM-treated embryo/larvae exhibited increased mortality, delay in hatching time, decrease in percentage of hatched embryos, increase of heartbeat rate and decrease in blood flow; lightening of body and eye pigmentation in a dose dependent manner. Pericardial and yolk sac edema along with were also caused by DM. Along with these crooked notochord, tail deformation was noticed in hatched and unhatched embryos. In case of PBA treated embryos and larvae, increased embryos/larvae length and yolk sac size were observed. Other abnormalities like edema (yolk sac and pericardial), decreased eye and body pigmentation were also observed but in some embryos only. These were not as severe as observed in parental compound indicating that DM is more toxic than its metabolite PBA. The data contributes to a better understanding of the potential consequences of fish exposed to DM and PBA.

  3. Current Status of Comprehensive Chromosome Screening for Elective Single-Embryo Transfer

    Directory of Open Access Journals (Sweden)

    Ming-Yih Wu

    2014-01-01

    Full Text Available Most in vitro fertilization (IVF experts and infertility patients agree that the most ideal assisted reproductive technology (ART outcome is to have a healthy, full-term singleton born. To this end, the most reliable policy is the single-embryo transfer (SET. However, unsatisfactory results in IVF may result from plenty of factors, in which aneuploidy associated with advanced maternal age is a major hurdle. Throughout the past few years, we have got a big leap in advancement of the genetic screening of embryos on aneuploidy, translocation, or mutations. This facilitates a higher success rate in IVF accompanied by the policy of elective SET (eSET. As the cost is lowering while the scale of genome characterization continues to be up over the recent years, the contemporary technologies on trophectoderm biopsy and freezing-thaw, comprehensive chromosome screening (CCS with eSET appear to be getting more and more popular for modern IVF centers. Furthermore, evidence has showen that, by these avant-garde techniques (trophectoderm biopsy, vitrification, and CCS, older infertile women with the help of eSET may have an opportunity to increase the success of their live birth rates approaching those reported in younger infertility patients.

  4. A Comparison of Success Rates of Embryo Transfer on Weekdays and Weekends

    Directory of Open Access Journals (Sweden)

    Pinar Solmaz Hasdemir

    2016-05-01

    Full Text Available Background: The aim of this study is to examine the effect of the embryo transfer (ET day on clinical pregnancy success rates in in vitro fertilization-ET (IVF-ET cycles. Materials and Methods: In this retrospective study, we divided patients with infertility who underwent IVF-ET with fresh embryos into two groups depending on whether the ET was performed on weekdays or weekends. The main outcome measure was to compare the clinical pregnancy rates of patients with similar demographic and clinical characteristics who underwent ET on weekdays or weekends. Results: A total of 188 patients underwent IVF-ET on weekdays (n=156 or weekends (n=32. Both groups had similar demographic and cycle characteristics. The overall pregnancy rate was 42.8%. Among the study groups, the weekday group had a 40.2% ET success rate and the weekend group had a 54.8% success rate (P=0.517. Although no statistically significant difference existed between the two groups, we observed an absolute 14.6% increase in pregnancy rate for ETs performed during weekends compared to those performed on weekdays, with a 35% statistical power. Conclusion: ETs performed during weekends were more successful than ETs performed during weekdays with an absolute 14.6% increase in clinical pregnancy rate. This finding should be confirmed by conducting further studies with larger groups of patients.

  5. Morphological characterization of pre- and peri-implantation in vitro cultured, somatic cell nuclear transfer and in vivo derived ovine embryos

    DEFF Research Database (Denmark)

    Tveden-Nyborg, Pernille Yde; Peura, T.T.; Hartwich, K.M.

    2005-01-01

    were produced by in vitro culture of in vivo fertilized zygotes. Most SCNT and IVC embryos were transferred to recipients on day 6 while some remained in culture for day 7 processing. In vivo embryos were collected as zygotes, transferred to intermediate recipients and retransferred to final recipients...

  6. Nuclear and nuclear reprogramming during the first cell cycle in bovine nuclear transfer embryos

    DEFF Research Database (Denmark)

    Østrup, Olga; Petrovicova, Ida; Strejcek, Frantisek

    2009-01-01

    Abstract The immediate events of genomic reprogramming at somatic cell nuclear transfer (SCNT) are to high degree unknown. This study was designed to evaluate the nuclear and nucleolar changes during the first cell cycle. Bovine SCNT embryos were produced from starved bovine fibroblasts and fixed......, somatic cell nuclei introduced into enucleated oocytes displayed chromatin condensation, partial nuclear envelope breakdown, nucleolar desegregation and transcriptional quiescence already at 0.5 hpa. Somatic cell cytoplasm remained temporally attached to introduced nucleus and nucleolus was partially...... restored indicating somatic influence in the early SCNT phases. At 1-3 hpa, chromatin gradually decondensed toward the nucleus periphery and nuclear envelope reformed. From 4 hpa, the somatic cell nucleus gained a PN-like appearance and displayed NPBs suggesting ooplasmic control of development....

  7. Abnormal human chorionic gonadotropin (hCG) trends after transfer of multiple embryos resulting in viable singleton pregnancies.

    Science.gov (United States)

    Brady, Paula C; Farland, Leslie V; Missmer, Stacey A; Racowsky, Catherine; Fox, Janis H

    2017-12-19

    The purpose of this study is to investigate whether abnormal hCG trends occur at a higher incidence among women conceiving singleton pregnancies following transfer of multiple (two or more) embryos (MET), as compared to those having a single embryo transfer (SET). Retrospective cohort study was performed of women who conceived singleton pregnancies following fresh or frozen autologous IVF/ICSI cycles with day 3 or day 5 embryo transfers between 2007 and 2014 at a single academic medical center. Cycles resulting in one gestational sac on ultrasound followed by singleton live birth beyond 24 weeks of gestation were included. Logistic regression models adjusted a priori for patient age at oocyte retrieval and day of embryo transfer were used to estimate the Odds Ratio of having an abnormal hCG rise (defined as a rise or hCG rises between the first and second measurements, compared to 2.7% (n = 17) of patients undergoing SET (OR 2.16, 95% CI 1.26-3.71). Among patients with initially abnormal hCG rises who had a third level checked (89%), three-quarters had normal hCG rises between the second and third measurements. Patients who deliver singletons following MET were more likely to have suboptimal initial hCG rises, potentially due to transient implantation of other non-viable embryo(s). While useful for counseling, these findings should not change standard management of abnormal hCG rises following IVF. The third hCG measurements may clarify pregnancy prognosis.

  8. Thermal effect on heart rate and hemodynamics in vitelline arteries of stage 18 chicken embryos.

    Science.gov (United States)

    Lee, Jung Yeop; Lee, Sang Joon

    2010-12-01

    We investigated the thermal effects on heart rate, hemodynamics, and response of vitelline arteries of stage-18 chicken embryos. Heart rate was monitored by a high-speed imaging method, while hemodynamic quantities were evaluated using a particle image velocimetry (PIV) technique. Experiments were carried out at seven different temperatures (36-42 °C with 1 °C interval) after 1h of incubation to stabilize the heart rate. The heart rate increased in a linear manner (r = 0.992). Due to the increased cardiac output (or heart rate), the hemodynamic quantities such as mean velocity (U(mean)), velocity fluctuation (U(fluc)), and peak velocity (U(peak)) also increased with respect to the Womersley number (Ω) in the manner r = 0.599, 0.693, and 0.725, respectively. This indicates that the mechanical force exerting on the vessel walls increases. However, the active response (or regulation) of the vitelline arteries was not observed in this study. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Somatic cell count and type of intramammary infection impacts fertility from in vitro produced embryo transfer.

    Science.gov (United States)

    Barbosa, L F S P; Oliveira, W V C; Pereira, M H C; Moreira, M B; Vasconcelos, C G C; Silper, B F; Cerri, R L A; Vasconcelos, J L M

    2018-03-01

    The objective of this study was to assess the impact of mastitis-causing bacteria and somatic cell count (SCC) on pregnancy per embryo transfer (P/ET) in Holstein-Gir crossbred (Girolando) lactating dairy cows. Cows (n = 1397) were subjected to a timed-embryo transfer protocol. Milk samples were collected two days before embryo transfer for SCC and bacteriological culture analyses. Pregnancy diagnosis was performed on days 31 and 66 after timed-embryo transfer. The animals were grouped according to the National Mastitis Council recommendations: Gram-positive environmental (EV+), Gram-negative environmental (EV-), Gram-positive contagious (C+), coagulase-negative staphylococci (CNS) and control (no bacterial growth). Additional analysis was made by categorizing bacteria based on degree of pathogenicity (Major or Minor). Bacterial growth reduced P/ET (P  400,000 cells/mL had lower P/ET (P < .01) than animals with SCC < 200,000 cells/mL at both 31 (30.4% vs 40.8%) and 66 days (24.7% vs 32.2%) of gestation. Pregnancy loss was not different between bacterial isolates and SCC categories. Elevated SCC significantly reduced P/ET, whereas environmental agents and those with Major pathogenicity yielded the greatest reduction in P/ET. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Evaluation of genetic components in traits related to superovulation, in vitro fertilization, and embryo transfer in Holstein cattle

    Science.gov (United States)

    The objectives of this study were to estimate variance components and identify regions of the genome associated with traits related to embryo transfer in Holsteins. Reproductive technologies are used in the dairy industry to increase the reproductive rate of superior females. A drawback of these met...

  11. Evaluating the interaction between progesterone, TNF alpha and cortisol on early loss of transferred embryos in beef cows

    Science.gov (United States)

    Fifty-eight non-lactating cows previously synchronized for estrus were assigned to two treatments to assess the effects of progesterone supplementation and its correlation with TNF-a and cortisol on the survival of the transferred embryos. On day 7 after exhibiting estrus (day 0), cows in both group...

  12. Evaluation of an effective multifaceted implementation strategy for elective single-embryo transfer after in vitro fertilization

    NARCIS (Netherlands)

    Kreuwel, I.A.M.; Peperstraten, A.M. van; Hulscher, M.E.J.L.; Kremer, J.A.M.; Grol, R.P.T.M.; Nelen, W.L.D.M.; Hermens, R.P.M.G.

    2013-01-01

    STUDY QUESTION: What is the relationship between the rate of elective single-embryo transfer (eSET) and couples' exposure to different elements of a multifaceted implementation strategy? SUMMARY ANSWER: Additional elements in a multifaceted implementation strategy do not result in an increased eSET

  13. Characteristics of pregnancies and offspring following transfer of bovine in vivo embryos assessed by nanorespirometry

    DEFF Research Database (Denmark)

    Lopes, Ana Sofia; Madsen, S E; Greve, Torben

    2008-01-01

    .05), with quality I embryos associated with heavier calves when compared to quality III embryos. Furthermore, light traction was more frequently used to retrieve heavier animals (P male calves were 3.2 kg heavier than females (P

  14. Abdominal ectopic pregnancy after in vitro fertilization and single embryo transfer: a case report and systematic review.

    Science.gov (United States)

    Yoder, Nicole; Tal, Reshef; Martin, J Ryan

    2016-10-19

    Ectopic pregnancy is the leading cause of maternal morbidity and mortality during the first trimester and the incidence increases dramatically with assisted-reproductive technology (ART), occurring in approximately 1.5-2.1 % of patients undergoing in-vitro fertilization (IVF). Abdominal ectopic pregnancy is a rare yet clinically significant form of ectopic pregnancy due to potentially high maternal morbidity. While risk factors for ectopic pregnancy after IVF have been studied, very little is known about risk factors specific for abdominal ectopic pregnancy. We present a case of a 30 year-old woman who had an abdominal ectopic pregnancy following IVF and elective single embryo transfer, which was diagnosed and managed by laparoscopy. We performed a systematic literature search to identify case reports of abdominal or heterotopic abdominal ectopic pregnancies after IVF. A total of 28 cases were identified. Patients' ages ranged from 23 to 38 (Mean 33.2, S.D. = 3.2). Infertility causes included tubal factor (46 %), endometriosis (14 %), male factor (14 %), pelvic adhesive disease (7 %), structural/DES exposure (7 %), and unexplained infertility (14 %). A history of ectopic pregnancy was identified in 39 % of cases. A history of tubal surgery was identified in 50 % of cases, 32 % cases having had bilateral salpingectomy. Transfer of two embryos or more (79 %) and fresh embryo transfer (71 %) were reported in the majority of cases. Heterotopic abdominal pregnancy occurred in 46 % of cases while 54 % were abdominal ectopic pregnancies. Our systematic review has revealed several trends in reported cases of abdominal ectopic pregnancy after IVF including tubal factor infertility, history of tubal ectopic and tubal surgery, higher number of embryos transferred, and fresh embryo transfers. These are consistent with known risk factors for ectopic pregnancy following IVF. Further research focusing on more homogenous population may help in better characterizing

  15. Treatment period and medical care costs to achieve the first live birth by assisted reproductive technology are lower in the single embryo transfer period than in the double embryo transfer period: a retrospective analysis of women younger than 40 years of age.

    Science.gov (United States)

    Kawahara, Shunsuke; Ueda, Akane; Nakahori, Takashi; Honda, Tetsuro

    2017-04-01

    It was examined whether the single embryo transfer policy makes the treatment period longer for couples to achieve their first live birth by assisted reproductive technology. This study retrospectively analyzed women who started assisted reproductive technology at younger than 40 years of age in the authors' organization. The treatment periods for couples to achieve the first live birth by assisted reproductive technology, between the women who started assisted reproductive technology from 2004 to 2009 (the double embryo transfer period group, n=250), in which the double embryo transfer was predominant, and the women who started assisted reproductive technology from 2010 to 2015 (the single embryo transfer period group, n=298), in which the single embryo transfer was predominant, were compared. The age at the start of assisted reproductive technology, pregnancy rate per embryo transfer, and rate of women who achieved a live birth by assisted reproductive technology per number of women who tried assisted reproductive technology were all significantly higher in the single embryo transfer period group. Among the women who achieved a live birth by assisted reproductive technology, the incidence of multiple births and severe ovarian hyperstimulation syndrome, the treatment period, and medical care costs needed to achieve the first live birth were all significantly lower in the single embryo transfer period group. In the single embryo transfer period group, those women who were younger than 40 years of age achieved their first live birth by assisted reproductive technology more safely, quickly, and reasonably.

  16. The early-stage diagnosis of albinic embryos by applying optical coherence tomography

    Science.gov (United States)

    Yang, Bor-Wen; Wang, Shih-Yuan; Wang, Yu-Yen; Cai, Jyun-Jhang; Chang, Chung-Hao

    2013-09-01

    Albinism is a kind of congenital disease of abnormal metabolism. Poecilia reticulata (guppy fish) is chosen as the model to study the development of albinic embryos as it is albinic, ovoviviparous and with short life period. This study proposed an imaging method for penetrative embryo investigation using optical coherence tomography. By imaging through guppy mother’s reproduction purse, we found the embryo’s eyes were the early-developed albinism features. As human’s ocular albinism typically appear at about four weeks old, it is the time to determine if an embryo will grow into an albino.

  17. Comparison of reprogramming genes in induced pluripotent stem cells and nuclear transfer cloned embryos.

    Science.gov (United States)

    Duan, Lian; Wang, Zhendong; Shen, Jingling; Shan, Zhiyan; Shen, Xinghui; Wu, Yanshuang; Sun, Ruizhen; Li, Tong; Yuan, Rui; Zhao, Qiaoshi; Bai, Guangyu; Gu, Yanli; Jin, Lianhong; Lei, Lei

    2014-08-01

    The most effective reprogramming methods, somatic cell nuclear transfer (SCNT) and induced pluripotent stem cells (iPSCs), are widely used in biological research and regenerative medicine, yet the mechanism that reprograms somatic cells to totipotency remains unclear and thus reprogramming efficiency is still low. Microarray technology has been employed in analyzing the transcriptomes changes during iPS reprogramming. Unfortunately, it is difficult to obtain enough DNA from SCNT reconstructed embryos to take advantage of this technology. In this study, we aimed to identify critical genes from the transcriptional profile for iPS reprogramming and compared expression levels of these genes in SCNT reprogramming. By integrating gene expression information from microarray databases and published studies comparing somatic cells with either miPSCs or mouse embryonic stem cells (ESCs), we obtained two lists of co-upregulated genes. The gene ontology (GO) enriched analysis of these two lists demonstrated that the reprogramming process is associated with numerous biological processes. Specifically, we selected 32 genes related to heterochromatin, embryonic development, and cell cycle from our co-upregulated gene datasets and examined the gene expression level in iPSCs and SCNT embryos by qPCR. The results revealed that some reprogramming related genes in iPSCs were also expressed in SCNT reprogramming. We established the network of gene interactions that occur with genes differentially expressed in iPS and SCNT reprogramming and then performed GO analysis on the genes in the network. The network genes function in chromatin organization, heterochromatin, transcriptional regulation, and cell cycle. Further researches to improve reprogramming efficiency, especially in SCNT, will focus on functional studies of these selected genes.

  18. Association between ABO blood type and live-birth outcomes in single-embryo transfer cycles.

    Science.gov (United States)

    Pereira, Nigel; Patel, Hency H; Stone, Logan D; Christos, Paul J; Elias, Rony T; Spandorfer, Steven D; Rosenwaks, Zev

    2017-11-01

    To investigate the association between ABO blood type and live-birth outcomes in patients undergoing IVF with day 5 single-embryo transfer (SET). Retrospective cohort study. University-affiliated center. Normal responders, blood type and live birth, while controlling for confounders. Odds ratios (OR) with 95% confidence intervals (CI) for live birth were estimated. A total of 2,329 patients were included. The mean age of the study cohort was 34.6 ± 4.78 years. The distribution of blood types was as follows: A = 897 (38.5%); B = 397 (17.0%); AB = 120 (5.2%); and, O = 1,915 (39.3%) patients. There was no difference in the baseline demographics, ovarian stimulation, or embryo quality parameters between the blood types. The unadjusted ORs for live birth when comparing blood type A (referent) with blood types B, AB, and O were 0.96 (95% CI, 0.6-1.7), 0.72 (95% CI, 0.4-1.2), and 0.96 (95% CI. 0.6-1.7), respectively. The adjusted ORs for live birth remained not significant when comparing blood type A to blood types B, AB, and O individually. No difference in birth weight or gestational age at delivery was noted among the four blood types. Our findings suggest that ABO blood type is not associated with live-birth rate, birth weight, or gestational age at delivery in patients undergoing IVF with day 5 SET. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  19. Predictive value of plasma human chorionic gonadotropin measured 14 days after Day-2 single embryo transfer.

    Science.gov (United States)

    Løssl, Kristine; Oldenburg, Anna; Toftager, Mette; Bogstad, Jeanette; Praetorius, Lisbeth; Zedeler, Anne; Yding Andersen, Claus; Grøndahl, Marie L; Pinborg, Anja

    2017-08-01

    Prediction of pregnancy outcome after in vitro fertilization is important for patients and clinicians. Early plasma human chorionic gonadotropin (p-hCG) levels are the best known predictor of pregnancy outcome, but no studies have been restricted to single embryo transfer (SET) of Day-2 embryos. The aim of the present study was to investigate the predictive value of p-hCG measured exactly 14 days after the most commonly used Day-2 SET on pregnancy, delivery, and perinatal outcome. A retrospective analysis of prospectively collected data on 466 women who had p-hCG measured exactly 14 days after Day-2 SET during a randomized trial including 1050 unselected women (aged 18-40 years) undergoing their first in vitro fertilization/ intracytoplasmic sperm injection treatment. The p-hCG predicted clinical pregnancy [area under the curve (AUC) 0.953; 95% CI 0.915-0.992] significantly better than ongoing pregnancy (AUC 0.803, 95% CI 0.717-0.890) and delivery (AUC 0.772, 95% CI 0.691-0.854). Women with p-hCG levels in the lowest quartile had significantly lower clinical pregnancy, ongoing pregnancy, and delivery rates (p < 0.001), whereas the pregnancy outcome and post-clinical pregnancy loss remained similar throughout the three highest p-hCG quartiles. The p-hCG level was related to neither birthweight nor gestational age at delivery. Clinical pregnancy is significantly better predicted by p-hCG compared with ongoing pregnancy and delivery. Clinical pregnancy rates, ongoing pregnancy rates, and delivery rates remained similar throughout the three highest p-hCG quartiles with no trend towards "the higher the better". © 2017 Nordic Federation of Societies of Obstetrics and Gynecology.

  20. Differential expression of metallothionein isoforms in terrestrial snail embryos reflects early life stage adaptation to metal stress.

    Science.gov (United States)

    Baurand, Pierre-Emmanuel; Pedrini-Martha, Veronika; de Vaufleury, Annette; Niederwanger, Michael; Capelli, Nicolas; Scheifler, Renaud; Dallinger, Reinhard

    2015-01-01

    The aim of this study was to analyze the expression of three metallothionein (MT) isoform genes (CdMT, CuMT and Cd/CuMT), already known from adults, in the Early Life Stage (ELS) of Cantareus aspersus. This was accomplished by detection of the MT isoform-specific transcription adopting Polymerase Chain Reaction (PCR) amplification and quantitative Real Time (qRT)-PCR of the three MT genes. Freshly laid eggs were kept for 24 hours under control conditions or exposed to three cadmium (Cd) solutions of increasing concentration (5, 10, and 15 mg Cd/L). The transcription of the three MT isoform genes was detected via PCR in 1, 6 and 12-day-old control or Cd-exposed embryos. Moreover, the transcription of this isoform genes during development was followed by qRT-PCR in 6 and 12-day-old embryos. Our results showed that the CdMT and Cd/CuMT genes, but not the CuMT gene, are expressed in embryos at the first day of development. The transcription of the 3 MT genes in control embryos increased with development time, suggesting that the capacities of metal regulation and detoxification may have gradually increased throughout embryogenesis. However in control embryos, the most highly expressed MT gene was that of the Cd/CuMT isoform, whose transcription levels greatly exceeded those of the other two MT genes. This contrasts with the minor significance of this gene in adult snails and suggests that in embryos, this isoform may play a comparatively more important role in metal physiology compared to adult individuals. This function in adult snails appears not to be related to Cd detoxification. Instead, snail embryos responded to Cd exposure by over-expression of the CdMT gene in a concentration-dependent manner, whereas the expression of the Cd/CuMT gene remained unaffected. Moreover, our study demonstrates the ability of snail embryos to respond very early to Cd exposure by up-regulation of the CdMT gene.

  1. Application of seminal plasma to female genital tract prior to embryo transfer in assisted reproductive technology cycles (IVF, ICSI and frozen embryo transfer).

    Science.gov (United States)

    Ata, Baris; Abou-Setta, Ahmed M; Seyhan, Ayse; Buckett, William

    2018-02-28

    The female genital tract is not exposed to seminal plasma during standard assisted reproductive technology (ART) cycles. However, it is thought that the inflammatory reaction triggered by seminal plasma may be beneficial by inducing maternal tolerance to paternal antigens expressed by the products of conception, and may increase the chance of successful implantation and live birth. To assess the effectiveness and safety of application of seminal plasma to the female genital tract prior to embryo transfer in ART cycles. We searched the following databases from inception to October 2017: Cochrane Gynaecology and Fertility Group Specialised Register of Controlled Trials, Cochrane Central Register of Studies Online (CRSO), MEDLINE, Embase, CINAHL and PsycINFO. We also searched trial registers for ongoing trials, including International Clinical Trials Registry Platform (ICTRP) Search Portal and ClinicalTrials.gov. Other sources searched were; Web of Knowledge, OpenGrey, LILACS, PubMed, Google Scholar and the reference lists of relevant articles. We included randomised controlled trials (RCTs) conducted among women undergoing ART, comparing any procedure that would expose the female genital tract to seminal plasma during the period starting five days before embryo transfer and ending two days after it versus no seminal plasma application. Two review authors independently selected trials, assessed risk of bias, and extracted data. We pooled data to calculate relative risks (RRs) and 95% confidence intervals (CIs). We assessed statistical heterogeneity using the I 2 statistic. We assessed the overall quality of the evidence for the main outcomes using GRADE methods. Our primary outcomes were live birth rate and miscarriage rate. Secondary outcomes were live birth/ongoing pregnancy rate, clinical pregnancy rate, multiple pregnancy rate, ectopic pregnancy rate and the incidence of other adverse events. We included 11 RCTs (3215 women). The quality of the evidence ranged

  2. A randomized double-blinded controlled trial of hCG as luteal phase support in natural cycle frozen embryo transfer.

    Science.gov (United States)

    Lee, Vivian Chi Yan; Li, Raymond Hang Wun; Yeung, William Shu Biu; Pak Chung, H O; Ng, Ernest Hung Yu

    2017-05-01

    Does the use of hCG as luteal phase support in natural cycle frozen embryo transfer (FET) increase the ongoing pregnancy rate? The use of hCG in natural cycle FET did not improve the ongoing pregnancy rate. The use of luteal phase support in stimulated cycles has been associated with higher live-birth rates and the results are similar when using hCG or progesterone. This is a randomized double-blinded controlled trial of 450 women recruited between August 2013 and October 2015. Women with regular cycles undergoing natural cycle FET were recruited. Serial serum hormonal concentrations were used to time natural ovulation and at least Day 2 cleavage embryos were replaced. Patients were randomized into either: (i) the treatment group, receiving 1500 IU hCG on the day of FET and 6 days after FET, or (ii) the control group, receiving normal saline on these 2 days. The ongoing pregnancy rate [60/225 (26.7%) in the treatment group vs 70/225 (31.3%) in the control group, odds ratio 1.242 (95% CI 0.825-1.869)], implantation rate and miscarriage rate were comparable between the two groups. In the treatment group, there were significantly more cycles with top quality embryos transferred and a significantly higher serum oestradiol level, but a comparable serum progesterone level, 6 days after FET. However, no significant differences were observed in serum oestradiol and progesterone levels 6 days after FET between the pregnant and non-pregnant women. In the multivariate logistic regression, the number of embryos transferred was the only significant factor predictive of the ongoing pregnancy rate after natural cycle FET. This study only included FET with cleavage stage embryos and only hCG, not vaginal progesterone, was used as luteal phase support. The findings in this study do not support the use of hCG for luteal phase support in natural cycle FET. No external funding was used and there were no competing interests. clinicaltrial.gov identifier: NCT01931384. 23/8/2013. 30

  3. The Effect of Cirata Reservoir Sediment on Early Developmental Stage of Common Carp (Cyprinus carpio Embryo

    Directory of Open Access Journals (Sweden)

    Yuni Pujihastuti

    2009-07-01

    Full Text Available Sedimentation at Cirata reservoir may directly and indirectly influence fish particularly fish which have an adhesive characteristic at its early developmental stage such as common carp (Cyprinus carpio. Sample of sediment was collected from Cirata reservoir using Eikmand dredge at a depth of 80 m. The sample was subsequently centrifuged at 5500 rpm for 10 min. The supernatant obtained was then used for toxicity test on common carp at early developmental stage. In this test, four treatments were applied based on the concentration of sediment supernatant, namely: 0, 8.33, 16.60 and 24.90 %. The results showed that a higher sediment supernatant concentration resulted in lower egg yolk absorption rate, lower relative growth rate in length, lower egg yolk efficiency and higher egg and larval abnormality.  Higher sediment supernatant concentration also resulted in lower hatching percentage of common carp larva. The damage of eggs and larval morphologies in treatments with sediment supernatant was likely caused by the presence Pb and organic matters which act in synergy. Keywords :  sediment, Cirata, embryo, common carp   ABSTRAK Sedimentasi di Waduk Cirata secara langsung dan tidak langsung akan berpengaruh terhadap kehidupan ikan khususnya tahap awal perkembangan ikan yang bersifat adhesiveseperti ikan mas (Cyprinus carpio.  Sampel sedimen waduk Cirata diambil dengan Eikmand dredge pada kedalaman 80 m.  Hasil ekstrak di sentrifugasi dengan kecepatan 5500 rpm selama 10 menit untuk diambil air pori sedimennya.  Air pori digunakan sebagai bahan uji toksisitas terhadap perkembangan awal ikan mas dengan perlakuan 0; 8,33; 16,60 dan 24,90 %. Hasil uji toksisitas diperoleh bahwa semakin tinggi konsentrasi air pori dari sediment maka semakin rendah laju penyerapan kuning telur Laju pertumbuhan relatif panjang embrio pada berbagai konsentrasi juga diperoleh bahwa semakin tinggi konsentrasi air sedimen maka semakin rendah laju pertumbuhan relatif

  4. Characterization of the altered gene expression profile in early porcine embryos generated from parthenogenesis and somatic cell chromatin transfer.

    Science.gov (United States)

    Zhou, Chi; Dobrinsky, John; Tsoi, Stephen; Foxcroft, George R; Dixon, Walter T; Stothard, Paul; Verstegen, John; Dyck, Michael K

    2014-01-01

    The in vitro production of early porcine embryos is of particular scientific and economic interest. In general, embryos produced from in vitro Assisted Reproductive Technologies (ART) manipulations, such as somatic cell chromatin transfer (CT) and parthenogenetic activation (PA), are less developmentally competent than in vivo-derived embryos. The mechanisms underlying the deficiencies of embryos generated from PA and CT have not been completely understood. To characterize the altered genes and gene networks in embryos generated from CT and PA, comparative transcriptomic analyses of in vivo (IVV) expanded blastocysts (XB), IVV hatched blastocyst (HB), PA XB, PA HB, and CT HB were performed using a custom microarray platform enriched for genes expressed during early embryonic development. Differential expressions of 1492 and 103 genes were identified in PA and CT HB, respectively, in comparison with IVV HB. The "eIF2 signalling", "mitochondrial dysfunction", "regulation of eIF4 and p70S6K signalling", "protein ubiquitination", and "mTOR signalling" pathways were down-regulated in PA HB. Dysregulation of notch signalling-associated genes were observed in both PA and CT HB. TP53 was predicted to be activated in both PA and CT HB, as 136 and 23 regulation targets of TP53 showed significant differential expression in PA and CT HB, respectively, in comparison with IVV HB. In addition, dysregulations of several critical pluripotency, trophoblast development, and implantation-associated genes (NANOG, GATA2, KRT8, LGMN, and DPP4) were observed in PA HB during the blastocyst hatching process. The critical genes that were observed to be dysregulated in CT and PA embryos could be indicative of underlying developmental deficiencies of embryos produced from these technologies.

  5. Day-3 Medium Changes can Affect Developmental Potential of Porcine Somatic Cell Nuclear Transfer and Parthenogenesis Embryos In Vitro

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    Dibyendu Biswas and Sang Hwan Hyun*

    2011-01-01

    Full Text Available The aim of the present study was to compare the developmental competence of porcine parthenotes and somatic cell nuclear transfer (SCNT embryos after day-3 medium change with fresh embryo culture medium to that of embryos that did not have a medium change (monoculture system. The parthenogenetic and SCNT blastocyst formation rates were significantly (P<0.05 higher in the no-medium-change group (43.3±2.3, 18.5±1.1%, respectively compared with the day-3 medium-change group (35.9±2.4, 7.9±0.9%, respectively. Total cell number in parthenotes and SCNT blastocysts was also significantly (P<0.05 higher in the no-media-change group (92.0±4.2, 66.9±7.7, respectively compared with the media-change group (81.5±3.1, 46.6±4.9, respectively. No significant difference in cleavage rate was found in either group for parthenotes or SCNT embryos. This result suggests that day-3 medium changes have negative effects on porcine parthenotes and SCNT embryos in vitro.

  6. Vitrification in Open and Closed Carriers at Different Cell Stages: Assessment of Embryo Survival, Development, DNA Integrity and Stability during Vapor Phase Storage for Transport

    Directory of Open Access Journals (Sweden)

    Goldberg Jeffrey

    2011-03-01

    Full Text Available Abstract Background High cooling rates with vitrification can be achieved through the use of carriers that allow cryopreservation in fluid volumes Methods Frozen one-cell mouse embryos were thawed and randomly allocated to treatment groups. Embryos were cultured and vitrified at the 8-cell (CL or at the blastocyst (BL stage. The cryoloop, an open carrier was tested against two closed systems, the Cryotip and the HSV straw. Carriers were tested for their ability to maintain embryo viability when held in the vapor phase of a dry shipper for a period of 96 hours. Outcome parameters monitored were embryo survival, recovery, subsequent development and signs of DNA damage. Results A total of 561 embryos were vitrified. The only parameter significantly affected by the type of carrier was the percentage of embryos recovered after warming. Vitrification of both CL and BL stage embryos in the Cryotip resulted in significantly lower recovery rates (P Conclusion This study is one of the first to examine DNA integrity after vitrification on different carriers and at different cell stages. It also provides insight on relative safety of short term vapor storage of vitrified embryos during transport. Within the limits of this study we could not detect an adverse effect of vapor storage on blastomere DNA or other measured outcome parameters.

  7. Identification and quantitative analysis of stage-specific carbohydrates in loblolly pine (Pinus taeda) zygotic embryo and female gametophyte tissues.

    Science.gov (United States)

    Pullman, Gerald S; Buchanan, Mike

    2008-07-01

    Stage-specific analyses of starch and 18 sugars, including pentoses, hexoses, disaccharides, trisaccharides, oligosaccharides and sugar alcohols, were made throughout seed development for zygotic embryo and female gametophyte (FG) tissues of loblolly pine (Pinus taeda L.). Tissue was most often analyzed in triplicate from two open-pollinated families grown in different locations and sampled in different years. Carbohydrates were analyzed by enzymatic assay, high performance liquid chromatography or gas chromatography/mass spectrometry. For all carbohydrates quantified, peak concentrations were higher in embryo tissue than in FG tissue. Significant changes in starch and sugar concentrations occurred over time, with both seed collections showing similar trends in temporal changes. Although concentrations were not always similar, embryo and FG tissues generally showed similar patterns of change in starch and sugar concentrations over time. Total starch concentration was highest during early seed development and decreased as development progressed. The major sugars contributing to osmotic potential during early seed development were D-pinitol, sucrose, fructose and glucose. During mid-seed development, D-pinitol, sucrose, fructose, glucose, melibiose and raffinose provided major contributions to the osmotic environment. During late seed development, sucrose, raffinose, melibiose, stachyose and fructose were the major contributors to osmotic potential. These data suggest stage-specific media composition for each step in the somatic embryogenesis protocol.

  8. Intravenous intralipid therapy is not beneficial in having a live delivery in women aged 40-42 years with a previous history of miscarriage or failure to conceive despite embryo transfer undergoing in vitro fertilization-embryo transfer.

    Science.gov (United States)

    Check, J H; Check, D L

    2016-01-01

    To evaluate the efficacy of intralipid intravenous infusion in achieving a live pregnancy following IVF--embryo transfer in women of advanced reproductive age (40-42 years). A matched control was performed. Women aged 40-42 with a previous history of miscarriage or who failed to conceive despite previous embryo transfer who entered an IVF program were offered intravenous intralipid therapy (four ml of 20% liposyn II in 100 ml normal saline over one hour) during the mid-follicular phase. Clinical pregnancy rates (eight weeks with viable gestation) and live delivered pregnancy rates were then determined and compared. The results were evaluated after ten matched cycles. There were no clinical pregnancies in those receiving intralipid vs. a 40% clinical and a 30% live delivered pregnancy rate in the untreated controls (p = 0.087, Fisher's exact test). The study was terminated because of these preliminary data. In the test tube, adding intralipid to natural killer cells can inhibit their cytolytic action. However, the use of intravenous intralipid to suppress natural killer cell activity does not seem to improve the chance of a live delivery in women aged 40-42 years with a previous history of miscarriage. In fact this therapy may actually be detrimental in this age group. Since efficacy of this therapy was not found in a group of advanced reproductive age, it is not clear why this should be effective for a younger population. A controlled study for the younger group is needed. Perhaps such a study could be limited to only those with miscarriage rather than also concluding failure to conceive despite embryo transfer. Intralipid failed to improve live delivered pregnancy rates in women with prior miscarriage or previous failure with embryo transfer.

  9. Changes in the transcriptome of morula-stage bovine embryos caused by heat shock: relationship to developmental acquisition of thermotolerance

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    Sakatani Miki

    2013-01-01

    Full Text Available Abstract Background While initially sensitive to heat shock, the bovine embryo gains thermal resistance as it progresses through development so that physiological heat shock has little effect on development to the blastocyst stage by Day 5 after insemination. Here, experiments using 3’ tag digital gene expression (3’DGE and real-time PCR were conducted to determine changes in the transcriptome of morula-stage bovine embryos in response to heat shock (40 degrees C for 8 h that could be associated with thermotolerance. Results Using 3’DGE, expression of 173 genes were modified by heat shock, with 94 genes upregulated by heat shock and 79 genes downregulated by heat shock. A total of 38 differentially-regulated genes were associated with the ubiquitin protein, UBC. Heat shock increased expression of one heat shock protein gene, HSPB11, and one heat shock protein binding protein, HSPBP1, tended to increase expression of HSPA1A and HSPB1, but did not affect expression of 64 other genes encoding heat shock proteins, heat shock transcription factors or proteins interacting with heat shock proteins. Moreover, heat shock increased expression of five genes associated with oxidative stress (AKR7A2, CBR1, GGH, GSTA4, and MAP2K5, decreased expression of HIF3A, but did not affect expression of 42 other genes related to free radical metabolism. Heat shock also had little effect on genes involved in embryonic development. Effects of heat shock for 2, 4 and 8 h on selected heat shock protein and antioxidant genes were also evaluated by real-time PCR. Heat shock increased steady-state amounts of mRNA for HSPA1A (PHSP90AA1 (PSOD1 or CAT. Conclusions Changes in the transcriptome of the heat-shocked bovine morula indicate that the embryo is largely resistant to effects of heat shock. As a result, transcription of genes involved in thermal protection is muted and there is little disruption of gene networks involved in embryonic development. It is likely that

  10. The Importance of the Periconception Period: Immediate Effects in Cattle Breeding and in Assisted Reproduction Such as Artificial Insemination and Embryo Transfer.

    Science.gov (United States)

    Van Eetvelde, Mieke; Heras, Sonia; Leroy, J L M R; Van Soom, Ann; Opsomer, Geert

    2017-01-01

    In livestock breeding, the successful outcome is largely depending on the "periconception environment" which, in a narrow sense, refers to the genital tract, where gametogenesis and embryogenesis occur. During these early stages of development, gametes and embryos are known to be particularly sensitive to alterations in their microenvironment. However, as the microenvironment somehow reflects what is going on in the external world, we must widen our definition of "periconception environment" and refer to all events taking place around the time of conception, including metabolic state and health and nutrition of the dam. In modern dairy cows that have to manage an optimal reproductive performance with continued growth and high milk yield, the periconception period is particularly challenging. The metabolic priority for growth and lactation is known to generate adverse conditions hampering optimal ovarian function, oocyte maturation, and development of embryo/fetus. In addition, by using artificial reproductive technologies (ARTs), gametes and/or embryos of livestock are exposed to unnatural conditions outside the male and female genital tract. Artificial insemination, the most widely used technique, is currently yielding pregnancy rates similar to natural mating, and calves produced by AI are equally viable after natural mating. In contrast, other ART, such as multiple ovulation and embryo transfer, have been reported to induce changes in gene expression and DNA methylation patterns with potential consequences for development.Finally, the "periconceptional" environment has been shown to not only influence the successful establishment of pregnancy but also the long-term health and productivity of the offspring. Hence, the optimization of management around the time of conception might open doors to improve animal production and product quality.

  11. Embryo transfers between C57BL/6J and DBA/2J mice: Examination of a maternal effect on ethanol teratogenesis

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    David eGilliam

    2014-12-01

    Full Text Available Genetic factors influence Fetal Alcohol Spectrum Disorders (FASD in both humans and animals. Experiments using inbred and selectively bred mouse stocks that controlled for 1 ethanol dose, 2 maternal and fetal blood ethanol levels, and 3 fetal developmental exposure stage, show genotype can affect teratogenic outcome. Other experiments distinguish the teratogenic effects mediated by maternal genotype from those mediated by fetal genotype. One technique to distinguish maternal versus fetal genotype effect is to utilize embryo transfers. This study is the first to examine ethanol teratogenesis - fetal weight deficits and mortality, and digit, kidney, and vertebral malformations - in C57BL/6J (B6 and DBA/2J (D2 fetuses that were transferred as blastocysts into B6 and D2 dams. We hypothesized that, following maternal alcohol exposure, B6 and D2 fetuses gestating within B6 mothers, as compared to D2 mothers, will exhibit a higher frequency of malformations. On day 9 of pregnancy, females were intubated (IG with either 5.8 g/kg ethanol (E or maltose dextrin (MD. Other females were mated within strain and treated with either ethanol or maltose, or were not exposed to either treatment. Implantation rates were affected by genotype. Results show more B6 embryos implanted into D2 females than B6 females (p<.05; 47% vs 23%, respectively. There was no difference in the percentage of D2 embryos implanting into B6 and D2 females (14% and 16%, respectfully. Litter mortality averaged 24% across all experimental groups. Overall, in utero ethanol exposure reduced mean litter weight compared to maltose treatment (E=1.01 g; MD=1.19 g; p<.05; but maltose exposed litters with transferred embryos weighed more than similarly treated natural litters (1.30 g vs 1.11 g; p<.05. Approximately 50% of all ethanol exposed B6 fetuses exhibited some malformation (digit, vertebral, and/or kidney regardless of whether they were transferred into a B6 or D2 female, or were naturally

  12. Mesenteric vein thrombosis following impregnation via in vitro fertilization-embryo transfer.

    Science.gov (United States)

    Hirata, Masaaki; Yano, Hiroko; Taji, Tomoe; Shirakata, Yoshiharu

    2017-10-27

    Pregnancy is an acquired hypercoagulable state. Most patients with thrombosis that develops during pregnancy present with deep vein leg thrombosis and/or pulmonary embolism, whereas the development of mesenteric vein thrombosis (MVT) in pregnant patients is rare. We report a case of MVT in a 34-year-old woman who had achieved pregnancy via in vitro fertilization-embryo transfer (IVF-ET). At 7 wk of gestation, the patient was referred to us due to abdominal pain accompanied by vomiting and hematochezia, and she was diagnosed with superior MVT. Following resection of the gangrenous portion of the small intestine, anticoagulation therapy with unfractionated heparin and thrombolysis therapy via a catheter placed in the superior mesenteric artery were performed, and the patient underwent an artificial abortion. Oral estrogen had been administered for hormone replacement as part of the IVF-ET procedure, and additional precipitating factors related to thrombosis were not found. Pregnancy itself, in addition to the administered estrogen, may have caused MVT in this case. We believe that MVT should be included in the differential diagnosis of a pregnant patient who presents with an acute abdomen.

  13. Heterotopic pregnancy after in vitro fertilization and embryo transfer: the role of routine transvaginal ultrasonography

    International Nuclear Information System (INIS)

    Lee, Eun Ju

    1994-01-01

    Spontaneous heterotopic pregnancy (HP) is a rare condition, however, the rising incidence after in-vitro fertilization and embryo transfer (IVF-ET) may present serious problems as the diagnosis of this potentially fatal condition is often missed. The purposes of this study were to investigate the frequency of HP after IVF-ET, to analyze the clinical presentation and the outcome of coincidental intrauterine pregnancy, and to evaluate the value of routine transvaginal ultrasonography in early diagnosis and treatment of HP. There were 11 cases of HP among 484 pregnancies after IVF-ET (The incidence was 2.3%) in recent 2 years. We evaluated the clinical findings and the findings of transvaginal sonography in those 11 patients. The combined extrauterine pregnancies were tubal (n = 9), cornual (n = 1), and both the tubal and cornual (n = 1). Vaginal bleeding was absent in 4 cases and one of them was asymptomatic. HP may be suspected if there was an elevated serum hCG in the absence of intrauterine multiple gestations. In all cases, the condition was diagnosed by transvaginal sonography performed prior to surgical intervention. The coincidental intrauterine pregnancies were ended in successful full-term delivery in 7 cases, while one pregnancy was ongoing at that time. The incidence of HP was evidently higher in IVF-ET than in spontaneous pregnancies. Transvaginal sonography was useful in accurate diagnosis in clinically suspected cases and even in asymptomatic cases. Routine transvaginal sonography played an important role in early diagnosis and prompt management of HP

  14. Effect of meloxicam on pregnancy rate of recipient heifers following transfer of in vitro produced embryos.

    Science.gov (United States)

    Aguiar, T S; Araújo, C V; Tirloni, R R; Martins, L R

    2013-12-01

    The main objective of this study was to determine if administration of meloxicam, a cyclooxygenase (COX) two inhibitor, to heifers in which embryo transfer (ET) is more difficult and requires a greater manipulation of the tract, would be beneficial. Nulliparous recipient heifers were divided in two groups: CON (n = 102), in which animals received 10 ml of saline IM (the same volume of meloxicam) and MEL (n = 105) animals that were treated with meloxicam. According to the degree in passing the catheter, recipients from both groups were classified as Grade I, easy (meloxicam (10 ml).There was no difference in the pregnancy rates on Day 35 considering animals which presented Grade I cervix independently whether the treatment was performed or not (p = 0.22). There was a statistical difference in the pregnancy rates (p animals that presented Grade II cervix, the pregnancy rate was higher for MEL (21.15% and 78.84%, respectively) in both examinations (p meloxicam had a positive influence on general pregnancy rate of treated heifers in comparison to non-treated heifers. It was also observed that pregnancy rate was not influenced by meloxicam administration in Grade I heifers. Treatment increased the pregnancy rate of Grade II heifers. © 2013 Blackwell Verlag GmbH.

  15. [Retrospective analysis of 6 cases of miliary tuberculosis in pregnant women after in vitro fertilization and embryo transfer].

    Science.gov (United States)

    Gao, Hui-juan; Hu, Yan-jun; Zhu, Yi-min; Huang, He-feng

    2007-11-01

    To analyze the clinical characteristics of haematogenous disseminated tuberculosis in pregnant women after in vitro fertilization and embryo transfer (IVF-ET). Six cases of haematogenous disseminated tuberculosis after in vitro fertilization pregnancy from January 2001 to December 2005 were retrospectively analyzed. The clinical manifestations of pulmonary miliary tuberculosis were non-specific. All cases presented with fever for several weeks, but typical respiratory manifestations were lacking. Chest X-ray showed typical miliary and infiltrate lesions. The onset of fever was at an average of 53.2 days after embryo transfer. Miscarriage occurred in 5 patients within 2 to 4 weeks after the onset of fever. The main symptom of miliary tuberculosis in pregnant women after IVF-ET was fever, and the prognosis of the pregnancy was poor. Physician should be alert to the occurrence of haematogenous disseminated tuberculosis, especially in pregnant women with long-term fever.

  16. The efficacy of intrauterine injection of human chorionic gonadotropin before embryo transfer in assisted reproductive cycles: Meta-analysis.

    Science.gov (United States)

    Ye, Hong; Hu, Jing; He, Wencong; Zhang, Yong; Li, Caihong

    2015-12-01

    A systematic review and meta-analysis to evaluate the effect of human chorionic gonadotropin (hCG) intrauterine injection before embryo transfer on the outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Searches of PubMed®, EMBASE®, EBSCO, Web of Science®, SCOPUS® and Cochrane Central Register of Controlled Trials were conducted to retrieve relevant randomized controlled trials (RCTs). Data were extracted and analysed. The meta-analysis included five RCTs (hCG group n = 680; control group n = 707). Intrauterine hCG injection significantly increased rates of biochemical, clinical and ongoing pregnancy compared with controls. There were no between-group differences in implantation or miscarriage rates. Women undergoing IVF/ICSI may benefit from intrauterine hCG injection before embryo transfer. © The Author(s) 2015.

  17. Amniotic fluid l-ergothioneine concentrations in pregnant sheep after natural mating and transfer of vitrified/thawed in-vitro produced embryos.

    Science.gov (United States)

    Sotgia, Salvatore; Zinellu, Angelo; Arru, Dionigia; Nieddu, Stefano; Strina, Alessandro; Ariu, Federica; Pintus, Gianfranco; Carru, Ciriaco; Bogliolo, Luisa; Ledda, Sergio

    2015-10-01

    L-ergothioneine levels were measured in amniotic fluid of pregnant sheep after natural mating and transfer of vitrified/thawed in-vitro produced embryos. Amniotic fluids were collected between 60 and 65 and 80-85 days of gestation and analysed by an ultra-performance liquid chromatographic (UPLC)method with fluorescence detection. L-Ergothioneine concentrations ranged between 0.23 and 9.36 μmol/L and were significantly higher in pregnancy obtained by the transfer of vitrified/thawed in-vitro produced embryos. Conversely, no significant changes in amniotic fluid L-ergothioneine concentrations were observed according to the stages of pregnancy considered in this study. These findings suggest that L-ergothioneine concentrations, are not affected as much by the gestational age, but rather by the method used to induce the pregnancy. On the whole, the measurement of L-ergothioneine in amniotic fluid could serve as a useful biomarker of oxidative stress and/or inflammatory state in pregnancy.

  18. Embryo wastage rates remain high in assisted reproductive technology (ART): a look at the trends from 2004-2013 in the USA.

    Science.gov (United States)

    Ghazal, Sanaz; Patrizio, Pasquale

    2017-02-01

    This work examined the trend in "embryo wastage" rates after ART in USA and its relationship to the number of embryos transferred, live born infants delivered across patient age, and the yearly percentage of embryos wasted. The data were obtained from the US-clinics SART databank for the years 2004-2013. A total of 1,808,082 non-donor embryos were transferred in 748,394 fresh cycles resulting in 358,214 liveborn. During the years of analysis, the mean number of embryos transferred has progressively decreased leading to an overall significant decrease in Embryo Wastage rates (83.2 to 76.5%, p ART cycle are competent. The overall "Embryo Wastage" rates have consistently decreased from a high of 90% in 1995 to a rate of 76.5% in 2013. Transferring fewer embryos particularly at the blastocyst-stage and improved methods of embryo selection may further decrease "Embryo Wastage" rates.

  19. Elective single-embryo transfer improves cumulative pregnancy outcome in young patients but not in women of advanced reproductive age.

    Science.gov (United States)

    Fujimoto, Akihisa; Morishima, Kaoru; Harada, Miyuki; Hirata, Tetsuya; Osuga, Yutaka; Fujii, Tomoyuki

    2015-12-01

    The purpose of the present study is to assess the significance of elective single-embryo transfer (eSET) in older women. The outcomes of assisted reproductive technology between 2001 and 2013 at single institution were retrospectively evaluated. Cumulative live birth rates (CLBRs) in one oocyte retrieval cycle were compared between those who underwent eSET and multiple embryo transfer (MET) in fresh cycles. The outcomes of 429 eSET cycles and 965 MET cycles were compared. CLBRs in eSET were higher than those of MET in women under 37 and were comparable in women aged 37 and over. The analysis of the outcomes separately in three age subgroups showed a significantly higher CLBR in young eSET (aged under 37) than that in young MET and similar CLBR between older (aged 37-40 and over 40) eSET and MET. Multiple birth rates were lower in eSET in all age groups. Multivariate logistic regression analyses showed that, in women aged under 37, number of frozen embryos, presence of good-quality embryos, and eSET were significantly related to cumulative live birth. In women aged between 37 and 40, age and number of frozen embryos were significantly related, while eSET was not. eSET in women under 37 resulted in increased CLBR compared with MET. In women aged between 37 and 40, CLBR in eSET group was similar with that in MET group. In both age groups, eSET reduced multiple birth rates. The significance of eSET in older women is limited presently, and further research on the strategy to improve cumulative outcomes is necessary.

  20. Cumulative results including obstetrical and neonatal outcome of fresh and frozen-thawed cycles in elective single versus double fresh embryo transfers.

    Science.gov (United States)

    Fauque, Patricia; Jouannet, Pierre; Davy, Céline; Guibert, Juliette; Viallon, Vivian; Epelboin, Sylvie; Kunstmann, Jean-Marie; Patrat, Catherine

    2010-08-01

    To assess the efficacy of elective single embryo transfer (e-SET) compared to a double embryo transfer (DET) in a selected population including obstetrical and neonatal outcome of fresh and frozen-thawed cycles. Prospective nonrandomized study. Department of reproductive medicine. Elective single embryo transfer was proposed to women 0.05). Twin delivery rate was significantly different between the two groups (3.5% versus 37.5% respectively, P<0.05). Neonatal outcome in twins resulting from IVF-ICSI was found to be poorer than in singletons, considering the mean gestational age, mode of delivery, birthweight, and risk of neonatal intensive care unit admission for the infants. In a selected population, the elective transfer of one embryo with high implantation potential helped to avoid twin pregnancies without decreasing delivery rate. Copyright (c) 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  1. Single stage incubators and Hypercapnia during incubation affect the vascularization of the chorioallantoic membrane in broiler embryos.

    Science.gov (United States)

    Fernandes, J I M; Bortoluzzi, C; Schmidt, J M; Scapini, L B; Santos, T C; Murakami, A E

    2017-01-01

    Incubation management can have direct effects on neonate health and consequently affect post-hatching development. The effects of incubation in multiple and single stage incubators with different concentrations of CO 2 were evaluated in terms of the vessel density in the chorioallantoic membrane, hatching, heart morphology, and body development of the neonate up to the tenth day. A total of 2,520 fertile eggs were used and distributed in a completely randomized design with 4 levels of CO 2 in 4 single-stage incubators (4,000; 6,000; 8,000; and 10,000 ppm) and a control treatment based on multiple-stage incubation, totaling 5 treatments. The levels of CO 2 were used during the first 10 d of the incubation period, and after this period, all eggs were submitted to the same level of CO 2 (4,000 ppm). Eggs that were incubated in multiple-stage incubators presented a lower percentage of vessels in the chorioallantoic membrane, lower yolk absorption by the embryo, wall depth of the right ventricle, and greater humidity losses in the eggs when compared to eggs in the single-stage incubators. The eggs submitted to hypercapnia, between 5,000 and 6,000 ppm of CO 2 , had a higher percentage of vessels in the chorioallantoic membrane; the embryos originating from these eggs had higher weight, with higher relative weight of the liver. However, the same levels reduced the yolk absorption. Single-stage incubation with moderate levels of hypercapnia is an efficient tool to be adopted by the hatcheries when attempting to improve chick quality. © 2016 Poultry Science Association Inc.

  2. Transfer of Dicamba Tolerance from Sinapis arvensis to Brassica napus via Embryo Rescue and Recurrent Backcross Breeding

    Science.gov (United States)

    Jugulam, M.; Ziauddin, Asma; So, Kenny K. Y.; Chen, Shu; Hall, J. Christopher

    2015-01-01

    Auxinic herbicides (e.g. dicamba) are extensively used in agriculture to selectively control broadleaf weeds. Although cultivated species of Brassicaceae (e.g. Canola) are susceptible to auxinic herbicides, some biotypes of Sinapis arvensis (wild mustard) were found dicamba resistant in Canada. In this research, dicamba tolerance from wild mustard was introgressed into canola through embryo rescue followed by conventional breeding. Intergeneric hybrids between S. arvensis (2n = 18) and B. napus (2n = 38) were produced through embryo rescue. Embryo formation and hybrid plant regeneration was achieved. Transfer of dicamba tolerance from S. arvensis into the hybrid plants was determined by molecular analysis and at the whole plant level. Dicamba tolerance was introgressed into B. napus by backcrossing for seven generations. Homozygous dicamba-tolerant B. napus lines were identified. The ploidy of the hybrid progeny was assessed by flow cytometry. Finally, introgression of the piece of DNA possibly containing the dicamba tolerance gene into B. napus was confirmed using florescence in situ hybridization (FISH). This research demonstrates for the first time stable introgression of dicamba tolerance from S. arvensis into B. napus via in vitro embryo rescue followed by repeated backcross breeding. Creation of dicamba-tolerant B. napus varieties by this approach may have potential to provide options to growers to choose a desirable herbicide-tolerant technology. Furthermore, adoption of such technology facilitates effective weed control, less tillage, and possibly minimize evolution of herbicide resistant weeds. PMID:26536372

  3. Accounting for unknown foster dams in the genetic evaluation of embryo transfer progeny.

    Science.gov (United States)

    Suárez, M J; Munilla, S; Cantet, R J C

    2015-02-01

    Animals born by embryo transfer (ET) are usually not included in the genetic evaluation of beef cattle for preweaning growth if the recipient dam is unknown. This is primarily to avoid potential bias in the estimation of the unknown age of dam. We present a method that allows including records of calves with unknown age of dam. Assumptions are as follows: (i) foster cows belong to the same breed being evaluated, (ii) there is no correlation between the breeding value (BV) of the calf and the maternal BV of the recipient cow, and (iii) cows of all ages are used as recipients. We examine the issue of bias for the fixed level of unknown age of dam (AOD) and propose an estimator of the effect based on classical measurement error theory (MEM) and a Bayesian approach. Using stochastic simulation under random mating or selection, the MEM estimating equations were compared with BLUP in two situations as follows: (i) full information (FI); (ii) missing AOD information on some dams. Predictions of breeding value (PBV) from the FI situation had the smallest empirical average bias followed by PBV obtained without taking measurement error into account. In turn, MEM displayed the highest bias, although the differences were small. On the other hand, MEM showed the smallest MSEP, for either random mating or selection, followed by FI, whereas ignoring measurement error produced the largest MSEP. As a consequence from the smallest MSEP with a relatively small bias, empirical accuracies of PBV were larger for MEM than those for full information, which in turn showed larger accuracies than the situation ignoring measurement error. It is concluded that MEM equations are a useful alternative for analysing weaning weight data when recipient cows are unknown, as it mitigates the effects of bias in AOD by decreasing MSEP. © 2014 Blackwell Verlag GmbH.

  4. [Anxiety and its inducing factors in men undergoing in vitro fertilization and embryo transfer].

    Science.gov (United States)

    Shu, Ling; Chen, Dan; Jiang, Ling; Zhang, Qiong; Jiang, Li-Ping

    2016-11-01

    To explore the prevalence of anxiety and its inducing factors in men undergoing in vitro fertilization and embryo transfer (IVF-ET). We randomly selected 202 men undergoing IVF-ET in the Infertility and Reproduction Center of the Second Xiangya Hospital of Central South University. On the first day of the IVF-ET cycle, we completed an investigation among the men using a self-designed questionnaire, Self-Rating Anxiety Scale (SAS), Social Support Rating Scale (SSRS), and 3 subscales (marital satisfaction, husband-wife communication, and sexual relationship) of Olson Marital Inventory. Mild anxiety was found in 55 (27.2%) of the included men while the other 147 (72.8%) were non-anxiety males. Compared with the non-anxiety group, the anxiety group showed significant decreases in the total SSRS score (38.65±4.87 vs 36.44±4.21), objective support score (9.22±1.82 vs 8.36±1.18), and utility degree of social support score (6.89±1.50 vs 6.24±1.61) on the first day of the treatment cycle (Psatisfaction (103.04±9.97 vs 96.89±9.90), husband-wife communication (32.29±4.24 vs 30.56±5.43), and sexual relationship (38.03±5.27 vs 34.20±4.41) (Psexual relationship in the men undergoing IVF-ET (P<0.05). The incidence rate of anxiety is high in males undergoing IVF-ET and it is associated with various factors. Psychological aid is needed to these male patients from the staff of the reproduction center.

  5. Proteomic analysis of the Gallus gallus embryo at stage-29 of development.

    Science.gov (United States)

    Agudo, David; Agudo Garcillán, David; Gómez-Esquer, Francisco; Díaz-Gil, Gema; Martínez-Arribas, Fernando; Delcán, José; Schneider, José; Palomar, María Angustias; Linares, Rafael

    2005-12-01

    The chicken (Gallus gallus) is one of the primary models for embryological and developmental studies. In order to begin to understand the molecular mechanisms underlying the normal and abnormal development of the chicken, we used 2-DE to construct a whole-embryo proteome map. Proteins were separated by IEF on IPG strips, and by 11% SDS-PAGE) gels. Protein identification was performed by means of PMF with MALDI-TOF-MS. In all, 105 protein spots were identified, 35 of them implicated in embryo development, 10 related with some diseases, and 16, finally, being proteins that have never been identified, purified or characterized in the chicken before. This map will be updated continuously and will serve as a reference database for investigators, studying changes at the protein level under different physiological conditions.

  6. Silencing CENPF in bovine preimplantation embryo induces arrest at 8-cell stage

    Czech Academy of Sciences Publication Activity Database

    Toralová, Tereza; Šušor, Andrej; Němcová, Lucie; Kepková, Kateřina; Kaňka, Jiří

    2009-01-01

    Roč. 138, č. 5 (2009), s. 783-791 ISSN 1470-1626 R&D Projects: GA ČR GA523/06/1226; GA ČR(CZ) GD204/05/H023 Institutional research plan: CEZ:AV0Z50450515 Keywords : CENPF * bovine embryo * preimplantation development Subject RIV: GI - Animal Husbandry ; Breeding Impact factor: 2.579, year: 2009

  7. Post-fusion treatment with MG132 increases transcription factor expression in somatic cell nuclear transfer embryos in pigs.

    Science.gov (United States)

    You, Jinyoung; Lee, Joohyeong; Kim, Jinyoung; Park, Junhong; Lee, Eunsong

    2010-02-01

    The objective of this study was to examine the effect of post-fusion treatment of somatic cell nuclear transfer (SCNT) oocytes with the proteasomal inhibitor MG132 on maturation promoting factor (MPF) activity, nuclear remodeling, embryonic development, and gene expression of cloned pig embryos. Immediately after electrofusion, SCNT oocytes were treated with MG132 and/or caffeine for 2 hr, vanadate for 0.5 hr, or vanadate for 0.5 hr followed by MG132 for 1.5 hr. Of the MG132 concentrations tested (0-5 microM), the 1 microM concentration showed a higher rate of blastocyst formation (25.9%) than 0 (14.2%), 0.5 (16.9%), and 5 microM (16.9%). Post-fusion treatment with MG132, caffeine, and both MG132 and caffeine improved blastocyst formation (22.1%, 21.4%, and 24.4%, respectively), whereas vanadate treatment inhibited blastocyst formation (6.5%) compared to the control (11.1%). When examined 2 hr after fusion and 1 hr after activation, MPF activity remained at a higher (P fusion with caffeine and/or MG132, but it was decreased by vanadate. The rate of oocytes showing premature chromosome condensation was not altered by MG132 but was decreased by vanadate treatment. In addition, formation of single pronuclei was increased by MG132 compared to control and vanadate treatment. MG132-treated embryos showed increased expression of POU5F1, DPPA2, DPPA3, DPPA5, and NDP52l1 genes compared to control embryos. Our results demonstrate that post-fusion treatment of SCNT oocytes with MG132 prevents MPF degradation and increases expression of transcription factors in SCNT embryos, which are necessary for normal development of SCNT embryos. (c) 2009 Wiley-Liss, Inc.

  8. A catalog of Xenopus tropicalis transcription factors and their regional expression in the early gastrula stage embryo.

    Science.gov (United States)

    Blitz, Ira L; Paraiso, Kitt D; Patrushev, Ilya; Chiu, William T Y; Cho, Ken W Y; Gilchrist, Michael J

    2017-06-15

    Gene regulatory networks (GRNs) involve highly combinatorial interactions between transcription factors and short sequence motifs in cis-regulatory modules of target genes to control cellular phenotypes. The GRNs specifying most cell types are largely unknown and are the subject of wide interest. A catalog of transcription factors is a valuable tool toward obtaining a deeper understanding of the role of these critical effectors in any biological setting. Here we present a comprehensive catalog of the transcription factors for the diploid frog Xenopus tropicalis. We identify 1235 genes encoding DNA-binding transcription factors, comparable to the numbers found in typical mammalian species. In detail, the repertoire of X. tropicalis transcription factor genes is nearly identical to human and mouse, with the exception of zinc finger family members, and a small number of species/lineage-specific gene duplications and losses relative to the mammalian repertoires. We applied this resource to the identification of transcription factors differentially expressed in the early gastrula stage embryo. We find transcription factor enrichment in Spemann's organizer, the ventral mesoderm, ectoderm and endoderm, and report 218 TFs that show regionalized expression patterns at this stage. Many of these have not been previously reported as expressed in the early embryo, suggesting thus far unappreciated roles for many transcription factors in the GRNs regulating early development. We expect our transcription factor catalog will facilitate myriad studies using Xenopus as a model system to understand basic biology and human disease. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Single cell proteomics using frog (Xenopus laevis) blastomeres isolated from early stage embryos, which form a geometric progression in protein content

    OpenAIRE

    Sun, Liangliang; Dubiak, Kyle M.; Peuchen, Elizabeth H.; Zhang, Zhenbin; Zhu, Guijie; Huber, Paul W.; Dovichi, Norman J.

    2016-01-01

    Single cell analysis is required to understand cellular heterogeneity in biological systems. We propose that single cells (blastomeres) isolated from early stage invertebrate, amphibian, or fish embryos are ideal model systems for the development of technologies for single cell analysis. For these embryos, although cell cleavage is not exactly symmetric, the content per blastomere decreases roughly by half with each cell division, creating a geometric progression in cellular content. This pro...

  10. ESTÁDIOS DE DESENVOLVIMENTO DE EMBRIÕES NA OBTENÇÃO DE PLANTAS EM CRUZAMENTOS ENTRE GENITORES APIRENOS DE VIDEIRA EMBRYO GROWTH STAGES ON PLANT OBTENTION FROM CROSSES BETWEEN SEEDLESS GRAPE PARENTS

    Directory of Open Access Journals (Sweden)

    ADRIANE LEITE DO AMARAL

    2001-12-01

    Full Text Available Pesquisou-se o efeito do estádio de desenvolvimento de embriões resgatados na obtenção de plantas em cruzamentos entre genitores apirenos de videira, utilizando dois métodos de cultivo in vitro de sementes-traço. No método 1 (M1, as sementes-traço foram cultivadas 60 dias em meio-de-cultura ER e no método 2 (M2 60 dias em ER, mais 30 dias em meio-de-cultura MS. O estádio de desenvolvimento do embrião foi identificado no final destes períodos, quando do resgate e transferência para meio-de-cultura WP. Quatro tipos de embriões foram identificados: globular, cordiforme, torpedo e indefinido. O estádio globular foi encontrado com maior freqüência, levando à obtenção de maior número de plantas, mesmo tendo apresentado menor capacidade de gerar plantas. A maior eficiência em gerar plantas foi obtida com o estádio torpedo. No M2, ocorreu maior número de embriões germinados e de plantas desenvolvidas. Como o estádio de desenvolvimento do embrião influencia na obtenção de plantas, o resgate de embriões em estádio mais avançado do seu desenvolvimento leva a tem maior capacidade de gerar plantas. Portanto, uso de técnicas que favoreçam o resgate de embriões mais desenvolvidos pode aumentar o número de plantas obtidas.It was investigated the effect of the growth stage of rescued embryos on plant obtention from crosses between seedless grape parents. Two methods for in vitro culture were compared. Method 1 (M1: seed traces were cultured 60 days in ER culture medium. Method 2 (M2: procedure of M1 followed by additional 30 days in MS culture medium. The stage of embryo growth was established at the end of this period, when the embryos were rescued and transfered to WP culture medium. Four classes of embryo stages were indentified: globular, heart, torpedo and undefined. The globular stage, the most frequently found, produced the largest quantity of recovered plants, though it had the lowest capacity of originating plants

  11. Perkembangan Praimplantasi Embrio Mencit dengan Materi Genetik yang Berasal dari Parental, Maternal, dan Inti Sel Somatik (PRE-IMPLANTATION DEVELOPMENT OF MOUSE EMBRYO WITH GENETIC MATERIAL DERIVED FROM PARENTAL, MATERNAL AND SOMATIC CELL NUCLEUS

    Directory of Open Access Journals (Sweden)

    Harry Murti

    2014-05-01

    Full Text Available Cloned embryo and parthenogenetic embryo are a potential source of stem cells for regenerativemedicine. Stem cells derived from those embryos are expected to overcome the ethical issues to the use offertilization embryos for therapeutic purposes. The pre-implantation development is a critical step fordeveloping embryos reach the blastocyst stage. The objectives in vivo of this research are to produce mousecloned embryo, parthenogenetic embryo, and fertilized embryo and to study stages of  in vitro pre-implantation development culture. In vivo fertilized embryos, mouse oocytes, and cumulus cells were usedin this study. Treatment was performed on female mice superovulated with PMSG and hCG injections.Two-cell stage of in vivo fertilized embryos were collected on the second day post hCG injection. Clonedembryos were produced through Somatic Cell Nuclear Transfer (SCNT, which included enucleation, nucleartransfer and artificial activation. Parthenogenetic embryos were produced with artificial activationtechnique. The result of the research indicated that SCNT application was able to produce cloned embryos which could develop to blastocyst stage (3,2%. In addition, artificial activation of oocytes could produceparthenogenetic embryos which were able to develop up to the blastocyst stage (8,6%. In conclusion,efficiency level of parthenogenetic embryos that is able to reach the blastocyst stage was higher than in thecloned embryos. Fertilized embryos shows a better development and more efficient compared to in vitrocloned embryos and parthenogenetic embryos cultures.

  12. Lipofection of siRNA into bovine 8-16-cell stage embryos using zona removal and the well-of-the-well culture system.

    Science.gov (United States)

    Ikeda, Shuntaro; Sugimoto, Miki; Kume, Shinichi

    2018-04-13

    Bovine preimplantation embryos exhibit dramatic biological changes between before and after the 8-16-cell stage. Here we report a simple lipofection method to transfect siRNA into bovine 8-16-cell stage embryos using zona removal and the well-of-the-well (WOW) culture system. Bovine one-cell embryos produced in vitro were freed from the zona pellucida and cultured up to the 8-16-cell stage in WOW dishes. The 8-16-cell embryos were lipofected with siRNA and the transfection efficiency was assessed at 48 h of transfection. Lipofection with a red fluorescent non-targeting siRNA revealed the importance of zona removal for transfection of siRNA into embryos. Using this method, we knocked down the methionine adenosyltransferase 2A (MAT2A) gene, achieving a significant reduction in MAT2A expression (P < 0.05) concomitant with the marked inhibition of blastocyst development. Our proposed method, tentatively named 'Octo-lipofection', may be useful to analyze gene functions in bovine preimplantation embryos without expensive equipment and skill-intensive techniques.

  13. Transcriptome profiles of embryos before and after cleavage in Eriocheir sinensis: identification of developmental genes at the earliest stages

    Science.gov (United States)

    Hui, Min; Cui, Zhaoxia; Liu, Yuan; Song, Chengwen

    2017-07-01

    In crab, embryogenesis is a complicated developmental program marked by a series of critical events. RNA-Sequencing technology offers developmental biologists a way to identify many more developmental genes than ever before. Here, we present a comprehensive analysis of the transcriptomes of Eriocheir sinensis oosperms (Os) and embryos at the 2-4 cell stage (Cs), which are separated by a cleavage event. A total of 18 923 unigenes were identified, and 403 genes matched with gene ontology (GO) terms related to developmental processes. In total, 432 differentially expressed genes (DEGs) were detected between the two stages. Nine DEGs were specifically expressed at only one stage. These DEGs may be relevant to stage-specific molecular events during development. A number of DEGs related to `hedgehog signaling pathway', `Wnt signaling pathway' `germplasm', `nervous system', `sensory perception' and `segment polarity' were identified as being up-regulated at the Cs stage. The results suggest that these embryonic developmental events begin before the early cleavage event in crabs, and that many of the genes expressed in the two transcriptomes might be maternal genes. Our study provides ample information for further research on the molecular mechanisms underlying crab development.

  14. The association between polycystic ovary syndrome and ectopic pregnancy after in vitro fertilization and embryo transfer.

    Science.gov (United States)

    Wang, Jing; Wei, Yongyue; Diao, Feiyang; Cui, Yugui; Mao, Yundong; Wang, Wei; Liu, Jiayin

    2013-08-01

    We sought to assess the association between polycystic ovary syndrome (PCOS) and ectopic pregnancy after in vitro fertilization-embryo transfer (ET). In this retrospective cohort study, we included 5339 women who had clinical pregnancies after in vitro fertilization treatment (PCOS, 205 women; non-PCOS, 5134 women) at Nanjing Medical University (China) between 2007 and 2011. Fresh and cryo-thawed ET cycles were analyzed respectively. The primary outcome measure was the occurrence of ectopic pregnancy. Multivariate logistic regression analysis was used to adjust for important confounders. In fresh ET cycles of women who were undergoing controlled ovarian hyperstimulation (COH; n = 3303), women with PCOS had 3.06 times higher risk of ectopic pregnancy compared with those without PCOS (7.0% vs 2.4%; adjusted odds ratio [aOR], 3.06; 95% confidence interval [CI], 1.34-6.96). In the stratified analysis, for women without PCOS, the high estradiol group (>4085 pg/mL) had higher ectopic pregnancy rates compared with the low estradiol group (≤4085 pg/mL; 3.4% vs 2.0%; aOR, 1.99; 95% CI, 1.19-3.35); however, for women with PCOS, both high and low estradiol groups had high ectopic pregnancy rates (5.6% vs 7.7%; aOR, 0.92; 95% CI, 0.15-5.67). In cryo-thawed ET cycles without COH (n = 2036), the ectopic rates between women with and without PCOS were similar (2.2% vs 2.0%; aOR, 0.94; 95% CI, 0.22-4.07). PCOS was associated with an increased risk of ectopic pregnancy after COH in fresh ET cycles, but not in cryo-thawed ET cycles. A possible explanation is that, compared with women without PCOS, women with PCOS appear to hold a lower threshold of hyperphysiologic estradiol level that triggers the occurrence of ectopic pregnancy after COH. Copyright © 2013 Mosby, Inc. All rights reserved.

  15. Efficacy of dehydroepiandrosterone (DHEA) supplementation for in vitro fertilization and embryo transfer cycles: a systematic review and meta-analysis.

    Science.gov (United States)

    Liu, Yaofang; Hu, Lina; Fan, Lingye; Wang, Fang

    2018-03-01

    Dehydroepiandrosterone (DHEA) supplementation might hold some promise in vitro fertilization and embryo transfer cycles. However, the results remain controversial. We conducted a systematic review and meta-analysis to evaluate the efficacy of DHEA in patients for in vitro fertilization. PubMed, EMbase, Web of science, EBSCO and Cochrane library databases were systematically searched. Randomized controlled trials (RCTs) assessing the effect of DHEA versus placebo on in vitro fertilization were included. Two investigators independently searched articles, extracted data and assessed the quality of included studies. The primary outcomes were clinical pregnancy and live birth rate. Meta-analysis was performed using random-effect model. Six RCTs involving 745 patients were included in the meta-analysis. Overall, compared with placebo, DHEA supplementation was associated with the significant increase in clinical pregnancy (OR = 1.45; 95% CI = 1.04-2.03; p = .03), live birth rate (OR = 2.70; 95% CI = 1.24-5.85; p = .01) and endometrial thickness (Std. mean difference = 0.67; 95% CI = 0.02-1.32; p = .04) but showed no influence on E 2 on hCG day (Std. mean difference = 0.69; 95% CI =  -0.46 to 1.85; p = .24), embryos transferred (Std. mean difference = 0.42; 95% CI =  -0.04 to 0.88; p = .07) and miscarriage rate (OR = 0.43; 95% CI = 0.03-6.66; p = .55). DHEA supplementation could significantly improve clinical pregnancy, live birth rate, endometrial thickness and retrieved oocytes but failed to alter E 2 on hCG day, embryos transferred and miscarriage rate.

  16. Evaluation of an effective multifaceted implementation strategy for elective single-embryo transfer after in vitro fertilization.

    Science.gov (United States)

    Kreuwel, I A M; van Peperstraten, A M; Hulscher, M E J L; Kremer, J A M; Grol, R P T M; Nelen, W L D M; Hermens, R P M G

    2013-02-01

    What is the relationship between the rate of elective single-embryo transfer (eSET) and couples' exposure to different elements of a multifaceted implementation strategy? Additional elements in a multifaceted implementation strategy do not result in an increased eSET rate. A multifaceted eSET implementation strategy with four different elements is effective in increasing the eSET rate by 11%. It is unclear whether every strategy element contributes equally to the strategy's effectiveness. An observational study was performed among 222 subfertile couples included in a previously performed randomized controlled trial. Of the 222 subfertile couples included, 109 couples received the implementation strategy and 113 couples received standard IVF care. A multivariate regression analysis assessed the effectiveness of four different strategy elements on the decision about the number embryos to be transferred. Questionnaires evaluated the experiences of couples with the different elements. Of the couples who received the implementation strategy, almost 50% (52/109) were exposed to all the four elements of the strategy. The remaining 57 couples who received two or three elements of the strategy could be divided into two further classes of exposure. Our analysis demonstrated that additional elements do not result in an increased eSET rate. In addition to the physician's advice, couples rated a decision aid and a counselling session as more important for their decision to transfer one or two embryos, compared with a phone call and a reimbursement offer (P strategy does not always result in an increased effectiveness, which is in concordance with recent literature. This in-depth evaluation of a multifaceted intervention strategy could therefore help to modify strategies, by making them more effective and less expensive.

  17. Necropsy findings in American alligator late-stage embryos and hatchlings from northcentral Florida lakes contaminated with organochlorine pesticides

    Science.gov (United States)

    Sepulveda, M.S.; Del, Piero F.; Wiebe, J.J.; Rauschenberger, H.R.; Gross, T.S.

    2006-01-01

    Increased American alligator (Alligator mississippiensis) embryo and neonatal mortality has been reported from several northcentral Florida lakes contaminated with old-use organochlorine pesticides (OCPs). However, a clear relationship among these contaminants and egg viability has not been established, suggesting the involvement of additional factors in these mortalities. Thus, the main objective of this study was to determine the ultimate cause of mortality of American alligator late-stage embryos and hatchlings through the conduction of detailed pathological examinations, and to evaluate better the role of OCPs in these mortalities. Between 2000 and 2001, 236 dead alligators were necropsied at or near hatching (after ???65 days of artificial incubation and up to 1 mo of age posthatch). Dead animals were collected from 18 clutches ranging in viability from 0% to 95%. Total OCP concentrations in yolk ranged from ???100 to 52,000 ??g/kg, wet weight. The most common gross findings were generalized edema (34%) and organ hyperemia (29%), followed by severe emaciation (14%) and gross deformities (3%). Histopathologic examination revealed lesions in 35% of the animals, with over half of the cases being pneumonia, pulmonary edema, and atelectasis. Within and across clutches, dead embryos and hatchlings compared with their live cohorts were significantly smaller and lighter. Although alterations in growth and development were not related to yolk OCPs, there was an increase in prevalence of histologic lesions in clutches with high OCPs. Overall, these results indicate that general growth retardation and respiratory abnormalities were a major contributing factor in observed mortalities and that contaminants may increase the susceptibility of animals to developing certain pathologic conditions. ?? Wildlife Disease Association 2006.

  18. The overlaying oil type influences in vitro embryo production: differences in composition and compound transfer into incubation medium between oils.

    Science.gov (United States)

    Martinez, Cristina A; Nohalez, Alicia; Parrilla, Inmaculada; Motas, Miguel; Roca, Jordi; Romero, Inmaculada; García-Gonza Lez, Diego L; Cuello, Cristina; Rodriguez-Martinez, Heriberto; Martinez, Emilio A; Gil, Maria A

    2017-09-05

    The oil overlay micro-drop system is widely used for cultures of mammalian gametes and embryos. We evaluated hereby the effects of two unaltered commercial oils- Sigma mineral oil (S-MO) and Nidoil paraffin oil (N-PO)-on in vitro embryo production (IVP) outcomes using a pig model. The results showed that while either oil apparently did not affect oocyte maturation and fertilization rates, S-MO negatively affected embryo cleavage rates, blastocyst formation rates, and, consequently, total blastocyst efficiency of the system. No differences in the oxidation state were found between the oils or culture media incubated under S-MO or N-PO. Although both oils slightly differed in elemental composition, there were no differences in the concentrations of elements between fresh media and media incubated under oils. By contrast, we demonstrated clear oil-type differences in both the composition of volatile organic compounds (VOC) and the transfer of some of these VOC´s (straight-chain alkanes and pentanal and 1,3-diethyl benzene) to the culture medium, which could have influenced embryonic development.

  19. MODELAGEM BIOECONÔMICA DA TRANSFERÊNCIA DE EMBRIÕES EM BOVINOS BIOECONOMIC MODEL IN BOVINE EMBRYO TRANSFER

    Directory of Open Access Journals (Sweden)

    Renato Travassos Beltrame

    2010-04-01

    Full Text Available

    O objetivo deste trabalho foi desenvolver um modelo matemático orientado a eventos de simulação, para auxiliar tomadas de decisão relativas à transferência de embriões em bovinos, considerando-se as dinâmicas de dois componentes da transferência de embriões: receptoras e embriões. Na simulação, não se avaliaram respostas individuais de doadoras a coletas consecutivas e eventos correspondentes na transferência de embriões. Simulou-se o mesmo protocolo para superovulação a todas as doadoras. Receptoras foram sincronizadas simulando-se o uso de prostaglandina. O número de embriões viáveis produzido por doadora e sua variabilidade tiveram como base um processo aleatório de simulação de Monte Carlo, que pressupôs uma distribuição exponencial negativa de densidade de probabilidade. Custos e receitas foram inseridos no modelo por meio de um cenário-base para calcular indicadores econômicos de rentabilidade. A análise sugeriu a impraticabilidade da atividade, se realizada diante do cenário proposto (VPL – R$: 57.596,69. A partir do cenário proposto, o custo médio estimado foi de R$ 1.178,19, e de R$ 980,03, para se obter uma prenhez a partir de uma situação otimizada, sugerida pelo modelo (5/100; 5/190.

    PALAVRAS-CHAVES: Otimização, receptoras, simulação, transferência de embriões, viabilidade econômica.

    A simulation model related to embryo transfer programs in bovine was carried out through a mathematical model directed to events, considering the dynamic of two resources: recipients and embryos. Individual answers of donors to consecutive collections and corresponding events in embryo transfer were not evaluated. The same protocol for superovulation was simulated for all the donor collections, using similar doses of hormones and drugs for all the animals. Recipients were synchronized using prostaglandin. Meantime, the number of viable embryos produced by donor and its variability were based at

  20. Reproductive efficiency of asymptomatic Theileria equi carriers mares submitted to an embryo transfer program

    Directory of Open Access Journals (Sweden)

    Luciana L. Bezerra

    2015-03-01

    Full Text Available This study aimed to assess and evaluate the effects of Theileria equi infection on embryonic recovery, gestation and early embryonic loss. Thirteen Mangalarga Marchador Theileria equi positive donors (diagnosed through nested-PCR and 40 embryos receptors were used. Donors were submitted to two embryo collections in two consecutive estrous cycles (GId; after, the same mares were treated with imidocarb dipropionate (1.2mg/kg IM. in order to collect more embryos in two more estrous cycles (GIId. Receptors were divided into two groups (control and with treated with 20 animals each, where one group was the control (GIr and the other one (GIIr treated with 1.2mg/kg IM of imidocarb dipropionate assessing the gestation rate at 15, 30, 45 and 60 days. After 52 embryo collections, the embryonic recovery rates were 53.84% (14/26 and 65.38% (17/26 (p> 0.05 for GId and GIId, respectively. The gestation rate was 70% (14/20 (p>0.05 at 15, 30, 45 and 60 days in group GIr and for GIIr was 85% (17/20 (p>0.05 at 15 days, 80% (16/20 (p>0.05 at 30, 45 and 60 days. The treatment with imidocarb dipropionate did not cause significant improvement in the reproductive efficiency at an ET program.

  1. Predictive value of plasma human chorionic gonadotropin measured 14 days after Day-2 single embryo transfer

    DEFF Research Database (Denmark)

    Løssl, Kristine; Oldenburg, Anna; Toftager, Mette

    2017-01-01

    -2 embryos. The aim of the present study was to investigate the predictive value of p-hCG measured exactly 14 days after the most commonly used Day-2 SET on pregnancy, delivery, and perinatal outcome. Material and methods: A retrospective analysis of prospectively collected data on 466 women who had...

  2. Application of in vitro production-embryo transfer in the protection ...

    African Journals Online (AJOL)

    Yomi

    2012-01-12

    Jan 12, 2012 ... This study was carried out to investigate if the technology of in vitro embryo production can be used for protection and development of milk productivity potential of the superior cattle via ablation. The ovaries of the superior cattle selected for ablation were collected immediately after slaughter. Bovine.

  3. Monochorionic-triamniotic triplet pregnancy after intracytoplasmic sperm injection, assisted hatching, and two-embryo transfer: first reported case following IVF

    Directory of Open Access Journals (Sweden)

    Eller Daniel P

    2003-08-01

    Full Text Available Abstract Background We present a case of monochorionic-triamniotic pregnancy that developed after embryo transfer following in vitro fertilization (IVF. Methods After controlled ovarian hyperstimulation and transvaginal retrieval of 22 metaphase II oocytes, fertilization was accomplished with intracytoplasmic sperm injection (ICSI. Assisted embryo hatching was performed, and two embryos were transferred in utero. One non-transferred blastocyst was cryopreserved. Results Fourteen days post-transfer, serum hCG level was 423 mIU/ml and subsequent transvaginal ultrasound revealed a single intrauterine gestational sac with three separate amnion compartments. Three distinct foci of cardiac motion were detected and the diagnosis was revised to monochorionic-triamniotic triplet pregnancy. Antenatal management included cerclage placement at 19 weeks gestation and hospital admission at 28 weeks gestation due to mild preeclampsia. Three viable female infants were delivered via cesarean at 30 5/7 weeks gestation. Conclusions The incidence of triplet delivery in humans is approximately 1:6400, and such pregnancies are classified as high-risk for reasons described in this report. We also outline an obstetric management strategy designed to optimize outcomes. The roles of IVF, ICSI, assisted embryo hatching and associated laboratory culture conditions on the subsequent development of monozygotic/monochorionic pregnancy remain controversial. As demonstrated here, even when two-embryo transfer is employed after IVF the statistical probability of monozygotic multiple gestation cannot be reduced to zero. We encourage discussion of this possibility during informed consent for the advanced reproductive technologies.

  4. IVF policy and global/local politics: the making of multiple-embryo transfer regulation in Taiwan.

    Science.gov (United States)

    Wu, Chia-Ling

    2012-08-01

    This paper analyzes the regulatory trajectory of multiple-embryo transfer in in-vitro fertilization (IVF) in Taiwan. Taking a latecomer to policy-making as the case, it argues the importance of conceptualizing the global/local dynamics in policy-making for assisted reproductive technology (ART). The conceptual framework is built upon recent literature on standardization, science policy, and global assemblage. I propose three interrelated features that reveal the "global in the local": (1) the power relationships among stakeholders, (2) the selected global form that involved actors drew upon, and (3) the re-contextualized assemblage made of local networks. Data included archives, interviews, and participant observation. In different historical periods the specific stakeholders selected different preferred global forms for Taiwan, such as Britain's code of ethics in the 1990s, the American guideline in the early 2000s, and the European trend in the mid-2000s. The global is heterogeneous. The failure to transfer the British regulation, the revision of the American guideline by adding one more embryo than it specified, and the gap between the cited European trend and the "no more than four" in Taiwan's 2007 Human Reproduction Law all show that the local network further transforms the selected global form, confining it to rhetoric only or tailoring it to local needs. Overall, Taiwanese practitioners successfully maintained their medical autonomy to build a 'flexible standardization'. Multiple pregnancy remains the most common health risk of IVF in Taiwan. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Embryo transfer in no cycling Crioula and Quarter horse breeds treated with estradiol cipionate and long-acting progesterone

    Directory of Open Access Journals (Sweden)

    Fernanda Kaercher

    2011-12-01

    Full Text Available The aim of this work was to prepare the mares for embryo transfer. In group 1 (G1,treated, n=15, recipient mares in anoestrus or in a transition period were treated with 5.0, 3.0 and 2.0 mg of estradiol cipionate at the days 0, 1 and 2 respectively, beginning at the day of ovulation (DO. From the fourth day on, the mares this group received long-acting progesterone weekly, up to the 120 day of gestation. At D8, the embryo was collected from the donor and transferred to the recipient. At D12, the ultrasonographyc diagnosis of pregnancy was carried out. The control group (G2, not treated, n=20 was formed by cycling recipient mares, displaying ovulation at each 2 to 3 days after the donors mare ovulation. The pregnancy rate was higher (p<0.05 in the mares from G2 (85.0% than from G1 (53.3%. Thus, it could be concluded that the treated mares although showed lesser pregnancy rate than the cycling mare, were satisfactory alternative to be used mainly when there is no available cycling recipient.

  6. The heat transfer analysis of the first stage blade

    International Nuclear Information System (INIS)

    Hong, Yong Ju; Choi, Bum Seog; Park, Byung Gyu; Yoon, Eui Soo

    2001-01-01

    To get higher efficiency of gas turbine, the designer should have more higher Turbine Inlet Temperature(TIT). Today, modern gas turbine having sophisticated cooling scheme has TIT above 1,700 .deg. C. In the Korea, many gas turbine having TIT above 1,300 .deg. C was imported and being operated, but the gas with high TIT above 1,300 .deg. C in the turbine will give damage to liner of combustor, and blade of turbine and etc. So frequently maintenance for parts enduring high temperature was performed. In this study, the heat transfer analysis of cooling air in the internal cooling channel (network analysis) and temperature analysis of the blade (Finite Element Analysis) in the first stage rotor was conducted for development of the optimal cooling passage design procedure. The results of network analysis and FEM analysis of blade show that the high temperature spot are occurred at the leading edge, trailing edge near tip, and platform. So to get more reliable performance of gas turbine, the more efficient cooling method should be applied at the leading edge and tip section and the thermal barrier coating on the blade surface has important role in cooling blade

  7. Transfer of egg white proteins and activation of proteases during the development of Anas platyrhynchos domestica embryo.

    Science.gov (United States)

    Shbailat, Seba Jamal; Abuassaf, Razan Ataallah

    2018-03-01

    The route of egg white transfer into the yolk and the mechanisms underlying the digestion of egg proteins are unexplored in the fertilized egg of the duck, Anas platyrhynchos domestica. Here, we investigated the route(s) of egg white transfer and we determined the type of activated proteases during duck embryo development. Initially, we tested the electrophoretic patterns of egg proteins throughout development. Then, we used lysozyme as a reference protein to follow egg white transfer and we measured its activity. After that, we determined the type of activated proteases by employing different types of protease inhibitors. Several presumptive egg white protein bands appeared in different egg compartments. Also, lysozyme activity was detected chronologically on day 15 in the extraembryonic fluid, on day 17 in the amniotic and intestinal fluids and on day 19 in the yolk. Furthermore, acidic aspartic proteases seemed to be activated at hatch in the intestine and late in development in the yolk. Our results suggest that the main route of egg white transfer into the yolk is through the amniotic cavity and intestinal lumen. Also, the transferred egg white and endogenous yolk proteins are probably digested by the activated acidic proteases in the intestine and yolk.

  8. Effects of the ionizing radiations in the different stages of the embryo-fetal development

    International Nuclear Information System (INIS)

    Perez, Maria R.

    2001-01-01

    'In utero' exposures to ionizing radiation are a very important subject in radiological protection concerning not only the prevention but also the estimation of the associated risks. In these situations the perception of risks by the pregnant woman and the involved professionals could not always be correlated with their objective magnitude. In this communication we describe the effects of prenatal exposure to ionizing radiation, the thresholds and their relation with the gestational age, taking into account occupationally exposed women, patients undergoing medical procedures and public members. The dose estimation, the evaluation of the potential associated risks and the relation with the spontaneous incidence of the considered effects are analyzed in the framework of the basic principles of radiological protection. Most of diagnostic procedures properly done do not imply induction of deterministic effects in embryo/fetus. Therapeutical procedures and accidental overexposures could be associated with significant risks of deterministic effects. Childhood cancer induction is an stochastic effect without threshold and every 'in utero' exposure will increase their probability. (author)

  9. Runx expression is mitogenic and mutually linked to Wnt activity in blastula-stage sea urchin embryos.

    Directory of Open Access Journals (Sweden)

    Anthony J Robertson

    Full Text Available The Runt homology domain (Runx defines a metazoan family of sequence-specific transcriptional regulatory proteins that are critical for animal development and causally associated with a variety of mammalian cancers. The sea urchin Runx gene SpRunt-1 is expressed throughout the blastula stage embryo, and is required globally during embryogenesis for cell survival and differentiation.Depletion of SpRunt-1 by morpholino antisense-mediated knockdown causes a blastula stage deficit in cell proliferation, as shown by bromodeoxyuridine (BrdU incorporation and direct cell counts. Reverse transcription coupled polymerase chain reaction (RT-PCR studies show that the cell proliferation deficit is presaged by a deficit in the expression of several zygotic wnt genes, including wnt8, a key regulator of endomesoderm development. In addition, SpRunt-1-depleted blastulae underexpress cyclinD, an effector of mitogenic Wnt signaling. Blastula stage cell proliferation is also impeded by knockdown of either wnt8 or cyclinD. Chromatin immunoprecipitation (ChIP indicates that Runx target sites within 5' sequences flanking cyclinD, wnt6 and wnt8 are directly bound by SpRunt-1 protein at late blastula stage. Furthermore, experiments using a green fluorescent protein (GFP reporter transgene show that the blastula-stage operation of a cis-regulatory module previously shown to be required for wnt8 expression (Minokawa et al., Dev. Biol. 288: 545-558, 2005 is dependent on its direct sequence-specific interaction with SpRunt-1. Finally, inhibitor studies and immunoblot analysis show that SpRunt-1 protein levels are negatively regulated by glycogen synthase kinase (GSK-3.These results suggest that Runx expression and Wnt signaling are mutually linked in a feedback circuit that controls cell proliferation during development.

  10. In vitro development competence of bovine nuclear transfer embryos derived from Nanog-overexpressing fibroblast cells

    Directory of Open Access Journals (Sweden)

    Xi-bang Zheng, Yan Yun, Yong-ce Hu, Yong Li, Hua-yan Wang, Xiao-ling Ma, Jin-qiang Sui, An-min Lei and Zhong-ying Dou

    2014-04-01

    Full Text Available The purpose of this study was to establish Nanog-expressing cell lines that can be used as donor cells to construct transgenic cloned embryos, and to investigate their in vitro development competence. By reverse transcription-polymerase chain reaction (RT-PCR, the cDNA of Nanog gene was cloned from fetal bovine primordial genital ridge tissues. The gene was inserted into PMD18-T vector using recombination techniques and then subcloned into vector pEGFP-C1. After confirmation by restrictive endonuclease digestion and sequencing, the recombinant plasmid pEGFP-Nanog was transfected into skin fibroblast cells. A stable transfected cell line was successfully established after two months of selection with neomycine (G418. Fluorescence microscopy, RT-PCR, and Western Blotting assays indicated that Nanog mRNA and EGFP-Nanog fusion protein were expressed in these cells. The EGFP-Nanog expressing fibroblast cells and the intact fibroblast cells (BEF422 were respectively used to construct cloned embryos. The results showed that the cleavage rate of recombinant embryos in BEF422 cells was significantly (P<0.05 higher than in EGFP-Nanog expressing cells (82.14 vs 40.38 %, but the blastocyst development rate in the latter was slightly higher than in the former (17.30 vs 14.29% (P<0.05, indicating that Nanog-overexpressed fibroblasts may be a better candidate of donor cells. To our knowledge, this is the first time that Nanog gene has been introduced into fibroblast cells to produce cloned embryos in bovine.

  11. Confinement and clearance of OCT4 in the porcine embryo at stereomicroscopically defined stages around gastrulation

    DEFF Research Database (Denmark)

    Vejlsted, Morten; Offenberg, Hanne Kjær; Thorup, Flemming

    2006-01-01

    maintaining expression of OCT4 at the end of gastrulation. In the ectodermal and mesodermal cell lineages, OCT4 became undetectable at the neural groove and somite stage, respectively. As in the mouse, PGCs showed onset of c-kit expression when located in extraembryonal compartments. They appeared to follow...... the endoderm during extraembryonal allocation and the mesoderm on return to the genital ridge....

  12. Developmental toxicity and induction of vitellogenin in embryo-larval stages of zebrafish (Danio rerio) exposed to methyl Paraben.

    Science.gov (United States)

    Dambal, Vrinda Yatin; Selvan, Karthikeyan Ponnien; Lite, Christy; Barathi, S; Santosh, Winkins

    2017-07-01

    MethylParaben (MP), a methyl ester of p-hydroxybenzoic acid, is used as an anti-microbial preservative in foods, drugs and cosmetics for decades. It enters the aquatic environment, and can have toxic effects on aquatic organisms. Little is known on the developmental toxicity of MP exposure to zebrafish during early life stages. In this study, the developmental effects of MP were evaluated in embryo-larval zebrafish (at concentrations ranging from 100μM, 200μM, 400µM, 800μM and 1000μM for 96h post fertilization (hpf). The survival, hatching, heart beat rate and developmental abnormalities were observed in the embryos exposed to MP. MP exposure resulted in decreased heart rate and hatching rate. Defects including pericardial edema blood cell accumulation and bent spine were observed in all the treated concentration, except at 100μM. With increasing concentrations, the frequency of these defects increased. The 96 hpf LC 50 of MP was calculated to be 428μM (0.065mg/L). Furthermore, RT-PCR result showed that in larval zebrafish exposed to 100μM (0.015mg/L) of MP till 96 hpf, expression of vitellogenin I (Vtg -I) was significantly upregulated compared to the control group. This data suggest that even though lower concentrations of MP do not cause phenotypic malformations, it leads to dysregulated expression of estrogenic biomarker gene Vtg-I. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Transferência de blastocisto após descongelamento de embriões em mórula resultando em gestação gemelar: relato de caso Twin pregnancy after thawing of morula embryos and blastocyst transfer: case report

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo T. Albuquerque

    1999-12-01

    Full Text Available A criopreservação de embriões em estágios mais tardios do desenvolvimento parece apresentar resultados satisfatórios. Com o objetivo de melhor testar a sobrevivência e o desenvolvimento de embriões, os mesmos foram criopreservados e descongelados em estadio de mórula ou blastocisto e deixados em cultura para que pudesse ser avaliada sua evolução natural. Dos 2 blastocistos e 5 mórulas congelados, 4 mórulas sobreviveram ao descongelamento, tendo sido transferidas em estadio de blastocisto, 24 horas depois. A transferência, realizada em paciente jovem, segundo casamento de homem vasectomizado há dez anos, resultou em gestação gemelar. O descongelamento de embriões em estadio de mórula e a observação in vitro da retomada de seu desenvolvimento até o estadio de blastocisto fornecem um parâmetro adicional na avaliação da qualidade do embrião e, provavelmente, melhore as taxas de gravidez.The cryopreservation of embryos in late developing stages seems to present satisfactory results. With the purpose of better testing the embryos' survival, they were cryopreserved in the morula or blastocyst stage, thawed and left in culture for 24 hours so that their natural evolution could be observed. Amongst the frozen 2 blastocysts and 5 morulas, 4 morulas survived the thawing process, being transferred as blastocysts 24 hours later. The transfer was performed in a young patient, second marriage of a ten-year vasectomized man and resulted in twin pregnancy. Thawing morula embryos and the in vitro observation of their development resumption until the blastocyst stage give us an additional parameter in the quality evaluation of the embryo and probably an improvement in pregnancy rates.

  14. Influence of temperature and humidity in a equine embryo transfer program, in the Baixada Fluminense, Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    Jhonnatha Paulo Oliveira

    2015-06-01

    Full Text Available ABSTRACT. Oliveira J.P., Jacob J.C.F., Jesus V.L.T. & Silva P.C.A. [Influence of temperature and humidity in a equine embryo transfer program, in the Baixada Fluminense, Rio de Janeiro.] Influência da temperatura e umidade ambiente em um programa de transferência de embriões equinos, na Baixada Fluminense, Rio de Janeiro. Revista Brasileira de Medicina Veterinária, 37(2:158- 162, 2015. Departamento de Reprodução e Avaliação Animal, Universidade Federal Rural do Rio de Janeiro, BR 465, Km 7, Seropédica, RJ 23890-970, Brasil. E-mail: juliorep@ufrrj.br This study aimed to evaluate the relationship between high environment temperature and humidity and reproductive rates in a equine embryo transfer program in Baixada Fluminense RJ. We evaluated the reproductive history of 60 donor mares and 111 recipient mares during summer of breeding seasons of 2008/2009, 2009/2010 and 2010/2011. Daily climatics data of environmental temperature (°C and relative humidity (% for the each breeding seasons were obtained from the web site of the National Institute of Meteorology (INMET, based on these data we calculated the temperature x humidity index (TUI wich measures the thermal comfort zone. The reproductive parameters assessed were embryo recovery rate (RR and pregnancy rate (PR. After the computation of reproductive and climatic data, these were compared to establish relationships between high temperatures and humidity on reproductive rates. There was a negative relationship between RR and high environmental temperatures, especially in the summer time, greater RR at 26°C (71% and lower RR at 27°C (51.4% (p <0.05. To PR there was a negative relationship to high environmental temperatures, the higher PR was obtained at 24°C (81.5% and lowest PG (35% at 27°C (p <0.05. We conclude that there are relationships between environmental variables and ET success.

  15. Annotating and quantifying pri-miRNA transcripts using RNA-Seq data of wild type and serrate-1 globular stage embryos of Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Daniel Lepe-Soltero

    2017-12-01

    Full Text Available The genome annotation for the model plant Arabidopsis thaliana does not include the primary transcripts from which MIRNAs are processed. Here we present and analyze the raw mRNA sequencing data from wild type and serrate-1 globular stage embryos of A. thaliana, ecotype Columbia. Because SERRATE is required for pri-miRNA processing, these precursors accumulate in serrate-1 mutants, facilitating their detection using standard RNA-Seq protocols. We first use the mapping of the RNA-Seq reads to the reference genome to annotate the potential primary transcripts of MIRNAs expressed in the embryo. We then quantify these pri-miRNAs in wild type and serrate-1 mutants. Finally, we use differential expression analysis to determine which are up-regulated in serrate-1 compared to wild type, to select the best candidates for bona fide pri-miRNAs expressed in the globular stage embryos. In addition, we analyze a previously published RNA-Seq dataset of wild type and dicer-like 1 mutant embryos at the globular stage [1]. Our data are interpreted and discussed in a separate article [2].

  16. Annotating and quantifying pri-miRNA transcripts using RNA-Seq data of wild type and serrate-1 globular stage embryos of Arabidopsis thaliana.

    Science.gov (United States)

    Lepe-Soltero, Daniel; Armenta-Medina, Alma; Xiang, Daoquan; Datla, Raju; Gillmor, C Stewart; Abreu-Goodger, Cei

    2017-12-01

    The genome annotation for the model plant Arabidopsis thaliana does not include the primary transcripts from which MIRNAs are processed. Here we present and analyze the raw mRNA sequencing data from wild type and serrate-1 globular stage embryos of A. thaliana , ecotype Columbia. Because SERRATE is required for pri-miRNA processing, these precursors accumulate in serrate-1 mutants, facilitating their detection using standard RNA-Seq protocols. We first use the mapping of the RNA-Seq reads to the reference genome to annotate the potential primary transcripts of MIRNAs expressed in the embryo. We then quantify these pri-miRNAs in wild type and serrate-1 mutants. Finally, we use differential expression analysis to determine which are up-regulated in serrate-1 compared to wild type, to select the best candidates for bona fide pri-miRNAs expressed in the globular stage embryos. In addition, we analyze a previously published RNA-Seq dataset of wild type and dicer-like 1 mutant embryos at the globular stage [1]. Our data are interpreted and discussed in a separate article [2].

  17. Embryonic death, dwarfism and fetal malformations after irradiation of embryos at the zygote stage. Studies on two mouse strains

    International Nuclear Information System (INIS)

    Jacquet, P.; Saint-Georges, L. de; Baugnet-Mahieu, L.; Vankerkom, J.

    1995-01-01

    Female mice of the BALB/c and CF1 strains were mated and irradiated with various doses of X-rays 7 h after presumed fertilization. 18 days later, females were killed and their uteri examined for prenatal mortality at the different stages of development. Living fetuses were weighed and examined for the presence of external malformations. A number of them were also examined for skeletal anomalies. Radiation induced mainly a dose-dependent increase of the preimplantation loss in the BALB/c strain and of the early postimplantation loss in the CF1 strain. Embryos of the BALB/c strain were refractory to the induction of teratogenic effects after such preimplantation irradiation. In CF1 mice, the frequency of malformed fetuses increased regularly after irradiation, the difference with controls being significant for the doses of 10, 50 and 100 cGy. Dwarfism occurrence also appeared to be increased by irradiation in this strain, although the importance of this effect varied depending on the criterion chosen for the assessment of dwarfs. With the definition proposed in the present paper, the increase in the frequency of dwarfs paralleled that of malformed fetuses, being significant after doses of 50 and 100 cGy. Irradiation did not increase the frequency of skeletal anomalies. A careful examination of the various data obtained to date led us to conclude that radiation may possibly be teratogenic in several mouse strains, when administered as early as during the one-cell stage and, to a lesser extent, during the following preimplantation stages. However, early prenatal mortality will remain by far the greatest risk associated with an exposure to radiation during this period. Moreover, the relativity of the risk of abnormality due to such irradiation should be considered in the context of the high prevalence of developmental defects spontaneously occurring during human pregnancy

  18. Embryonic death, dwarfism and fetal malformations after irradiation of embryos at the zygote stage. Studies on two mouse strains

    Energy Technology Data Exchange (ETDEWEB)

    Jacquet, P.; Saint-Georges, L. de; Baugnet-Mahieu, L. [Laboratory of Radiobiology, Department of Radioprotection, CEN/SCK, Mol (Belgium); Vankerkom, J. [Division of Environmental Research, VITO, Mol (Belgium)

    1995-11-01

    Female mice of the BALB/c and CF1 strains were mated and irradiated with various doses of X-rays 7 h after presumed fertilization. 18 days later, females were killed and their uteri examined for prenatal mortality at the different stages of development. Living fetuses were weighed and examined for the presence of external malformations. A number of them were also examined for skeletal anomalies. Radiation induced mainly a dose-dependent increase of the preimplantation loss in the BALB/c strain and of the early postimplantation loss in the CF1 strain. Embryos of the BALB/c strain were refractory to the induction of teratogenic effects after such preimplantation irradiation. In CF1 mice, the frequency of malformed fetuses increased regularly after irradiation, the difference with controls being significant for the doses of 10, 50 and 100 cGy. Dwarfism occurrence also appeared to be increased by irradiation in this strain, although the importance of this effect varied depending on the criterion chosen for the assessment of dwarfs. With the definition proposed in the present paper, the increase in the frequency of dwarfs paralleled that of malformed fetuses, being significant after doses of 50 and 100 cGy. Irradiation did not increase the frequency of skeletal anomalies. A careful examination of the various data obtained to date led us to conclude that radiation may possibly be teratogenic in several mouse strains, when administered as early as during the one-cell stage and, to a lesser extent, during the following preimplantation stages. However, early prenatal mortality will remain by far the greatest risk associated with an exposure to radiation during this period. Moreover, the relativity of the risk of abnormality due to such irradiation should be considered in the context of the high prevalence of developmental defects spontaneously occurring during human pregnancy.

  19. A simplified table for staging embryos of the pipid frog Pipa arrabali

    Directory of Open Access Journals (Sweden)

    OLÍVIA G.S. ARAÚJO

    Full Text Available ABSTRACT Pipa is a Neotropical genus of frogs that dwell in freshwater environments. It includes four species that lack free-swimming larvae (P. aspera, P. arrabali, P. pipa, and P. snethlageae and three with tadpoles (P. carvalhoi, P. myersi, and P. parva. Developmental tables such as the one proposed by Nieuwkoop and Faber might be useful for Pipa species with tadpoles. However, for the other Pipa species, to determine stages by this table or by any of the tables already prepared for frogs without tadpoles (e.g., Crinia nimbus, Eleutherodactylus coqui, and Oreobates barituensis is impossible. By using embryonic, juvenile, and subadult specimens, we generated a staging table for P. arrabali, from the moment limb buds were first observed until birth, based on diagnostic features such as snout-vent length; growth, morphology, and reabsorption of the external tail; growth and differentiation of fore and hind limbs; development of intestine and vent tube; position of the angle of the mouth relative to nostrils and eyes; and color of preserved individuals. Based on these observations, we discuss some noteworthy traits (e.g., posture of hands and feet. We also compare the pattern of development of P. arrabali with that of other anuran species (with and without tadpoles.

  20. Risk assessment of porcine reproductive and respiratory syndrome virus (PRRSV) transmission via somatic cell nuclear transfer (SCNT) embryo production using oocytes from commercial abattoirs.

    Science.gov (United States)

    Gregg, K; Xiang, T; Arenivas, S S; Hwang, E; Arenivas, F; Chen, S-H; Walker, S; Picou, A; Polejaeva, I

    2011-05-01

    Somatic cell nuclear transfer (SCNT) technology has become a powerful tool for reproductive biology to preserve and propagate valuable genetics for livestock. Embryo production through SCNT involves enucleation of the oocyte and insertion of a somatic donor cell into the oocyte. These procedures lead to a few small openings on the zona pellucida that may elevate risk of viral infection for the produced SCNT embryos. The oocytes used for SCNT are mainly obtained from abattoirs where viral contamination is almost inevitable. Therefore, a systematic evaluation of risk of disease transmission through SCNT embryo production is necessary prior large scale implementation of this technology in the livestock industry. The objective of the current study was to evaluate the risk of disease transmission via SCNT embryo production and transfer by testing for the presence of porcine reproductive and respiratory syndrome virus (PRRSV) throughout the process of SCNT embryo production. The presence of PRRSV in each step of SCNT embryo production, from donor cells to pre-implantation SCNT embryo culture, was carefully examined using a real-time PCR assay with a sensitivity of five copies per-reaction. All 114 donor cell lines derived from pig skin tissue over a period of 7 years in our facility tested negative for PRRSV. Out of the 68 pooled follicular fluid samples collected from 736 ovaries, only four (5.9%) were positive indicating a small amount of viral molecule present in the oocyte donor population. All 801 Day 7 SCNT embryos produced in four separate trials and over 11,571 washed oocytes obtained in 67 batches over 10 months tested negative. These oocytes were collected from multiple abattoirs processing animals from areas with high density of pig population and correspond to a donor population of over 5828 individuals. These results indicate that the oocytes from abattoirs were free of PRRSV infection and therefore could be safely used for in vitro embryo production

  1. Cryo-thawed embryo transfer: natural versus artificial cycle. A non-inferiority trial.(ANTARCTICA trial

    Directory of Open Access Journals (Sweden)

    Groenewoud Eva R

    2012-09-01

    Full Text Available Abstract Background Frozen thawed embryo transfer (FET is a cost- effective adjunct to IVF or IVF-ICSI treatment. In order to optimize treatment outcome, FET should be carried out during a period of optimal endometrial receptivity. To optimize implantation several methods for endometrium preparation have been proposed. In natural cycle FET (NC-FET, the endometrium develops under endogenous hormonal stimulation. The development of the dominant follicle and endometrium is monitored by ultrasound and FET is timed after triggering ovulation induction or determination of the spontaneous LH surge. In an artificial cycle FET (AC-FET estrogens and progesterone are administered to prepare the endometrium for implantation. While the currently available data show no significant difference in pregnancy rates between these methods, well designed randomized controlled trials are lacking. Moreover there is little literature on difference in cancellation rates, cost-efficiency and adverse events. Methods and design In this randomized, multi-centre, non-inferiority trial we aim to test the hypothesis that there is no significant difference in live birth rates between patients undergoing NC-FET versus AC-FET. The primary outcome will be live birth rate per embryo transfer procedure. Secondary outcomes will be ongoing and clinical pregnancy rate, cancellation rate, (serious adverse events and cost-efficiency. Based on a live birth rate of 20% and a minimal clinical important difference of 7,5% (one-sided alpha 2,5%, beta 20% a total of 1150 patients will be needed. Analyzes will be performed using both per protocol as well as intention to treat analyses. Discussion This prospective, randomized, non –inferiority trial aims to address the hypothesis that there is no significant difference in live birth rates between patients undergoing NC-FET versus patients undergoing AC-FET. Moreover it addresses cost-efficiency as well as the perceived burden of both treatments

  2. Embryo transfer techniques: an American Society for Reproductive Medicine survey of current Society for Assisted Reproductive Technology practices.

    Science.gov (United States)

    Toth, Thomas L; Lee, Malinda S; Bendikson, Kristin A; Reindollar, Richard H

    2017-04-01

    To better understand practice patterns and opportunities for standardization of ET. Cross-sectional survey. Not applicable. Not applicable. An anonymous 82-question survey was emailed to the medical directors of 286 Society for Assisted Reproductive Technology member IVF practices. A follow-up survey composed of three questions specific to ET technique was emailed to the same medical directors. Descriptive statistics of the results were compiled. The survey assessed policies, protocols, restrictions, and specifics pertinent to the technique of ET. There were 117 (41%) responses; 32% practice in academic settings and 68% in private practice. Responders were experienced clinicians, half of whom had performed technique. Multiple steps in the ET process were identified as "highly conserved;" others demonstrated discordance. ET technique is divided among [1] trial transfer followed immediately with ET (40%); [2] afterload transfer (30%); and [3] direct transfer without prior trial or afterload (27%). Embryos are discharged in the upper (66%) and middle thirds (29%) of the endometrial cavity and not closer than 1-1.5 cm from fundus (87%). Details of each step were reported and allowed the development of a "common" practice ET procedure. ET training and practices vary widely. Improved training and standardization based on outcomes data and best practices are warranted. A common practice procedure is suggested for validation by a systematic literature review. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  3. 45,X product of conception after preimplantation genetic diagnosis and euploid embryo transfer: evidence of a spontaneous conception confirmed by DNA fingerprinting.

    Science.gov (United States)

    Bettio, Daniela; Capalbo, Antonio; Albani, Elena; Rienzi, Laura; Achille, Valentina; Venci, Anna; Ubaldi, Filippo Maria; Levi Setti, Paolo Emanuele

    2016-09-06

    Preimplantation genetic screening (PGS) provides an opportunity to eliminate a potential implantation failure due to aneuploidy in infertile couples. Some studies clearly show that twins following single embryo transfer (SET) can be the result of a concurrent natural conception and an incidence as high as 1 in 5 twins has been reported. In our case PGS was performed on trophectoderm (TE) biopsies by quantitative polymerase chain reaction (qPCR). The product of conception (POC) was cytogenetically investigated after selection of the placental villi by means of the direct method. Molecular cytogenetic characterization of the POC was performed by fluorescence in situ hybridization (FISH) and array-comparative genomic hybridization (a-CGH) analyses. To investigate the possibility of a spontaneous conception, a panel of 40 single nucleotide polymorphisms (SNPs) was used to compare genetic similarity between the DNA of the POC and the DNA leftover of the TE biopsy. We describe a 36-year old infertile woman undergoing PGS who had a spontaneous abortion after a single euploid embryo transfer on a spontaneous cycle. The POC showed a 45,X karyotype confirmed by FISH and a-CGH. DNA fingerprinting demonstrated a genetic similarity of 75 % between the DNA of the POC and TE biopsy, consistent with a sibling status. All supernumerary euploid embryos were also tested showing a non-self relationship with the POC, excluding a mix-up event at the time of fetal embryo transfer. DNA fingerprinting of the transferred blastocyst and POC, confirmed the occurrence of a spontaneous conception. This case challenges the assumption that a pregnancy after assisted reproductive technology (ART) is always a result of ART, and strengthens the importance to avoid intercourses during PGS and natural transfer cycles. Moreover, cytogenetic analysis of the POCs is strongly recommended along with fingerprinting children born after PGS to see what the concordance is between the embryo transferred and

  4. Genomic selection strategies in dairy cattle breeding programmes: Sexed semen cannot replace multiple ovulation and embryo transfer as superior reproductive technology

    DEFF Research Database (Denmark)

    Pedersen, Louise Dybdahl; Kargo, Morten; Berg, Peer

    2012-01-01

    The aim of this study was to test whether the use of X-semen in a dairy cattle population using genomic selection (GS) and multiple ovulation and embryo transfer (MOET) increases the selection intensity on cow dams and thereby the genetic gain in the entire population. Also, the dynamics of using...

  5. Evaluation of Closed Adult Nucleus Multiple Ovulation and Embryo Transfer and Conventional Progeny Testing Breeding Schemes for Milk Production from Crossbred Cattle in the Tropics

    NARCIS (Netherlands)

    Kosgey, I.S.; Kahi, A.K.; Arendonk, van J.A.M.

    2005-01-01

    The potential benefits of closed adult nucleus multiple ovulation and embryo transfer (MOET) and conventional progeny testing (CNS) schemes, and the logistics of their integration into large-scale continuous production of crossbred cattle were studied by deterministic simulation. The latter was

  6. Detection of endometrial and subendometrial vasculature on the day of embryo transfer and prediction of pregnancy during fresh in vitro fertilization cycles

    Directory of Open Access Journals (Sweden)

    Ari Kim

    2014-09-01

    Conclusion: Three dimensional PD-US was a useful and effective method for assessing endometrial blood flow in IVF cycles. Good endometrial blood flow on the day of embryo transfer might be associated with high pregnancy success with a GnRH long protocol, because this is indicative of endometrial receptivity in fresh IVF cycles.

  7. Analysis of compaction initiation in human embryos by using time-lapse cinematography.

    Science.gov (United States)

    Iwata, Kyoko; Yumoto, Keitaro; Sugishima, Minako; Mizoguchi, Chizuru; Kai, Yoshiteru; Iba, Yumiko; Mio, Yasuyuki

    2014-04-01

    To analyze the initiation of compaction in human embryos in vitro by using time-lapse cinematography (TLC), with the goal of determining the precise timing of compaction and clarifying the morphological changes underlying the compaction process. One hundred and fifteen embryos donated by couples with no further need for embryo-transfer were used in this study. Donated embryos were thawed and processed, and then their morphological behavior during the initiation of compaction was dynamically observed via time-lapse cinematography (TLC) for 5 days. Although the initiation of compaction occurred throughout the period from the 4-cell to 16-cell stage, 99 (86.1 %) embryos initiated compaction at the 8-cell stage or later, with initiation at the 8-cell stage being most frequent (22.6 %). Of these 99 embryos, 49.5 % developed into good-quality blastocysts. In contrast, of the 16 (13.9 %) embryos that initiated compaction prior to the 8-cell stage, only 18.8 % developed into good-quality blastocysts. Embryos that initiated compaction before the 8-cell stage showed significantly higher numbers of multinucleated blastomeres, due to asynchronism in nuclear division at the third mitotic division resulting from cytokinetic failure. The initiation of compaction primarily occurs at the third mitotic division or later in human embryos. Embryos that initiate compaction before the 8-cell stage are usually associated with aberrant embryonic development (i.e., cytokinetic failure accompanied by karyokinesis).

  8. Maintenance ofMestimprinted methylation in blastocyst-stage mouse embryos is less stable than other imprinted loci following superovulation or embryo culture.

    Science.gov (United States)

    Velker, Brenna A M; Denomme, Michelle M; Krafty, Robert T; Mann, Mellissa R W

    2017-07-01

    Assisted reproductive technologies are fertility treatments used by subfertile couples to conceive their biological child. Although generally considered safe, these pregnancies have been linked to genomic imprinting disorders, including Beckwith-Wiedemann and Silver-Russell Syndromes. Silver-Russell Syndrome is a growth disorder characterized by pre- and post-natal growth retardation. The Mest imprinted domain is one candidate region on chromosome 7 implicated in Silver-Russell Syndrome. We have previously shown that maintenance of imprinted methylation was disrupted by superovulation or embryo culture during pre-implantation mouse development. For superovulation, this disruption did not originate in oogenesis as a methylation acquisition defect. However, in comparison to other genes, Mest exhibits late methylation acquisition kinetics, possibly making Mest more vulnerable to perturbation by environmental insult. In this study, we present a comprehensive evaluation of the effects of superovulation and in vitro culture on genomic imprinting at the Mest gene. Superovulation resulted in disruption of imprinted methylation at the maternal Mest allele in blastocysts with an equal frequency of embryos having methylation errors following low or high hormone treatment. This disruption was not due to a failure of imprinted methylation acquisition at Mest in oocytes. For cultured embryos, both the Fast and Slow culture groups experienced a significant loss of maternal Mest methylation compared to in vivo -derived controls. This loss of methylation was independent of development rates in culture. These results indicate that Mest is more susceptible to imprinted methylation maintenance errors compared to other imprinted genes.

  9. Long term costs and effects of reducing the number of twin pregnancies in IVF by single embryo transfer: the TwinSing study

    Directory of Open Access Journals (Sweden)

    van Goudoever Johannes B

    2010-10-01

    Full Text Available Abstract Background Pregnancies induced by in vitro fertilisation (IVF often result in twin gestations, which are associated with both maternal and perinatal complications. An effective way to reduce the number of IVF twin pregnancies is to decrease the number of embryos transferred from two to one. The interpretation of current studies is limited because they used live birth as outcome measure and because they applied limited time horizons. So far, research on long-term outcomes of IVF twins and singletons is scarce and inconclusive. The objective of this study is to investigate the short (1-year and long-term (5 and 18-year costs and health outcomes of IVF singleton and twin children and to consider these in estimating the cost-effectiveness of single embryo transfer compared with double embryo transfer, from a societal and a healthcare perspective. Methods/Design A multi-centre cohort study will be performed, in which IVF singletons and IVF twin children born between 2003 and 2005 of whom parents received IVF treatment in one of the five participating Dutch IVF centres, will be compared. Data collection will focus on children at risk of health problems and children in whom health problems actually occurred. First year of life data will be collected in approximately 1,278 children (619 singletons and 659 twin children. Data up to the fifth year of life will be collected in approximately 488 children (200 singletons and 288 twin children. Outcome measures are health status, health-related quality of life and costs. Data will be obtained from hospital information systems, a parent questionnaire and existing registries. Furthermore, a prognostic model will be developed that reflects the short and long-term costs and health outcomes of IVF singleton and twin children. This model will be linked to a Markov model of the short-term cost-effectiveness of single embryo transfer strategies versus double embryo transfer strategies to enable the

  10. Determining the status of non-transferred embryos in Ireland: a conspectus of case law and implications for clinical IVF practice.

    LENUS (Irish Health Repository)

    Sills, Eric Scott

    2009-01-01

    The development of in vitro fertilisation (IVF) as a treatment for human infertilty was among the most controversial medical achievements of the modern era. In Ireland, the fate and status of supranumary (non-transferred) embryos derived from IVF brings challenges both for clinical practice and public health policy because there is no judicial or legislative framework in place to address the medical, scientific, or ethical uncertainties. Complex legal issues exist regarding informed consent and ownership of embryos, particularly the use of non-transferred embryos if a couple separates or divorces. But since case law is only beginning to emerge from outside Ireland and because legislation on IVF and human embryo status is entirely absent here, this matter is poised to raise contractual, constitutional and property law issues at the highest level. Our analysis examines this medico-legal challenge in an Irish context, and summarises key decisions on this issue rendered from other jurisdictions. The contractual issues raised by the Roche case regarding informed consent and the implications the initial judgment may have for future disputes over embryos are also discussed. Our research also considers a putative Constitutional \\'right to procreate\\' and the implications EU law may have for an Irish case concerning the fate of frozen embryos. Since current Medical Council guidelines are insufficient to ensure appropriate regulation of the advanced reproductive technologies in Ireland, the report of the Commission on Assisted Human Reproduction is most likely to influence embryo custody disputes. Public policy requires the establishment and implementation of a more comprehensive legislative framework within which assisted reproductive medical services are offered.

  11. Determining the status of non-transferred embryos in Ireland: a conspectus of case law and implications for clinical IVF practice.

    Science.gov (United States)

    Sills, Eric Scott; Murphy, Sarah Ellen

    2009-07-09

    The development of in vitro fertilisation (IVF) as a treatment for human infertilty was among the most controversial medical achievements of the modern era. In Ireland, the fate and status of supranumary (non-transferred) embryos derived from IVF brings challenges both for clinical practice and public health policy because there is no judicial or legislative framework in place to address the medical, scientific, or ethical uncertainties. Complex legal issues exist regarding informed consent and ownership of embryos, particularly the use of non-transferred embryos if a couple separates or divorces. But since case law is only beginning to emerge from outside Ireland and because legislation on IVF and human embryo status is entirely absent here, this matter is poised to raise contractual, constitutional and property law issues at the highest level. Our analysis examines this medico-legal challenge in an Irish context, and summarises key decisions on this issue rendered from other jurisdictions. The contractual issues raised by the Roche case regarding informed consent and the implications the initial judgment may have for future disputes over embryos are also discussed. Our research also considers a putative Constitutional 'right to procreate' and the implications EU law may have for an Irish case concerning the fate of frozen embryos. Since current Medical Council guidelines are insufficient to ensure appropriate regulation of the advanced reproductive technologies in Ireland, the report of the Commission on Assisted Human Reproduction is most likely to influence embryo custody disputes. Public policy requires the establishment and implementation of a more comprehensive legislative framework within which assisted reproductive medical services are offered.

  12. Determining the status of non-transferred embryos in Ireland: a conspectus of case law and implications for clinical IVF practice

    Directory of Open Access Journals (Sweden)

    Sills Eric

    2009-07-01

    Full Text Available Abstract The development of in vitro fertilisation (IVF as a treatment for human infertilty was among the most controversial medical achievements of the modern era. In Ireland, the fate and status of supranumary (non-transferred embryos derived from IVF brings challenges both for clinical practice and public health policy because there is no judicial or legislative framework in place to address the medical, scientific, or ethical uncertainties. Complex legal issues exist regarding informed consent and ownership of embryos, particularly the use of non-transferred embryos if a couple separates or divorces. But since case law is only beginning to emerge from outside Ireland and because legislation on IVF and human embryo status is entirely absent here, this matter is poised to raise contractual, constitutional and property law issues at the highest level. Our analysis examines this medico-legal challenge in an Irish context, and summarises key decisions on this issue rendered from other jurisdictions. The contractual issues raised by the Roche case regarding informed consent and the implications the initial judgment may have for future disputes over embryos are also discussed. Our research also considers a putative Constitutional 'right to procreate' and the implications EU law may have for an Irish case concerning the fate of frozen embryos. Since current Medical Council guidelines are insufficient to ensure appropriate regulation of the advanced reproductive technologies in Ireland, the report of the Commission on Assisted Human Reproduction is most likely to influence embryo custody disputes. Public policy requires the establishment and implementation of a more comprehensive legislative framework within which assisted reproductive medical services are offered.

  13. The nasal apparatus of the red squirrel (Sciurus vulgaris L.) embryo at the stage of the fully formed chondrocranium.

    Science.gov (United States)

    Slabý, O

    1991-01-01

    The nasal apparatus of the squirrel embryo at the optimum stage of the chondrocranium displays simple (though not always primitive) features and individual structures developed largely as in other rodents and even in insectivores. Primitive features include the presence of a cartilago paraseptalis communis and probably the simplicity of the olfactory labyrinth, whose main support in the region as a whole is ethmoturbinale I, whose dorsal and ventral lamella divide off the basic recesses; anterior (frontalis), maxillaris and frontoturbinalis. Completely caudally we find ethmoturbinale I, the frontoturbinalia and the relevant secondary (greatly reduced) recesses. The zona annularis is interrupted by failure of the rostral processes of the lamina transversalis anterior (corresponding to the processes laterales ventrales) to fuse with the capsula nasi ant. or with the anlage of the septum nasi. We did find a fenestra nasi superior (lateralis). The atrioturbinale is well developed, the maxilloturbinale only as a trace, but we have a very striking nasoturbinale. In the rostral region of the olfactory segment, the cartilaginous capsule has a very conspicuous thickening, which in this part is formed of paranasal cartilage. The thickening presents outwardly as the prominentia anterior; cavity of the recessus anterior (frontalis) is formed inside it. The rest of the dorsal and caudal wall of this cavity forms the crista semicircularis, which further caudally circumscribes the foramen olfactorium. A foramen epiphaniale is present. The vestibular region of the epithelial nasal tube is interestingly formed. In cross section it is crescent-shaped; the nasal tube itself opens into the convexity, but the lamina transversalis ant. sends a turbinale into the concavity. A cartilago alaris superior, which develops independently in situ, is present; in our stage it is associated with the wall of the capsule, but ventrolaterally it terminates freely and is not joined to any other

  14. Characterization of secreted proteins of 2-cell mouse embryos cultured in vitro to the blastocyst stage with and without protein supplementation.

    Science.gov (United States)

    Burch, Tanya; Yu, Liang; Nyalwidhe, Julius; Horcajadas, Jose A; Bocca, Silvina; Swanson, R James; Oehninger, Sergio

    2014-06-01

    To identify the secreted proteins of murine embryos grown in vitro. Two-cell mouse embryos (n=432) were randomly allocated to culture to the blastocyst stage in protein-free and in protein-supplemented (3 % BSA) media. Proteins were separated by SDS-PAGE; bands were visualized by coomassie staining, followed by in-gel trypsin digestion and liquid chromatography-tandem mass spectrometry. RT-PCR and confocal microscopy were used to confirm gene/protein expression in blastocysts. Of all individually identified proteins, 34 and 23 were found in embryos cultured without and with BSA, respectively, and 20 were common. Identified proteins having an N-terminal secretory sequence or transmembrane domains located on the extracellular backbone were postulated as secreted proteins. Gene and protein expression for two selected molecules were confirmed. Functional analysis revealed over-represented processes related to lipid metabolism, cyclase activity, and cell adhesion/membrane functions. This study provided evidence to further characterize secreted proteins by mouse embryos grown from the 2-cell to the blastocyst stage in vitro. Because of homology between murine and human, these results may provide information to be translated to the clinical setting.

  15. Embryos, microscopes, and society.

    Science.gov (United States)

    Maienschein, Jane

    2016-06-01

    Embryos have different meanings for different people and in different contexts. Seen under the microscope, the biological embryo starts out as one cell and then becomes a bunch of cells. Gradually these divide and differentiate to make up the embryo, which in humans becomes a fetus at eight weeks, and then eventually a baby. At least, that happens in those cases that carry through normally and successfully. Yet a popular public perception imagines the embryo as already a little person in the very earliest stages of development, as if it were predictably to become an adult. In actuality, cells can combine, pull apart, and recombine in a variety of ways and still produce embryos, whereas most embryos never develop into adults at all. Biological embryos and popular imaginations of embryos diverge. This paper looks at some of the historical reasons for and social implications of that divergence. Copyright © 2016 The Author. Published by Elsevier Ltd.. All rights reserved.

  16. Stepwise in-straw dilution and direct transfer using open pulled straws (OPS) in the mouse: a potential model for field manipulation of vitrified embryos.

    Science.gov (United States)

    Yang, Qi-En; Hou, Yun-Peng; Zhou, Guang-Bin; Yang, Zhong-Qiang; Zhu, Shi-En

    2007-04-01

    In the present study, mouse blastocysts were employed to investigate the feasibility and efficiency of stepwise in-straw dilution and direct transfer using the open pulled straw (OPS) method. In experiment I, the effects of various vitrification solutions (VS) on embryo survival were examined. After thawing, the expanded blastocyst rates (97.59 and 95.05%) and hatching rates (80.48 and 78.95%) achieved in the EDFS30 [15% ethylene glycol (EG), 15% dimethyl sulfoxide (DMSO), Ficoll, and sucrose] and EFS40 [40% EG, Ficoll, and sucrose] groups were no different from those (96.15% and 83.33%) of the control group. However, the rates in the EFS30 [30% EG, Ficoll, and sucrose] (87.80 and 55.43%) and EDFS40 [20% EG, 20% DMSO, Ficoll, and sucrose] (95.69 and 70.97%) groups were significantly lower than those (96.15 and 83.33%) of the control group (Pcontrol groups. The embryos could be successfully thawed by immersing the OPS in 0.5 M sucrose for 3 min and then 0.25 M sucrose for 2 min. In experiment III, the effect of immersion time of the OPS in diluent (PBS) on the viability of vitrified embryos was investigated. After in-straw thawing, OPSs were immersed immediately in 1 ml PBS for 0 to 30 min. When the immersion time of the OPSs in PBS was less than 12 min, in vitro development of the in-straw thawed embryos was no different from that of the controls. In experiment IV, in-straw thawed blastocysts were directly transferred to pseudopregnant mice to examine their in vivo developmental viability. The pregnancy (91.67%) and birth rates (42.42%) of embryos in-straw thawed and directly transferred were no different from those of the unvitrified controls (90.90 and 40%) and embryos thawed by the conventional method (84.61 and 46.94%). These results demonstrate that mouse embryos vitrified with OPS could be successfully thawed by stepwise in-straw dilution and transferred directly to a recipient and that this method might be a model for field manipulation of vitrified embryos

  17. High in vitro development after somatic cell nuclear transfer and trichostatin A treatment of reconstructed porcine embryos

    DEFF Research Database (Denmark)

    Li, J.; Østrup, Olga; Villemoes, Klaus

    2008-01-01

    Abnormal epigenetic modification is supposed to be one of factors accounting for inefficient reprogramming of the donor cell nuclei in ooplasm after somatic cell nuclear transfer (SCNT). Trichostatin A (TSA) is an inhibitor of histone deacetylase, potentially enhancing cloning efficiency. The aim...... of our present study was to establish the optimal TSA treatment in order to improve the development of handmade cloned (HMC) porcine embryos and examine the effect of TSA on their development. The blastocyst percentage of HMC embryos treated with 37.5 nM TSA for 22-24 h after activation increased up...

  18. Lapse in embryo transfer training does not negatively affect clinical pregnancy rates for reproductive endocrinology and infertility fellows.

    Science.gov (United States)

    Kresowik, Jessica; Sparks, Amy; Duran, Eyup H; Shah, Divya K

    2015-03-01

    To compare rates of clinical pregnancy (CPR) and live birth (LBR) following embryo transfer (ET) performed by reproductive endocrinology and infertility (REI) fellows before and after a prolonged lapse in clinical training due to an 18-month research rotation. Retrospective cohort study. Not applicable. All women undergoing in vitro fertilization (IVF) and IVF-intracytoplasmic sperm injection (ICSI) cycles with ET performed by REI fellows from August 2003 to July 2012. Eighteen-month lapse in clinical training of REI fellows. CPR and LBR before and after the lapse in clinical training were calculated and compared per fellow and as a composite group. Alternating logistic regression models were used to calculate the odds of clinical pregnancy and live birth following transfers performed before and after the lapse in training. Unadjusted odds of clinical pregnancy and live birth were similar between the two time periods both for individual fellows and for the composite group. Alternate logistic regression analysis revealed no significant difference in CPR (odds ratio [OR] 0.94, 95% confidence interval [CI] 0.83-1.07) or LBR (OR 1.05, 95% CI 0.94-1.18) after the lapse in training compared with before. A research rotation is common in REI fellowship training programs. This prolonged departure from clinical training does not appear to negatively affect pregnancy outcome following fellow ET. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  19. Accuracy of transferring microparts in a multi stage former

    DEFF Research Database (Denmark)

    Mahshid, Rasoul; Hansen, Hans Nørgaard; Arentoft, Mogens

    2013-01-01

    and the second one is intended to depict how the unit transfers the parts with different diameters with respect to the front profile of the fingers. The experiments showed that the manipulator can handle the parts with 7 μm accuracy, 2 μm reproducibility and 9μm uncertainty for a 20mm distance between two...

  20. Application of in vitro production-embryo transfer in the protection ...

    African Journals Online (AJOL)

    Yomi

    2012-01-12

    Jan 12, 2012 ... After coordination with two industrial husbandry complexes, superior cows exposed to cull were transferred to the slaughter- house. Ovaries of superior cow were collected at abattoirs immediately after slaughter and transported to the laboratory in saline solution (0.9% NaCl) supplemented with penicillin G ...

  1. Embryo-fetal transfer of bevacizumab (Avastin) in the rat over the course of gestation and the impact of neonatal Fc receptor (FcRn) binding.

    Science.gov (United States)

    Thorn, Mitchell; Piche-Nicholas, Nicole; Stedman, Donald; Davenport, Scott W; Zhang, Ning; Collinge, Mark; Bowman, Christopher J

    2012-10-01

    There is concern about embryo-fetal exposure to antibody-based biopharmaceuticals based on the increase of such therapies being prescribed to women of childbearing potential. Therefore, there is a desire to better characterize embryo-fetal exposure of these molecules. The pregnant rat is a standard model for evaluating the potential consequences of exposure but placental transfer of antibody-based biopharmaceuticals is not well understood in this model. The relative embryo-fetal distribution of an antibody-based biopharmaceutical was evaluated in the rat. Bevacizumab (Avastin) was chosen as a tool antibody since it does not have significant target binding in the rat that might influence embryo-fetal biodistribution. Avastin was labeled with a fluorescent dye, characterized, and injected into pregnant rats at different gestation ages. Labeled Avastin in fetal tissues was visualized ex vivo using an IVIS 200 (Caliper, A PerkinElmer Company, Alameda, CA). Avastin localized to the fetus as early as 24-hr post intravenous injection of the dam, and was taken up by the fetus in a dose-dependent manner. Avastin was detectable in the developing embryo as early as gestation day 13 and continued to be transferred until the end of gestation. Fetal transfer of Avastins mutated in the portion of the antibody that binds the neonatal Fc receptor (FcRn) was tested in late gestation and was found to correlate with affinities of the mutant Avastin antibody to FcRn. The novel application of this imaging technology was used to characterize the onset and duration of Avastin maternal-fetal transfer in rats and the importance of FcRn binding. © 2012 Wiley Periodicals, Inc.

  2. Preimplantation development of embryos in women of advanced maternal age

    Directory of Open Access Journals (Sweden)

    O. V. Chaplia

    2014-04-01

    Full Text Available In order to reveal the influence of genetic component on the early embryo development, the retrospective study of morphokinetic characteristics of 717 embryos subjected to preimplantation genetic testing was conducted. Blastomere biopsy for FISH-based preimplantation genetic screening of 7 chromosomes was performed on the third day of culture, while embryo developmental potential and morphological features at the cleavage and blastulation stage were studied regarding maternal age particularly in the group of younger women and patients older than 36. Results of genetic testing revealed that euploid embryos rate gradually decreased with maternal age comprising 39.9% in young women group and 25.3% of specimen belonging to elder patients. At the cleavage stage, morphological characteristics of aneuploid and euploid embryos didn’t differ significantly regardless of the age of patients that could be accounted for the transcriptional silence of embryo genome till the third day of its development. However, in case of prolonged culture chromosomally balanced embryos rarely faced developmental arrest (in 7.9% and formed blastocysts half more frequently compared to aberrant embryos (respectively 75.6 versus 49.8%. Nevertheless, no substantial difference was found between blastocyst formation rate among embryos with similar genetic component regardless of the maternal age. Taking into consideration high rate of chromosomally unbalanced embryos specific to patients of advanced maternal age, the relative proportion of aneuplouid blastocysts was significantly higher in this group of embryos. Thus, without genetic screening there is a possibility of inaccurate selection of embryos for women of advanced reproductive age for transfer procedure even in case of prolonged culture. Consequently, increase of aneuploid embryos frequency associated with permanent preimplantation natural selection effectiveness along with the postimplantation natural selection failure

  3. Acetylcholinesterase-positive innervation is present at undifferentiated stages of the sea turtle Lepidochelis olivacea embryo gonads: implications for temperature-dependent sex determination.

    Science.gov (United States)

    Gutiérrez-Ospina, G; Jiménez-Trejo, F J; Favila, R; Moreno-Mendoza, N A; Granados Rojas, L; Barrios, F A; Díaz-Cintra, S; Merchant-Larios, H

    1999-07-19

    In embryos of different reptile species, incubation temperature triggers a cascade of endocrine events that lead to gonad sex differentiation. The cellular and molecular mechanisms by which temperature sets in motion this process are still controversial. Here, we begin evaluating the possible participation of the nervous system in temperature-dependent sex determination by showing the existence and origin of acetylcholinesterase (AchE)-positive nerve fibers in undifferentiated gonads of the Lepidochelys olivacea (L. olivacea) sea turtle putative male and female embryos, along the thermosensitive period for sex determination (TPSD; stages 20-27). AChE-positive nerve bundles and fibers were readily visualized until developmental stage 24 and thereafter. DiI injections and confocal imaging showed that some of these gonadal nerves arise from the lower thoracic and upper lumbar spinal cord levels, and might thus be sensory in nature. Because the vertebrate spinal cord is capable of integrating by itself thermoregulatory responses with no intervention of uppermost levels of the central nervous system, we also evaluated spinal cord maturation during the TPSD. The maturation of the spinal cord was more advanced in putative female than in male embryos, when sex determination is taking place for each sex; this process starts and ends earlier in male than in female embryos. Together these observations open the possibility that the spinal cord and the innervation derived from it could play a direct role in driving or modulating the process of temperature-dependent gonad sex determination and/or differentiation, particularly in female L. olivacea embryos.

  4. Kebuntingan Hasil Transfer Blastosis Mencit yang Dibekukan dengan Metode Vitrifikasi Kriolup

    Directory of Open Access Journals (Sweden)

    I Wayan Batan

    2012-11-01

    Full Text Available The aim of the study was to assess the viability of vitrified embryo using cryoloop as a carrier ofembryo. The blastocyst stage embryos were collected from superovulated mice. Embryos were frozenusing vitrification method and vitrified embryos were loaded on copper filament cryoloop before dipped inliquid nitrogen. The viability of vitrified embryos was assess in vitro by medium cultered and in vivo bytransfered them to recipient mice. The result shows the viability of vitrified embryos was 85,7% after 24hours cultured and the embryos were born from two pregnant recipient mice out of nine (22% or fouroffspring out of 63 trasfered embryos (6%. In conclusion, vitrified blatocyst stage embryos using cryoloopas a carrier could keep the viability of the embryos and they could be transfered to the recipient mice andwere born normally.

  5. HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yingying [Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming 650223 (China); State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Hai, Tang; Liu, Zichuan; Zhou, Shuya; Lv, Zhuo [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Ding, Chenhui; Liu, Lei [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Niu, Yuyu [Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming 650223 (China); Zhao, Xiaoyang; Tong, Man [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Graduate University of Chinese Academy of Sciences, Beijing 100049 (China); Wang, Liu [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China); Jouneau, Alice [INRA, UMR 1198, ENVA, CNRS, FRE 2857, Biologie du Developpement et Reproduction, Jouy en Josas F-78350 (France); Zhang, Xun [Neuroendocrine Unit, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 (United States); Ji, Weizhi, E-mail: wji@mail.kiz.ac.cn [Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming 650223 (China); Zhou, Qi, E-mail: qzhou@ioz.ac.cn [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China)

    2010-07-02

    Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo.

  6. HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

    International Nuclear Information System (INIS)

    Wang, Yingying; Hai, Tang; Liu, Zichuan; Zhou, Shuya; Lv, Zhuo; Ding, Chenhui; Liu, Lei; Niu, Yuyu; Zhao, Xiaoyang; Tong, Man; Wang, Liu; Jouneau, Alice; Zhang, Xun; Ji, Weizhi; Zhou, Qi

    2010-01-01

    Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo.

  7. Effect of Different Concentrations of Melatonin on Live Births Resulting from the Transfer of Two-Cell Embryos of NMRI Mice

    Directory of Open Access Journals (Sweden)

    Mahdi Saadati

    2014-12-01

    Full Text Available Background & objectives : Infertility is a global problem affecting millions of men and women in developed and developing countries. In this regard, in-vitro fertilization (IVF plays an important role in improving the quality of life in infertile patients. However, studies have shown that the implantation failure in IVF is the main challenge of this procedure. Melatonin can increase the survival rate of embryos and IVF success rate through eliminating free radicals and removing reactive oxygen species. So, this study is conducted to investigate the effects of different concentrations of melatonin on the rate of newborns of mice following transfer oftwo-cell embryos .   Methods : In this study, female mice with average age of six to eight weeks were superovulated by administering pregnant mares serum gonadotropin (PMSG intraperitoneally (7.5 IU. ip, and followed after 48h by human chorionic gonadotropin (hCG (7.5 IU. ip. Two-cell mouse embryos were obtained from female mice oviduct after 48 h. The embryos transferred bilaterally into pseudopregnant mice of the same strain through surgical procedure and 8-14 embryos were transferred to each tube. The study included 4 treatment groups and one control group (6 mice in each group. The treatment groups were exposed to subcutaneous injection of concentrations of 100 µm , 10 µm , 1 µm and 100 nm of melatonin. After the cesarean on 18th day of pregnancy, the percentage of live births was assessed. The outcomes of the live birth rate were as­sessed using the chi-square test and statistical analyses were carried out using SPSS version 16.0. Percentage of live birth was calculated and compared with the control group.   Results: A total of 701 two-cell mouse embryos were transferred into one control group and four experimental groups. The number and percentage of live births at concentrations of 100 µm and 10 µm of melatonin and the control groups were 21 (15.55%, 13 (9.15% and 9 (6

  8. Patient and cycle characteristics predicting high pregnancy rates with single-embryo transfer: an analysis of the Society for Assisted Reproductive Technology outcomes between 2004 and 2013.

    Science.gov (United States)

    Mersereau, Jennifer; Stanhiser, Jamie; Coddington, Charles; Jones, Tiffany; Luke, Barbara; Brown, Morton B

    2017-11-01

    To analyze factors associated with high live birth rate and low multiple birth rate in fresh and frozen-thawed assisted reproductive technology (ART) cycles. Retrospective cohort analysis. Not applicable. The study population included 181,523 women undergoing in vitro fertilization with autologous fresh first cycles, 27,033 with fresh first oocyte donor cycles, 37,658 with fresh second cycles, and 35,446 with frozen-thawed second cycles. None. Live birth rate and multiple birth rate after single-embryo transfer (SET) and double embryo transfer (DET) were measured, in addition to cycle characteristics. In patients with favorable prognostic factors, including younger maternal age, transfer of a blastocyst, and additional embryos cryopreserved, the gain in the live birth rate from SET to DET was approximately 10%-15%; however, the multiple birth rate increased from approximately 2% to greater than 49% in both autologous and donor fresh and frozen-thawed transfer cycles. This study reports a 10%-15% reduction in live birth rate and a 47% decrement in multiple birth rate with SET compared with DET in the setting of favorable patient prognostic factors. Our findings present an opportunity to increase the rate of SET across the United States and thereby reduce the multiple birth rate and its associated poor perinatal outcomes with assisted reproductive technology pregnancies. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  9. Effects of electric field strengths on fusion and in vitro development of domestic cat embryos derived by somatic cell nuclear transfer.

    Science.gov (United States)

    Karja, Ni Wayan Kurniani; Otoi, Takeshige; Wongsrikeao, Pimprapar; Shimizu, Ryohei; Murakami, Masako; Agung, Budiyanto; Fahrudin, Mokhamad; Nagai, Takashi

    2006-09-15

    The present study was conducted to determine the effect of electric field strength on the rate of membrane fusion between the somatic cell and cytoplast and on subsequent in vitro development of reconstructed embryos. Additionally, the in vitro developmental competence of cat oocytes artificially activated after 44 h of maturation culture was examined. An efficient fusion rate (64.2%) was obtained by applying a single pulse of 1.5 kV/cm for 50 micros, and the fusion rate remained almost constant at the higher field intensity (59.8 and 54.9% at 1.7 and 2.0 kV/cm, respectively). Although the cleavage rate of fused embryos increased with an increase of the electric field strength, there were no differences among the groups with respect to the proportion of development to the morula and blastocyst stages. In the additional experiment, oocytes at the metaphase II stage after culture for 44 h were activated by the combination of calcium ionophore (CaI) with cycloheximide (CHX). Some (11.8%) of activated oocytes developed to the blastocyst stage. Results from this study indicated that electric field strength affects the rates of fusion and cleavage but has no significant effects on the development to the blastocyst stage of reconstructed embryos. Prolonged maturation culture of cat oocytes (up to 44 h) decreased their ability to develop to the blastocyst stage.

  10. Bovine conceptus of Bos indicus produced by somatic cell nuclear transfer and parthenogenesis present morphological variations since the blastocyst stage

    Directory of Open Access Journals (Sweden)

    F.D. Oliveira

    2015-12-01

    Full Text Available In cattle, embryo development is characterized by the appearance of two distinct cell layers, the trophectoderm and the inner cell mass. The latter will undergo differentiation to form the embryonic disc consisting of the epiblast and hypoblast. The aim of this study was to ultrastructurally characterize the bovine embryo from different in vitro production techniques, with emphasis on trophectoderm and inner cell mass cells. Bovine embryos on day 7 (conception = D1 of pregnancy, derived via in vitro production techniques, were fixed for light and transmission electron microscopy processing. Results suggested that embryos produced by nuclear transfer of somatic cells and parthenogenesis showed significant changes in macroscopic and microscopic structure. Size was reduced, and the inner cell mass had no defined shape. Furthermore, organelles responsible for the absorption processes, communication, growth, and cellular metabolism were fewer and had changes in shape, when compared to results in embryos produced by in vitrofertilization. We concluded that embryos produced by parthenogenesis and SCNT exhibit morphological differences when compared with IVF embryos, such as undeveloped blastocoel, poorly defined distribution of ICM, and morphological differences in organelles.

  11. The First Human Cloned Embryo.

    Science.gov (United States)

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  12. Effects of recipient oocyte age and interval from fusion to activation on development of buffalo (Bubalus bubalis) nuclear transfer embryos derived from fetal fibroblasts.

    Science.gov (United States)

    Lu, F; Jiang, J; Li, N; Zhang, S; Sun, H; Luo, C; Wei, Y; Shi, D

    2011-09-15

    The objective was to investigate the effect of recipient oocyte age and the interval from activation to fusion on developmental competence of buffalo nuclear transfer (NT) embryos. Buffalo oocytes matured in vitro for 22 h were enucleated by micromanipulation under the spindle view system, and a fetal fibroblast (pretreated with 0.1 μg/mL aphidicolin for 24 h, followed by culture for 48 h in 0.5% fetal bovine serum) was introduced into the enucleated oocyte, followed by electrofusion. Both oocytes and NT embryos were activated by exposure to 5 μM ionomycin for 5 min, followed by culture in 2 mM 6-dimethyl-aminopurine for 3 h. When oocytes matured in vitro for 28, 29, 30, 31, or 32 h were activated, more oocytes matured in vitro for 30 h developed into blastocysts in comparison with oocytes matured in vitro for 32 h (31.3 vs 19.9%, P fusion (P fusion. However, 3 of 16 recipients were pregnant following transfer of blastocysts developed from the NT embryos activated at 3 h after fusion, and two of these recipients maintained pregnancy to term. We concluded that the developmental potential of buffalo NT embryos was related to recipient oocyte age and the interval from fusion to activation. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. Developmental competence of HMC(TM) derived bovine cloned embryos obtained from somatic cell nuclear transfer of adult fibroblasts and granulosa cells.

    Science.gov (United States)

    Bhojwani, Sanjay; Vajta, Gabor; Callesen, Henrik; Roschlau, Knut; Kuwer, Andreas; Becker, Frank; Alm, Hannelore; Torner, Helmut; Kanitz, Wilhelm; Poehland, Ralf

    2005-08-01

    To enable us to handle a large number of oocytes at a given time and to have an increased throughput of cloned embryos, we attempted the Handmade cloning (HMC) technique, a zona-free method of bovine somatic cell nuclear transfer. Our objective was to study the developmental competence of the HMC derived embryos obtained using different types of somatic cells. A total of 6,874 cumulus-oocyte-complexes were used with either 7th or 11th passage fibroblasts (1st and 2nd groups, respectively), which were prepared from male animals, or granulosa cells (3rd group) as nuclei donors. The average cleavage rate was 65%, accompanied by a blastocyst rate of just 2% for the cleaved products and 5% for the >8-cell embryos, and there was no significant difference between the three groups. Out of 27 blastocysts recovered, 22 blastocysts were transferred to 22 recipients, resulting in two pregnancies. One pregnancy was lost after the fourth week while the other progressed to full term with the birth of a male calf. This first successful cloning of a male calf with the HMC technique in Europe indicates the successful adoption and establishment of this technique in our laboratory, and that this technique can be successful in producing viable embryos.

  14. Rabbit embryonic stem cell lines derived from fertilized, parthenogenetic or somatic cell nuclear transfer embryos

    International Nuclear Information System (INIS)

    Fang, Zhen F.; Gai, Hui; Huang, You Z.; Li, Shan G.; Chen, Xue J.; Shi, Jian J.; Wu, Li; Liu, Ailian; Xu, Ping; Sheng, Hui Z.

    2006-01-01

    Embryonic stem cells were isolated from rabbit blastocysts derived from fertilization (conventional rbES cells), parthenogenesis (pES cells) and nuclear transfer (ntES cells), and propagated in a serum-free culture system. Rabbit ES (rbES) cells proliferated for a prolonged time in an undifferentiated state and maintained a normal karyotype. These cells grew in a monolayer with a high nuclear/cytoplasm ratio and contained a high level of alkaline phosphate activity. In addition, rbES cells expressed the pluripotent marker Oct-4, as well as EBAF2, FGF4, TDGF1, but not antigens recognized by antibodies against SSEA-1, SSEA-3, SSEA-4, TRA-1-10 and TRA-1-81. All 3 types of ES cells formed embryoid bodies and generated teratoma that contained tissue types of all three germ layers. rbES cells exhibited a high cloning efficiency, were genetically modified readily and were used as nuclear donors to generate a viable rabbit through somatic cell nuclear transfer. In combination with genetic engineering, the ES cell technology should facilitate the creation of new rabbit lines

  15. The development of a successful non-university-based ambulatory in vitro fertilization/embryo transfer program: phase I.

    Science.gov (United States)

    Sher, G; Knutzen, V; Stratton, C J; Montakhab, M M; Allenson, S G; Mayville, J; Rubenstein, J A; Glass, M J; Bilach, S M

    1984-04-01

    Most of the current in vitro fertilization and embryo transfer (IVF-ET) programs are university-based. The establishment of a successful ambulatory IVF program in association with a busy, two-man general obstetrics-gynecologic practice is described. Seventy-one infertile couples were screened between February 1 and October 15, 1983. Forty-three couples were judged eligible for IVF-ET. Forty-three women underwent a single attempt at ET. The first 13 of these women underwent controlled ovarian hyperstimulation (COH) with 150 mg clomiphene citrate and human chorionic gonadotropin (hCG), and the remaining 30 underwent COH with human menopausal gonadotropin (hMG) and hCG. One of the 13 patients who underwent COH with clomiphene citrate conceived but subsequently miscarried early in the first trimester, for an 8% pregnancy rate. There were 12 pregnancies among the 30 patients who received hMG and hCG, for a 40% pregnancy rate. Only three of these pregnancies miscarried in the early first trimester, and three of the nine viable pregnancies are twin gestations. The possible factors responsible for the high pregnancy rate with IVF-ET, using COH with hMG and hCG, are discussed, and the feasibility of its performance in a well-controlled, non-university program is demonstrated.

  16. Transgenic expression of green fluorescent protein in caprine embryos produced through electroporation-aided sperm-mediated gene transfer.

    Science.gov (United States)

    Kumar Pramod, R; Kumar, Rakesh; Mitra, Abhijit

    2016-01-15

    Current methods of transgenic animal production are afflicted by low efficiency and high cost. Recently, the electroporation aided sperm-mediated gene transfer (SMGT) emerges as a promising alternative with variable success rate. Among the domestic animal species, the electroporation-aided SMGT is less investigated in goats, except a few reports in which attempts have been made using the auto-uptake method of SMGT. In this study, we report an optimized electroporation condition for SMGT of caprine sperm cells. Results of this study demonstrated that electroporation of caprine sperm cells at 300 V for 200 mS in TALP medium allowed the maximum uptake of foreign DNA with minimum adverse effects on the vital semen parameters viz., progressive motility, viability, and membrane and acrosome integrity. Further, DNA binding assay revealed DNA uptake by 81.3% sperm cells when 1.0 μg of DNA was used under optimum electroporation conditions as compared to 16.5% on simple incubation. The qPCR analysis showed four-fold more (Pexpression of the foreign gene. In conclusion, our results confirm that using optimized electroporation conditions, the caprine sperm cells can uptake foreign DNA effectively with minimum negative effect on the semen parameters and could produce transgenic embryos. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Coasting, embryo development and outcomes of blastocyst transfer: a case-control study.

    Science.gov (United States)

    Talebi Chahvar, Solmas; Zosmer, Ariel; Caragia, Alina; Balestrini, Simona; Sabatini, Luca; Tranquilli, Andrea Luigi; Al-Shawaf, Talha

    2014-08-01

    This study compared the effect on blastocyst development and clinical outcome of coasting in women at increased risk of moderate-severe ovarian hyperstimulation syndrome (OHSS; n=389) with a control group matched for age and basal FSH that did not undergo coasting (n=386) in IVF/intracytoplasmic sperm injection (ICSI) cycles. The main outcome measures were rate of blastocyst development and live birth. More cycles progressed to the blastocyst stage in the coasted group (n=169) compared with the control group (n=83; 43.4% versus 21.5%; P<0.001). The biochemical pregnancy, clinical pregnancy and live birth rates were similar (46.5% versus 42.0%; 40.6% versus 37.8%; 31.6% versus 30.1%). The duration of coasting up to 4 days did not affect progression to blastocyst stage. The multivariate model showed that coasting (OR 1.73, P=0.004) and the number of oocytes retrieved (OR 1.17, P=0.001) were positively correlated with blastocyst formation. Coasting, a measure to reduce the risk of OHSS, does not impair blastocyst development or clinical outcome. Coasting should remain an effective measure to prevent OHSS. Copyright © 2014 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  18. Transfer, loss and physical processing of water in hit-and-run collisions of planetary embryos

    Science.gov (United States)

    Burger, C.; Maindl, T. I.; Schäfer, C. M.

    2018-01-01

    Collisions between large, similar-sized bodies are believed to shape the final characteristics and composition of terrestrial planets. Their inventories of volatiles such as water are either delivered or at least significantly modified by such events. Besides the transition from accretion to erosion with increasing impact velocity, similar-sized collisions can also result in hit-and-run outcomes for sufficiently oblique impact angles and large enough projectile-to-target mass ratios. We study volatile transfer and loss focusing on hit-and-run encounters by means of smooth particle hydrodynamics simulations, including all main parameters: impact velocity, impact angle, mass ratio and also the total colliding mass. We find a broad range of overall water losses, up to 75% in the most energetic hit-and-run events, and confirm the much more severe consequences for the smaller body also for stripping of volatile layers. Transfer of water between projectile and target inventories is found to be mostly rather inefficient, and final water contents are dominated by pre-collision inventories reduced by impact losses, for similar pre-collision water mass fractions. Comparison with our numerical results shows that current collision outcome models are not accurate enough to reliably predict these composition changes in hit-and-run events. To also account for non-mechanical losses, we estimate the amount of collisionally vaporized water over a broad range of masses and find that these contributions are particularly important in collisions of ˜ Mars-sized bodies, with sufficiently high impact energies, but still relatively low gravity. Our results clearly indicate that the cumulative effect of several (hit-and-run) collisions can efficiently strip protoplanets of their volatile layers, especially the smaller body, as it might be common, e.g., for Earth-mass planets in systems with Super-Earths. An accurate model for stripping of volatiles that can be included in future planet

  19. Occurrence of Campylobacter in the genitals of teaser bulls maintained at an embryo transfer center

    Directory of Open Access Journals (Sweden)

    Modolo J.R.

    2000-01-01

    Full Text Available Em central de transferência de embriões, após os procedimentos de reconhecimento do cio em 37 vacas receptoras, através de quatro rufiões vasectomizados, observou-se que 83% delas apresentavam retorno ao cio e algum corrimento serofibrinoso. Nos exames bacteriológicos realizados nos lavados prepuciais dos rufiões foi isolado o Campylobacter fetus subsp. venerealis em todos, fato que, analisado associadamente com o retorno ao cio das vacas receptoras, é indicativo da ocorrência de campilobacteriose no plantel. Essa ocorrência demonstra a necessidade de medidas eficazes de planejamento de saúde animal, pela utilização de rufiões com desvio lateral do pênis. Uma vez impossibilitado o contato sexual, seria impedida a transmissão do agente durante o coito. Torna-se imperioso consignar que a prática da prevenção racional de enfermidades continua sendo o procedimento mais econômico para uma produtividade animal mais rentável.

  20. Equine cloning: in vitro and in vivo development of aggregated embryos.

    Science.gov (United States)

    Gambini, Andrés; Jarazo, Javier; Olivera, Ramiro; Salamone, Daniel F

    2012-07-01

    The production of cloned equine embryos remains highly inefficient. Embryo aggregation has not yet been tested in the equine, and it might represent an interesting strategy to improve embryo development. This study evaluated the effect of cloned embryo aggregation on in vitro and in vivo equine embryo development. Zona-free reconstructed embryos were individually cultured in microwells (nonaggregated group) or as 2- or 3-embryo aggregates (aggregated groups). For in vitro development, they were cultured until blastocyst stage and then either fixed for Oct-4 immunocytochemical staining or maintained in in vitro culture where blastocyst expansion was measured daily until Day 17 or the day on which they collapsed. For in vivo assays, Day 7-8 blastocysts were transferred to synchronized mares and resultant vesicles, and cloned embryos were measured by ultrasonography. Embryo aggregation improved blastocyst rates on a per well basis, and aggregation did not imply additional oocytes to obtain blastocysts. Embryo aggregation improved embryo quality, nevertheless it did not affect Day 8 and Day 16 blastocyst Oct-4 expression patterns. Equine cloned blastocysts expanded and increased their cell numbers when they were maintained in in vitro culture, describing a particular pattern of embryo growth that was unexpectedly independent of embryo aggregation, as all embryos reached similar size after Day 7. Early pregnancy rates were higher using blastocysts derived from aggregated embryos, and advanced pregnancies as live healthy foals also resulted from aggregated embryos. These results indicate that the strategy of aggregating embryos can improve their development, supporting the establishment of equine cloned pregnancies.

  1. Economic and genetic performance of various combinations of in vitro-produced embryo transfers and artificial insemination in a dairy herd.

    Science.gov (United States)

    Kaniyamattam, Karun; Block, Jeremy; Hansen, Peter J; De Vries, Albert

    2018-02-01

    The objective of this study was to find the optimal proportions of pregnancies from an in vitro-produced embryo transfer (IVP-ET) system and artificial insemination (AI) so that profitability is maximized over a range of prices for embryos and surplus dairy heifer calves. An existing stochastic, dynamic dairy model with genetic merits of 12 traits was adapted for scenarios where 0 to 100% of the eligible females in the herd were impregnated, in increments of 10%, using IVP-ET (ET0 to ET100, 11 scenarios). Oocytes were collected from the top donors selected for the trait lifetime net merit (NM$) and fertilized with sexed semen to produce IVP embryos. Due to their greater conception rates, first ranked were eligible heifer recipients based on lowest number of unsuccessful inseminations or embryo transfers, and then on age. Next, eligible cow recipients were ranked based on the greatest average estimated breeding values (EBV) of the traits cow conception rate and daughter pregnancy rate. Animals that were not recipients of IVP embryos received conventional semen through AI, except that the top 50% of heifers ranked for EBV of NM$ were inseminated with sexed semen for the first 2 AI. The economically optimal proportions of IVP-ET were determined using sensitivity analysis performed for 24 price sets involving 6 different selling prices of surplus dairy heifer calves at approximately 105 d of age and 4 different prices of IVP embryos. The model was run for 15 yr after the start of the IVP-ET program for each scenario. The mean ± standard error of true breeding values of NM$ of all cows in the herd in yr 15 was greater by $603 ± 2 per cow per year for ET100 when compared with ET0. The optimal proportion of IVP-ET ranged from ET100 (for surplus dairy heifer calves sold for ≥$300 along with an additional premium based on their EBV of NM$ and a ≤$100 embryo price) to as low as ET0 (surplus dairy heifer calves sold at $300 with a $200 embryo price). For the default

  2. Somatic symptoms, sleep disturbance and psychological distress among women undergoing oocyte pick-up and in vitro fertilisation-embryo transfer.

    Science.gov (United States)

    Lin, Ya-Hui; Chueh, Ke-Hsin; Lin, Jia-Ling

    2016-06-01

    This study investigated the relationship between somatic symptoms, sleep disturbance and psychological distress in women who underwent oocyte pick-up and in vitro fertilisation-embryo transfer. According to worldwide research, women receiving assisted reproductive technologies may suffer from somatic and psychological symptoms and even experience sleep disturbance. Apparently, the guilt of infecundity forces Asian women to conceal this scenario and delay the time at which they accept medical assistance and mental support. A longitudinal study. The subjects in this study were infertile female patients who received oocyte pick-up and in vitro fertilisation-embryo transfer therapies in a hospital in northern Taiwan. Data were collected via a structured questionnaire, including somatic symptoms, Pittsburgh Sleep Quality Index and a five-item brief symptom rating scale. Data were analysed using the McNemar's test, Wilcoxon Sign Rank and fully entered multiple regression with spss version 20.0 software. The mean age of 100 participants was 34·54 (SD = 3·94) years old. They experienced abdominal distention, breast engorgement, nausea, faintness, diarrhoea, sleep disturbance and psychological distress when they received in vitro fertilisation-embryo transfer; these results were apparently higher than those receiving oocyte pick-up. In addition, sleep disturbance was the most significant factor involved in psychological distress during oocyte pick-up and in vitro fertilisation-embryo transfer therapies. The most serious indicator of the women's psychological distress during oocyte pick-up and in vitro fertilisation-embryo transfer treatment is anxiety. Sleep disturbance was the most significant factor involved in the psychological distress of women having problems with conception. Assisted reproductive technologies nurses can assess women's psychological distress by caring for their sleep disturbance without directly exploring their mood state. Moreover, these

  3. Climatic factors affecting quantity and quality grade of in vivo derived embryos of cattle.

    Science.gov (United States)

    Chinchilla-Vargas, Josué; Jahnke, Marianna M; Dohlman, Tyler M; Rothschild, Max F; Gunn, Patrick J

    2018-02-17

    The present study investigated the effects of climatic variables on the quality grade and quantity of in vivo derived cattle embryos in the Midwestern United States. Climatic information included greatest and least daily temperature, average daily wind speed and average temperature-humidity index for each of the 765 records. The response variables included the number of ovarian structures, viable embryos, quality grade 1 embryos, quality grade 2 embryos, quality grade 3 embryos, freezable embryos (sum of quality grade 1 and quality grade 2 embryos), transferable embryos (sum of quality grade 1-3 embryos), degenerate embryos and unfertilized ova. Measures for variables among the breeds of donors and sires grouped by geographical origin were compared. A negative effect of greater temperatures during the early embryonic development stage tended (P climatic variables have significant effects on the in vivo production of cattle embryos and that wind speed should be considered in future analyses of factors affecting embryo quality. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Effects of single dose GnRH agonist as luteal support on pregnancy outcome in frozen-thawed embryo transfer cycles: an RCT

    Directory of Open Access Journals (Sweden)

    Robab Davar

    2015-08-01

    Full Text Available Background: There is no doubt that luteal phase support is essential to enhance the reproductive outcome in IVF cycles. In addition to progesterone and human chorionic gonadotropin, several studies have described GnRH agonists as luteal phase support to improve implantation rate, pregnancy rate and live birth rate, whereas other studies showed dissimilar conclusions. All of these studies have been done in fresh IVF cycles. Objective: To determine whether an additional GnRH agonist administered at the time of implantation for luteal phase support in frozen-thawed embryo transfer (FET improves the embryo developmental potential. Materials and Methods: This is a prospective controlled trial study in 200 FET cycles, patients were randomized on the day of embryo transfer into group 1 (n=100 to whom a single dose of GnRH agonist (0.1 mg triptorelin was administered three days after transfer and group 2 (n=100, who did not receive agonist. Both groups received daily vaginal progesterone suppositories plus estradiol valerate 6 mg daily. Primary outcome measure was clinical pregnancy rate. Secondary outcome measures were implantation rate, chemical, ongoing pregnancy rate and abortion rate. Results: A total of 200 FET cycles were analyzed. Demographic data and embryo quality were comparable between two groups. No statistically significant difference in clinical and ongoing pregnancy rates was observed between the two groups (26% versus 21%, p=0.40 and 21% versus 17%, p=0.37, respectively. Conclusion: Administration of a subcutaneous GnRH agonist at the time of implantation does not increase clinical or ongoing pregnancy.

  5. Characterization of membrane lipid fluidity in human embryo cells malignantly transfer med post 238Pu α irradiation

    International Nuclear Information System (INIS)

    Qi Zirong; Sun Ling; Liu Guolian; Shen Zhiyuan

    1992-01-01

    The membrane lipid fluidity of malignantly transformed human embryo cells following 238 Pu α particlce irradiation in vitro has been studied. The results indicate that the ontogenesis depends on irradiation dose (Gy) and the membrane lipid fluidity in malignantly transformed cells is higher than that in normal embryo cells. With the microviscosity (η) of cells plotted against the cell counts, the correlation coefficient (γ) is calculated to be between 0.9936 and 0.9999. Since the malignant transformation of irradiated embryo cells is manifested early on cell membrane lipid, the fluidity of membrane lipid can be used as an oncologic marker

  6. Comparison between lignocaine hydrochloride and ropivacaine hydrochloride as lumbosacral epidural anaesthetic agents in goats undergoing laparoscopy assisted embryo transfer

    Directory of Open Access Journals (Sweden)

    Anubhav Khajuria

    2014-10-01

    Full Text Available Goats (n=12 undergoing laparoscopy assisted embryo transfer were randomly allotted to two groups (I and II and injected lignocaine hydrochloride (4mg/kg or ropivacaine hydrochloride (1mg/kg at the lumbosacral epidural space. The animals were held with raised hind quarters for first three minutes following injection. Immediately after induction of regional anaesthesia, they were restrained in dorsal recumbency in Trendelenburg position in a cradle. Laparoscopy was performed after creating pneumoperitoneum using filtered room air. The mean (± S.E induction time in animals of group I was significantly shorter (5.33 ± 0.61 min than those belonging to group II (12.66 ±1.99 min. Complete analgesia developed throughout the hind quarters and abdomen for 30 min and 60 min in group I and II animal’s respectively. Unlike animals of group I, group II goats continued to show moderate analgesia for 180 minutes. The motor activity returned after a lapse of 130.00 ± 12.64 min and 405.00 ± 46.31 min respectively. Occasional vocalization and struggling was noticed in two goats one from each group irrespective of the surgical manipulations during laparoscopy. The rectal temperature and respiration rates showed only non-significant increase, but the heart rate values were significantly higher (P < 0.5 up to 150 min in animals of both the groups when compared to their baseline values. From this study, it was concluded that both anaesthetic agents produced satisfactory regional anaesthesia in goats undergoing laparoscopy. However, considering the very long delay in regaining the hind limb motor activity, the use of ropivacaine may not be recommended for this purpose. Supplementation of sedative/tranquilizer with lumbosacral epidural anaesthesia needs evaluation.

  7. Comparison between an exclusive in vitro-produced embryo transfer system and artificial insemination for genetic, technical, and financial herd performance.

    Science.gov (United States)

    Kaniyamattam, K; Block, J; Hansen, P J; De Vries, A

    2017-07-01

    The objective of this study was to implement an in vitro-produced embryo transfer (IVP-ET) system in an existing stochastic dynamic dairy simulation model with multitrait genetics to evaluate the genetic, technical, and financial performance of a dairy herd implementing an exclusive IVP-ET or artificial insemination (AI) system. In the AI system, sexed semen was used on the genetically best heifers only. In the IVP-ET system, all of the animals in the herd were impregnated with female sexed embryos created through in vitro fertilization of oocytes collected from animals of superior genetics for different traits of interest. Each donor was assumed to yield on average 4.25 transferable embryos per collection. The remaining animals in the herd were used as recipients and received either a fresh embryo or a frozen embryo when fresh embryos were not available. Selection of donors was random or based on the greatest estimated breeding value (EBV) of lifetime net merit (NM$), milk yield, or daughter pregnancy rate. For both the IVP-ET and AI systems, culling of surplus heifer calves not needed to replace culled cows was based on the lowest EBV for the same traits. A herd of 1,000 milking cows was simulated 15 yr over time after the start of the IVP-ET system. The default cost to produce and transfer 1 embryo was set at $165. Prices of fresh embryos at which an exclusive IVP-ET system financially breaks even with the comparable AI system in yr 15 and for an investment period of 15 yr were also estimated. More surplus heifer calves were sold from the IVP-ET systems than from the comparable AI systems. The surplus calves from the IVP-ET systems were also genetically superior to the surplus calves from the comparable AI systems, which might be reflected in their market value as a premium price. The most profitable scenario among the 4 IVP-ET scenarios in yr 15 was the one in which NM$ was maximized in the herd. This scenario had an additional profit of $8/cow compared with a

  8. Embryo splitting

    OpenAIRE

    Karl Illmensee; Mike Levanduski

    2010-01-01

    Mammalian embryo splitting has successfully been established in farm animals. Embryo splitting is safely and efficiently used for assisted reproduction in several livestock species. In the mouse, efficient embryo splitting as well as single blastomere cloning have been developed in this animal system. In nonhuman primates embryo splitting has resulted in several pregnancies. Human embryo splitting has been reported recently. Microsurgical embryo splitting under Institutional Review Board appr...

  9. Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.

    Directory of Open Access Journals (Sweden)

    Maria Jesús Cánepa

    Full Text Available Reproductive biotechnologies such as in vitro fertilization (IVF and somatic cell nuclear transfer (SCNT enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70, endoplasmic reticulum (ER stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5 and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3 in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART.

  10. Effect of the time interval between fusion and activation on epigenetic reprogramming and development of bovine somatic cell nuclear transfer embryos.

    Science.gov (United States)

    Liu, Jun; Wang, Yongsheng; Su, Jianmin; Wang, Lijun; Li, Ruizhe; Li, Qian; Wu, Yongyan; Hua, Song; Quan, Fusheng; Guo, Zekun; Zhang, Yong

    2013-04-01

    Previous studies have shown that the time interval between fusion and activation (FA interval) play an important role in nuclear remodeling and in vitro development of somatic cell nuclear transfer (SCNT) embryos. However, the effects of FA interval on the epigenetic reprogramming and in vivo developmental competence of SCNT embryos remain unknown. In the present study, the effects of different FA intervals (0 h, 2 h, and 4 h) on the epigenetic reprogramming and developmental competence of bovine SCNT embryos were assessed. The results demonstrated that H3 lysine 9 (H3K9ac) levels decreased rapidly after fusion in all three groups. H3K9ac was practically undetectable 2 h after fusion in the 2-h and 4-h FA interval groups. However, H3K9ac was still evidently detectable in the 0-h FA interval group. The H3K9ac levels increased 10 h after fusion in all three groups, but were higher in the 2-h and 4-h FA interval groups than that in the 0-h FA interval group. The methylation levels of the satellite I region in day-7 blastocysts derived from the 2-h or 4-h FA interval groups was similar to that of in vitro fertilization blastocysts and is significantly lower than that of the 0-h FA interval group. SCNT embryos derived from 2-h FA interval group showed higher developmental competence than those from the 0-h and 4-h FA interval groups in terms of cleavage rate, blastocyst formation rate, apoptosis index, and pregnancy and calving rates. Hence, the FA interval is an important factor influencing the epigenetic reprogramming and developmental competence of bovine SCNT embryos.

  11. The developmental stage of chicken embryos modulates the impact of in ovo olfactory stimulation on food preferences.

    Science.gov (United States)

    Bertin, Aline; Calandreau, Ludovic; Arnould, Cécile; Lévy, Frédéric

    2012-03-01

    Like mammals, bird embryos are capable of chemosensory learning, but the ontogeny of their feeding preferences has not been examined. We tested if the timing of stimulation in chicken embryos modulates the impact of in ovo olfactory stimulation on later food preferences. We exposed chicken embryos to an olfactory stimulus for a 4-day period in the middle or toward the end of the incubation period. The chicks were tested for their preference between foods with and without the olfactory stimulus in 3-min choice tests and on a 24-h time scale. Regardless of the type of food (familiar or novel) or the duration of the test, the control chicks not exposed to the olfactory stimulus consistently showed significant preferences for non-odorized foods. Chicks that were exposed in ovo to the olfactory stimulus did not show a preference for odorized or non-odorized foods. Only those chicks that were exposed to the olfactory stimulus toward the end of the incubation period differed from the controls and incorporated a higher proportion of odorized food into their diets on a 24-h time scale. This result indicates that olfactory stimulation at the end of embryonic development has a stronger impact on later feeding preferences. Our findings contribute to the growing pool of recent data appreciating the impact of olfactory signals on behavior regulation in avian species.

  12. Preimplantation development of cloned canine embryos recovered by hysterectomy or surgical uterine flushing and subsequent pregnancy outcomes.

    Science.gov (United States)

    Jeong, Yeon Woo; Kim, Joung Joo; Kim, Hyun Duk; Hwang, Kyu Chan; Hyun, Sang Hwan; Kim, Nam-Hyung; Jeung, Eui-Bae; Hwang, Woo Suk

    2016-11-01

    Dog cloning offers a substantial potential because of the advancements in assisted reproductive technology and development of the human disease model in line with the transgenic technique. However, little is known about the development of the canine cloned embryo during the preimplantation period. The aim of this study was to investigate the most efficient method and time for collecting cloned canine preimplantation embryos and to ascertain the developmental timeline of cloned canine embryos. Two hundred cloned embryos were created and transferred into 11 surrogates. The preimplantation stage cloned embryos were then collected on Days 7, 8, and 9 using an ovariohysterectomy or the Foley balloon catheter method. The recovery rate of reconstructed embryos was 63.6% and 60.6% for the ovariohysterectomy and Foley balloon catheter methods, respectively. Although significant differences were observed in the early developmental stages (one-cell and 16-cell stages), no significant difference was observed in the blastocyst stage. Significantly higher blastocyst rate was observed when the embryos were collected on Day 8 (11.4%) than on Day 7 (0.0%; P cloned embryos can develop to blastocysts by Day 8, and full-term pregnancy can be achieved after embryo transfer in canine. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Urochordate ascidians possess a single isoform of Aurora kinase that localizes to the midbody via TPX2 in eggs and cleavage stage embryos.

    Directory of Open Access Journals (Sweden)

    Celine Hebras

    Full Text Available Aurora kinases are key proteins found throughout the eukaryotes that control mitotic progression. Vertebrate Aurora-A and B kinases are thought to have evolved from a single Aurora-kinase isoform closest to that found in present day urochordates. In urochordate ascidians Aurora binds both TPX2 (a vertebrate AURKA partner and INCENP (a vertebrate AURKB partner and localizes to centrosomes and spindle microtubules as well as chromosomes and midbody during both meiosis and mitosis. Ascidian Aurora also displays this localization pattern during mitosis in echinoderms, strengthening the idea that non-vertebrate deuterostomes such as the urochordates and echinoderms possess a single form of Aurora kinase that has properties of vertebrate Aurora-kinase A and B. In the ascidian, TPX2 localizes to the centrosome and the spindle poles also as in vertebrates. However, we were surprised to find that TPX2 also localized strongly to the midbody in ascidian eggs and embryos. We thus examined more closely Aurora localization to the midbody by creating two separate point mutations of ascidian Aurora predicted to perturb binding to TPX2. Both forms of mutated Aurora behaved as predicted: neither localized to spindle poles where TPX2 is enriched. Interestingly, neither form of mutated Aurora localized to the midbody where TPX2 is also enriched, suggesting that ascidian Aurora midbody localization required TPX2 binding in ascidians. Functional analysis revealed that inhibition of Aurora kinase with a pharmacological inhibitor or with a dominant negative kinase dead form of Aurora caused cytokinesis failure and perturbed midbody formation during polar body extrusion. Our data support the view that vertebrate Aurora-A and B kinases evolved from a single non-vertebrate deuterostome ancestor. Moreover, since TPX2 localizes to the midbody in ascidian eggs and cleavage stage embryos it may be worthwhile re-assessing whether Aurora A kinase or TPX2 localize to the midbody

  14. Expression pattern of pluripotent markers in different embryonic developmental stages of buffalo (Bubalus bubalis) embryos and putative embryonic stem cells generated by parthenogenetic activation.

    Science.gov (United States)

    Singh, Karn P; Kaushik, Ramakant; Garg, Veena; Sharma, Ruchi; George, Aman; Singh, Manoj K; Manik, Radhey S; Palta, Prabhat; Singla, Suresh K; Chauhan, Manmohan S

    2012-12-01

    In this study, we describe the production of buffalo parthenogenetic blastocysts and subsequent isolation of parthenogenetic embryonic stem cell (PGESC)-like cells. PGESC colonies exhibited dome-shaped morphology and were clearly distinguishable from the feeder layer cells. Different stages of development of parthenogenetic embryos and derived embryonic stem cell (ESC)-like cells expressed key ESC-specific markers, including OCT-4, NANOG, SOX-2, FOXD3, REX-1, STAT-3, TELOMERASE, NUCLEOSTEMIN, and cMYC. Immunofluorescence-based studies revealed that the PGESCs were positive for surface-based pluripotent markers, viz., SSEA-3, SSEA-4, TRA 1-80, TRA 1-60, CD-9, and CD-90 and exhibited high alkaline phosphatase (ALP) activity. PGEC cell-like cells formed embryoid body (EB)-like structures in hanging drop cultures and when cultured for extended period of time spontaneously differentiated into derivatives of three embryonic germ layers as confirmed by RT-PCR for ectodermal (CYTOKERATIN8, NF-68), mesodermal (MSX1, BMP-4, ASA), and endodermal markers (AFP, HNF-4, GATA-4). Differentiation of PGESCs toward the neuronal lineage was successfully directed by supplementation of serum-containing media with retinoic acid. Our results indicate that the isolated ESC-like cells from parthenogenetic blastocyst hold properties of ESCs and express markers of pluripotency. The pluripotency markers were also expressed by early cleavage-stage of buffalo embryos.

  15. Is there a link between blastomere contact surfaces of day 3 embryos and live birth rate?

    Directory of Open Access Journals (Sweden)

    Paternot Goedele

    2012-09-01

    Full Text Available Abstract Background Cell-cell communication and adhesion are essential for the compaction process of early stage embryos. The aim of this study was to develop a non-invasive objective calculation system of embryo compaction in order to test the hypothesis that embryos with a larger mean contact surface result in a higher live birth rate compared to embryos with a lower mean contact surface. Methods Multilevel images of 474 embryos transferred on day 3 were evaluated by the Cellify software. This software calculates the contact surfaces between the blastomeres. The primary outcome of this study was live birth. An ideal range of contact surface was determined and the positive and negative predictive value, the sensitivity, the specificity and the area under the curve for this new characteristic were calculated. Results In total, 115 (24% transferred embryos resulted in a live birth. Selection of an embryo for transfer on its mean contact surface could predict live birth with a high sensitivity (80% and high negative predicting value (83% but with a low positive predictive value (27%, a low specificity (31% and low area under the ROC curve (0.56. The mean contact surface of embryos cultured in a single medium was significantly higher compared to the mean contact surface of embryos cultured in a sequential medium (p = 0.0003. Conclusions Neither the mean contact surface nor the number of contact surfaces of a day 3 embryo had an additional value in the prediction of live birth. The type of culture medium, however, had an impact on the contact surface of an embryo. Embryos cultured in a single medium had a significant larger contact surface compared to embryos cultured in the sequential medium.

  16. Effect of Methotrexate or Salpingectomy for Ectopic Pregnancy on Subsequent In Vitro Fertilization-Embryo Transfer Outcomes.

    Science.gov (United States)

    Pereira, Nigel; Gerber, Deanna; Gerber, Rachel S; Lekovich, Jovana P; Elias, Rony T; Spandorfer, Steven D; Rosenwaks, Zev

    2015-01-01

    To investigate the effect of methotrexate (MTX) or salpingectomy for ectopic pregnancy on the outcomes of subsequent in vitro fertilization (IVF)-embryo transfer (ET) cycles. Retrospective cohort study (Canadian Task Force Classification II-3). Academic center. All patients undergoing fresh IVF-ET between January 2004 and July 2013 after treatment of an ectopic pregnancy with MTX or salpingectomy in the preceding IVF-ET cycle were analyzed for potential inclusion. MTX or laparoscopic salpingectomy for an ectopic pregnancy followed by a subsequent IVF-ET cycle. A total of 144 patients with sonographically confirmed ectopic pregnancies were identified during the study period. Of these, 107 (74.3%) patients were treated with MTX and 37 (25.7%) were treated with laparoscopic salpingectomy. Eighty-eight patients (82.2%) in the MTX group and 22 patients (59.4%) patients in the salpingectomy group underwent a subsequent IVF-ET cycle. There were no significant differences in demographic data or baseline cycle characteristics between the 2 groups. No difference was observed in basal follicle-stimulating hormone (FSH) level before and after MTX or salpingectomy treatment. Indicators of ovarian responsiveness, including total days of stimulation, total dosage of gonadotropins, and number of mature oocytes before and after either treatment, were comparable in the 2 groups. The number of doses of MTX (1 vs > 1) did not correlate with changes in ovarian response. The pregnancy outcomes, specifically live birth, were equivalent in the 2 groups. Comparing post-MTX cycles and post-salpingectomy cycles, patients in the latter group required higher doses of gonadotropins (+705 IU vs +221.5 IU; p ectopic pregnancies with MTX or salpingectomy might not adversely affect ovarian reserve, ovarian responsiveness, or subsequent IVF cycle outcomes. However, in our study cohort, patients treated with MTX, those s treated with laparoscopic salpingectomy required higher gonadotropin doses in a

  17. The action of X-rays on electrophysiological characteristics in early stages of development of frog embryo

    International Nuclear Information System (INIS)

    Tevdoradze, V.V.; Nadarejshvili, K.Sh.

    1987-01-01

    Comparative investigation of dynamics of changes in the membrane potential (MP), input resistance (R in ) and the coefficient of electric coupling (K ec ) between blastomers in the process of the first divisions of irradiated and nonirradiated Rana ridibunda frog embryo cleavage is carried out. Microelectrophysiological equipment has been used. It is detected that 12 Gy radiation dose does not change cycling of changes and absolute MP and K ec while the amplitute of cyclic oscillations (R in ) decreases sharpy. Possible mechanisms of this effect and perspectives of investigation of certain general problems on radiation membranology are discussed

  18. PGF₂α levels in Day 8 blood plasma are increased by the presence of one or more embryos in the uterus.

    Science.gov (United States)

    Gomez, E; Martin, D; Carrocera, S; Muñoz, M

    2015-08-01

    In cattle, the detection of very early endometrial responses is considered to be hampered by the presence of only a single embryo. Therefore, we have previously developed a model of multiple embryo transfer to circumvent this hindrance. In this work, we analysed embryo-maternal interactions in the bovine uterus on day 8 of development while comparing the presence of multiple v. single embryos using embryo transfer and artificial insemination, respectively. Concentration of proteins (β-actin, NFkB, clusterin and immunoproteosome 20S β5i subunit-i20S), by western blot, and hexoses (glucose and fructose) were measured in paired samples of uterine fluid (UF) from the same animal with and without embryos in the uterus and were compared with UF obtained after artificial insemination. Prostaglandin (PG) F2 α and PGE2 concentrations were also analysed in blood plasma. The four proteins analysed and hexoses were unaffected by the presence of one or more embryos in the uterus. However, blood PGF2 α showed similar, significant increases with one or more embryos over cyclic animals; such changes were not observed in blood PGE2. Although multiple embryo transfer may appear to be non-physiological, we showed that the uterus, at the very early embryonic stages, does exhibit physiological reactions. Multiple embryo transfer can, therefore, be used for studies of very early embryo-maternal interactions in vivo in monotocous species.

  19. Effect of air bubbles localization and migration after embryo transfer on assisted reproductive technology outcome.

    Science.gov (United States)

    Fıçıcıoğlu, Cem; Özcan, Pınar; Koçer, Melis G; Yeşiladalı, Mert; Alagöz, Oya; Özkara, Gülçin; Tayyar, Ahter T; Altunok, Çiğdem

    2018-02-01

    To evaluate the effect of embryo flash position and movement of the air bubbles at 1 and 60 minutes after ET on clinical pregnancy rates (PRs). Prospective clinical trial. University fertility clinic. A total of 230 fresh ultrasound-guided ETs performed by a single physician (C.F.) at the IVF center of Yeditepe University Hospital between September 2016 and February 2017 were included. Transabdominal ultrasonographic guidance at ET. Clinical PRs. There was no significant difference in terms of clinical PRs between women with embryo flash located >15 mm and women with embryo flash located 15 mm from the fundus were 47% and 60%, respectively (P=.6). The clinical intrauterine PRs were 69.5%, 38.5%, and 19.1% in fundal, static, and cervical, respectively. The highest PR was in fundal when compared with others (PReproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. In vitro fertilization in Japan - early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology.

    Science.gov (United States)

    Suzuki, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world's first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe. In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine. IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from an infertile woman, fertilizing them in vitro, and transferring fertilized eggs into the patient's uterine cavity (Fig. 1). Since the first report of successful IVF-ET, numerous techniques related to ART, such as cryopreservation of oocytes and embryos, gamete intrafallopian transfer (GIFT), and microinsemination, have been developed and refined (Table 1). Herein we describe the history of basic research in IVF-ET that led to human applications, how the birth of the first IVF-ET baby was achieved in Japan, the current status of ART in Japan, issues related to ART, and future prospects for ART.

  1. Treatment of Abnormal Vaginal Microbiota before Frozen Embryo Transfer: Case-Report and Minireview to Discuss the Longitudinal Treatment Efficacy of Oral Clindamycin

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    Thor Haahr

    2017-06-01

    Full Text Available Abnormal vaginal microbiota (AVM or bacterial vaginosis (BV might negatively impact reproductive outcomes of in vitro fertilization (IVF. However, before randomized controlled trials are initiated to investigate cause and effect, it is necessary to establish the optimal treatment for AVM. Metronidazole seems ineffective to treat the biofilm in AVM; thus, clindamycin could be suggested as a relevant antibiotic agent for future intervention based studies. In the present case report, we present the first longitudinal follow-up of the vaginal microbiota with molecular methods during and after oral clindamycin treatment. Furthermore, we review the recent literature with the aim to discuss the optimal AVM treatment in a fertility setting. The patient was 40 years old suffering from unexplained secondary infertility. Prior to the present transfer cycle, she had had two failed IVF cycles. The tentative explanation of failed treatment was age-related aneuploidy. However, the patient asked for AVM diagnosis and she was subsequently diagnosed and treated successfully. Unfortunately, the patient did not achieve pregnancy after clindamycin treatment and two subsequent frozen embryo transfer cycles. Taken together, we report an excellent AVM treatment efficacy both short-term and long-term following oral clindamycin treatment. We discuss the potential impact on the vaginal microbiota of co-treatment with estrogen patches in the stimulated frozen embryo transfer cycle. Furthermore, we discuss future aspects of AVM treatment such as the potential impact of estrogen and live biotherapeutic products to positively modulate the microbiota of the reproductive tract.

  2. The influence of animals from embryo transfer on the genetic evaluation of growth in Simmental beef cattle by using multi-trait models

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    Rodrigo Reis Mota

    2013-01-01

    Full Text Available The weight records from Simmental beef cattle were used in a genetic evaluation of growth with or without the inclusion of animals obtained by embryo transfer. A multi-trait model in which embryo transfer individuals were excluded (MTM1 contained 29,510 records from 10,659 animals, while another model without exclusion of these animals (MTM2 contained 62,895 weight records from 23,160 animals. The weight records were adjusted for ages of 100, 205, 365, 450, 550 and 730 days. The (covariance components and genetic parameters were estimated by the restricted maximum likelihood method. The (covariance components were similar in both models, except for maternal permanent environment variance. Direct heritabilities (h²d in MTM1 were 0.04, 0.11, 0.20, 0.27, 0.31 and 0.42, while in MTM2 they were 0.11, 0.11, 0.17, 0.21, 0.22 and 0.26 for 100, 205, 365, 450, 550 and 730 days of age, respectively. Estimates of h²d in MTM1 were higher than in MTM2 for the weight at 365 days of age. Genetic correlations between weights in both models ranged from moderate to high, suggesting that these traits may be determined mainly by the same genes. Animals from embryo transfer may be included in the genetic evaluation of Simmental beef cattle in Brazil; this inclusion may provide potential gains in accuracy and genetic gains by reducing the interval between generations.

  3. Pregnancy outcome and live birth after IVF and ICSI according to embryo quality.

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    Fauque, Patricia; Léandri, Roger; Merlet, Françoise; Juillard, Jean-Claude; Epelboin, Sylvie; Guibert, Juliette; Jouannet, Pierre; Patrat, Catherine

    2007-05-01

    The pregnancy outcome and the chances of birth were assessed according to embryo quality after IVF or ICSI. The implantation rate (IR), the loss of gestational sacs rate (LGSR), and birth rate (BR) were determined according to the cleavage stage and the integrity of blastomeres after day-2 homogeneous embryo transfers (n = 1812). The LGSR was higher after transfers of 2-3-cell or 5-6-cell embryos and was significantly increased when more than 20% of the embryo volume was fragmented in 4-cell embryos. After transfers of 4-cell embryos without fragmentation, the BR was significantly higher than the BR after transfers of 4-cell embryos with 1-20% fragmentation (16.6% vs 13.1%). The difference was the consequence of a higher IR (20.4% vs 17.3%) but also of a lower LGSR (18.9% vs 24.2%). Not only implantation and the ability to give a pregnancy, but also the capacity to give a live birth are dependent on the embryo quality.

  4. Effect of medium salt concentration on differentiation and maturation of somatic embryos of cassava (Manihot esculenta Crantz).

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    Groll, J; Mycock, D J; Gray, V M

    2002-05-01

    Culture of cassava somatic embryos on media with an altered macro- and micro-nutrient salt concentration affected embryo development and germination capability. In the tests, quarter-, half-, full- or double-strength Murashige and Skoog (MS) media were compared. The maximum number of somatic embryos differentiated from a proliferative nodular embryogenic callus (NEC) on either half- or full-strength MS medium, and the greatest numbers of cotyledonary stage embryos were formed on full-strength MS medium. Developed somatic embryos were then desiccated above a saturated K2SO4 solution for 10 d. After transfer to germination medium, embryos that had developed on half- and full-strength MS medium yielded 8.3 and 8.6 germinants g(-1) NEC tissue, respectively. For this important but often disregarded culture factor, either half- or full-strength MS medium is recommended for both the differentiation and development of cassava somatic embryos that are capable of germination.

  5. Risk and prevention of bovine viral diarrhea virus (BVDV) transmission through embryo production via somatic cell nuclear transfer (SCNT) using oocytes from persistently infected donors.

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    Gregg, K; Riddell, K P; Chen, S H; Galik, P K; Xiang, T; Guerra, T; Marley, M S; Polejaeva, I; Givens, M D

    2010-07-01

    The objective was to assess the risk of transmission of bovine viral diarrhea virus (BVDV) through embryo production via somatic cell nuclear transfer (SCNT), with oocytes obtained from persistently infected (PI) donors. Using ultrasound-guided follicular aspiration following superstimulation, oocytes were obtained from five female beef cattle, including three that were PI and two that were negative for BVDV. In the three PI cattle, seven aspirations yielded 32 oocytes (PI-1: three aspirations yielding six oocytes; PI-2: two aspirations yielding 14 oocytes; and PI-3: two aspirations yielding 12 oocytes). The oocyte recovery rate was better in negative control cattle, with 32 oocytes obtained from the two cattle in a single superstimulation and aspiration session. Oocytes were processed individually for SCNT, evaluated, and tested for BVDV. Nearly all (31/32) oocytes from the three PI donors were positive for BVDV by PCR, with detected viral RNA copy number ranging from 1 to 1.1 x 10(5). The proportion of oocytes acceptable for SCNT embryo production (based on oocyte quality and maturation status) was only 16 to 35% from PI donors, but was 81% from control donors. Therefore, routine testing of unacceptable (discarded) oocytes could be an effective approach to identify batches that might contain infected oocytes from PI donors. Identification and removal of high-risk batches of oocytes would minimize the risk of BVDV transmission through SCNT embryo production.

  6. Resurgence of Minimal Stimulation In Vitro Fertilization with A Protocol Consisting of Gonadotropin Releasing Hormone-Agonist Trigger and Vitrified-Thawed Embryo Transfer

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    Zhang John

    2016-07-01

    Full Text Available Minimal stimulation in vitro fertilization (mini-IVF consists of a gentle controlled ovarian stimulation that aims to produce a maximum of five to six oocytes. There is a misbelief that mini-IVF severely compromises pregnancy and live birth rates. An appraisal of the literature pertaining to studies on mini-IVF protocols was performed. The advantages of minimal stimulation protocols are reported here with a focus on the use of clomiphene citrate (CC, gonadotropin releasing hormone (GnRH ago- nist trigger for oocyte maturation, and freeze-all embryo strategy. Literature review and the author’s own center data suggest that minimal ovarian stimulation protocols with GnRH agonist trigger and freeze-all embryo strategy along with single embryo transfer produce a reasonable clinical pregnancy and live birth rates in both good and poor responders. Additionally, mini-IVF offers numerous advantages such as: i. Reduction in cost and stress with fewer office visits, needle sticks, and ultrasounds, and ii. Reduction in the incidence of ovarian hyperstimulation syndrome (OHSS. Mini-IVF is re-emerging as a solution for some of the problems associated with conventional IVF, such as OHSS, cost, and patient discomfort.

  7. Culture of bovine embryos on a polydimethylsiloxane (PDMS) microwell plate.

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    Akagi, Satoshi; Hosoe, Misa; Matsukawa, Kazutsugu; Ichikawa, Akihiko; Tanikawa, Tamio; Takahashi, Seiya

    2010-08-01

    We fabricated a polydimethylsiloxane (PDMS)-based microwell plate (PDMS-MP) containing 100 microwells with a rounded bottom and examined whether it can be used for culture of individual in vitro fertilized (IVF) embryos or parthenogenetically activated zona-free embryos in cattle. In Experiment 1, we examined the in vitro developmental ability of IVF embryos cultured individually on PDMS-MP. After IVF, 20 embryos were transferred into 100 microl drops on PDMS-MP and cultured individually in each well of PDMS-MP (PDMS group). After 7 days of culture, the embryos in the PDMS group developed to the blastocyst stage at the same rate of those in the control group cultured in a group of 20 embryos without PDMS-MP. There were no differences in total number of cells and the ratio of inner cell mass to total cells between the PDMS and control groups. In Experiment 2, we examined the in vitro developmental ability of parthenogenetically activated zona-free bovine embryos cultured individually on PDMS-MP. The zona-free embryos were cultured individually in each well of a PDMS-MP or in each well produced by pressing a darning needle onto the bottom of a culture dish (WOW group). After 7 days of culture, the blastocyst formation rate and cell number of blastocysts in the PDMS group did not differ from those of the zona-intact embryos in the control group. Also, there were no differences in the blastocyst formation rate and cell number of blastocysts between the WOW and PDMS groups. These results suggest that the culture system using PDMS-MP is useful for individual embryos or zona-free embryos in cattle.

  8. Obstetric and perinatal outcomes after either fresh or thawed frozen embryo transfer: an analysis of 112,432 singleton pregnancies recorded in the Human Fertilisation and Embryology Authority anonymized dataset.

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    Maheshwari, Abha; Raja, Edwin Amalraj; Bhattacharya, Siladitya

    2016-12-01

    To explore obstetric and perinatal outcomes in singleton pregnancies occurring as a result of fresh and thawed frozen embryo transfer using anonymized Human Fertilisation and Embryology Authority data. Retrospective cohort study. Not applicable. Singleton births after IVF/intracytoplasmic sperm injection cycles in the United Kingdom (1991-2011). A total of 112,432 cycles (95,911 fresh and 16,521 frozen) were analyzed using multivariate logistic regression to explore associations between type of embryo transferred (frozen vs. fresh) and obstetric and perinatal outcomes. Relative risks (RRs) and their 99.5% confidence intervals (CIs) were calculated using Stata 14 MP, adjusting for potential confounders. Birth weight. The adjusted RR (99.5% CI) of low birth weight [0.73 (0.66-0.80)] and very low birth weight [0.78 (0.63-0.96)] were all lower after frozen embryo transfer; however, RR of having a high birth weight baby was higher [1.64 (1.53-1.76)]. There was no difference in RR of preterm birth [0.96 (0.88-1.03)], very preterm birth [0.86 (0.70-1.05)], and congenital anomalies [0.86 (0.73-1.01)]. The findings of low birth and very low birth weight after thawed frozen embryo transfer are consistent with the literature and provide reassurance regarding the outcome of pregnancies after frozen embryo transfers. However, they highlight the possibility of high birth weight in these babies. Because these results are based on observational data, further evidence from randomized, controlled trials is needed before elective cryopreservation of all embryos is practiced in preference to the current practice of transfer of fresh embryos. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

  9. Evaluation of serum estradiol/ progestrone ratio on day of embryo transfer and it’s effect on intra-cytoplasmic sperm induction outcome

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    Masomeh Hagshafiha

    2013-07-01

    Full Text Available Background : One of the important problems in fertilization in vitro (IVF is failure of implantation. This could be the result of estrogen and progesterone effects in endometrial acceptance during ovulation stimulation. Although progesterone has a vital role in primary phase of pregnancy, but the estradiol role in luteal phase is unknown. The aim of this study is assessment of the ratio of estradiol to progesterone in embryo transfer day on Intracytoplasmic sperm injection (ICSI outcomes. Methods : This is a cohort study. The subjects were 311 infertile women referred to Urmia Kosar infertility clinic & Urmia reproductive health research center who treated with ICSI method between August-Jan 2011 . Five cc blood was drawn for determine of estradiol and progesterone in transfer day. Transfer occurred after a variable in vitro culture period ranging from 48 to 72 hours after ovulation induction. Chemiluminescent ELICA the level of mention hormones was used to determine the ratio of the progesterone level to the estradiol serum level and was compared based on treatment outcomes. Results : A total of 311 patients, 115 (37% were pregnant and happened abortion were 18 (5.8% . The mean ratio of estradiol to progesterone in transfer day in two groups of miscarriage/ non miscarriage and pregnant/ non pregnant was 32.26±23.86 , 28.17±26.5 and 28.58±2.4 , 36.09±4.39 respectively. There is no significant difference between two groups in regard of estradiol on progesterone ratio (P=0.5, P= 0.2 . Conclusion: The results of this prospective cohort study show that there is no effect of estradiol to progesterone ratio on day of embryo transfer, successful pregnancy in ICSI cycles and abortion rate following of ICSI .

  10. The effect on human sex ratio at birth by assisted reproductive technology (ART) procedures--an assessment of babies born following single embryo transfers, Australia and New Zealand, 2002-2006.

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    Dean, J H; Chapman, M G; Sullivan, E A

    2010-12-01

    To assess the effect on the human sex ratio at birth by assisted reproductive technology (ART) procedures. Retrospective population-based study. Fertility clinics in Australia and New Zealand. The study included 13,368 babies by 13,165 women who had a single embryo transfer (SET) between 2002 and 2006. Logistic regression was used to model the effect on the sex ratio at birth of ART characteristics [in vitro fertilisation (IVF) or intracytoplasmic sperm insemination (ICSI) SET, cleavage-stage or blastocyst SET, and fresh or thawed SET] and biological characteristics (woman's and partner's age and cause of infertility). Proportion of male births. The crude sex ratio at birth was 51.3%. Individual ART procedures had a significant effect on the sex ratio at birth. More males were born following IVF SET (53.0%) than ICSI SET (50.0%), and following blastocyst SET (54.1%) than cleavage-stage SET (49.9%). For a specific ART regimen, IVF blastocyst SET produced more males (56.1%) and ICSI cleavage-stage SET produced fewer males (48.7%). The change in the sex ratio at birth of SET babies is associated with the ART regimen. The mechanism of these effects remains unclear. Fertility clinics and patients should be aware of the bias in the sex ratio at birth when using ART procedures. © 2010 The Authors Journal compilation © RCOG 2010 BJOG An International Journal of Obstetrics and Gynaecology.

  11. Infertility treatment strategy involving combined freeze-all embryos and single vitrified-warmed embryo transfer during hormonal replacement cycle for in vitro fertilization of women with hypogonadotropic hypogonadism.

    Science.gov (United States)

    Kuroda, Keiji; Ezoe, Kenji; Kato, Keiichi; Yabuuchi, Akiko; Segawa, Tomoya; Kobayashi, Tamotsu; Ochiai, Asako; Katoh, Noriko; Takeda, Satoru

    2018-02-13

    Hypogonadotropic hypogonadism (HH) is a condition caused by the deficient secretion of pituitary gonadotropins, leading to diminished ovarian function. Several studies of in vitro fertilization (IVF) in women with HH revealed acceptable clinical pregnancy outcomes but high multiple pregnancy rates after multiple fresh embryo transfer (ET). The purpose of this study was to analyze the outcomes of combined freeze-all embryos and single vitrified-warmed ET in women with HH. Of 91 infertile women with HH (basal luteinizing hormone and follicle-stimulating hormone levels <2.0 mIU/mL), we excluded patients aged ≥40 years (n = 2) and women who preferred fresh ET (n = 10). Seventy-nine women underwent 117 oocyte retrieval cycles and 135 vitrified-warmed ET during hormone replacement (HR) cycles from 2008 to 2014 at the Kato Ladies Clinic and Juntendo University Hospital. In 26 single cleavage ET cycles, the rates of clinical pregnancy and live birth were 34.6% (9/26 ET) and 26.9% (7/26 ET), respectively. Regarding the outcomes after single vitrified-warmed blastocyst transfer, clinical pregnancy and live birth rates were 65.1% (71/109 ET) and 50.5% (55/109 ET), respectively. Multiple conceptions and ovarian hyperstimulation syndrome did not occur in any of the women with HH. Our results demonstrated that IVF followed by single vitrified-warmed ET in adjusted endocrine milieu during the HR cycle is an effective fertility treatment for women with HH and decreases the incidence of complications, including multiple conceptions. © 2018 Japan Society of Obstetrics and Gynecology.

  12. Growth hormone gene family (GH, GHR, GHRH and Pit-1 polymorphisms and its association with superovulation response, ovulation rate and embryo quality in Embryo Transfer Station (BET of Cipelang

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    Cece Sumantri

    2011-06-01

    Full Text Available The decrease in fertility is considered to be the main cause of reproductive loss in dairy cattle and beef industry. Many candidate genes that play an important role in fertility and embryonic development. The purpose of this study was to detect genetic variations of the growth hormone gene family (GH|MspI, GH|AluI, GHR| AluI, GHRH|HaeIII and Pit-1|HinfI and its association with superovulation response, ovulation, fertilization and transferable embryos rate. A total of 45 blood samples taken from cows that have been superovulated Angus, Brahman, HF, Limousin and Simmental. DNA was extracted with phenol-chloroform protocol followed by polymerase chain reaction technique (PCR using specific primers for GH, GHR, GHRH and Pit-1 gene. PCR product was cut with restriction enzyme MspI, AluI, HaeIII and HinfI and electrophoresed on agarose gel and stained with ethidium bromide (EtBr. Superovulation is done by injecting a totally of 20 ml FSH for 4 days. Injecting the prostaglandin hormone (PGF2α was performed on the eleventh day of CIDR implantation. Artificial insemination (AI performed two or three days after the injection of PGF2α and Flushing was done on the seventh day after the AI. The results showed that the Angus, Limousin, Brahman and Simental GH loci diversity of GH|MspI, GH|AluI, GHR|AluI, GHRH|HaeIII and Pit-1|HinfI was not associated with superovulation response, ovulation, fertilization and transferable embryo rate. In HF dairy cattle, genotype on Pit-1|HinfI AA has higher percentage of superovulation response (P < 0.05 when compared to AB genotype, but did not differ to BB genotype. Dairy cattle HF AA genotype also had higher ovulation rate (P < 0.05 when compared to AB and BB genotypes, but AB and BB have the same ovulation rate.

  13. Stem Cell Differentiation Stage Factors from Zebrafish Embryo: A Novel Strategy to Modulate the Fate of Normal and Pathological Human (Stem) Cells.

    Science.gov (United States)

    Biava, Pier M; Canaider, Silvia; Facchin, Federica; Bianconi, Eva; Ljungberg, Liza; Rotilio, Domenico; Burigana, Fabio; Ventura, Carlo

    2015-01-01

    In spite of the growing body of evidence on the biology of the Zebrafish embryo and stem cells, including the use of Stem Cell Differentiation Stage Factors (SCDSFs) taken from Zebrafish embryo to impact cancer cell dynamics, comparatively little is known about the possibility to use these factors to modulate the homeostasis of normal human stem cells or to modulate the behavior of cells involved in different pathological conditions. In the present review we recall in a synthetic way the most important researches about the use of SCDSFs in reprogramming cancer cells and in modulating the high speed of multiplication of keratinocytes which is characteristic of some pathological diseases like psoriasis. Moreover we add here the results about the capability of SCDSFs in modulating the homeostasis of human adiposederived stem cells (hASCs) isolated from a fat tissue obtained with a novel-non enzymatic method and device. In addition we report the data not yet published about a first protein analysis of the SCDSFs and about their role in a pathological condition like neurodegeneration.

  14. In Vitro Embryo as a Person

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    محمد حسن صادقی مقدم

    2016-03-01

    Full Text Available Based on moral principles and the natural laws, an in vitro embryo is considered as a natural embryo. The difference in the course of natural growing of an in vitro embryo and natural embryo, does not result in a difference of the governing rules, since both are embryos. It is argued that an in vitro embryo has the same rights as the natural embryo. Considering this claim, despite the position of some scholars who believe that such an embryo is subject to ownership, this article aims to critique this idea and argue that an in vitro embryo has the same personhood as a natural embryo from the moment of conception. Therefore, the embryo cannot be transferred as a gift or otherwise traded in the form of a stipulation in a contract.

  15. In vitro fertilization outcome in frozen versus fresh embryo transfer in women with elevated progesterone level on the day of HCG injection: An RCT

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    Marzieh Aghahosseini

    2017-12-01

    Full Text Available Background: The effect of elevated progesterone level on human chorionic gonadotropin (HCG day in in vitro fertilization cycles is controversial. Some suppose that rise in progesterone level seems to have a negative impact on implantation and pregnancy by desynchronizing the endometrium, while others disagree. Objective: To evaluate the superiority of the frozen cycle over fresh cycle on live birth in patients with elevated progesterone level on HCG day. Materials and Methods: In this double-blind, randomized clinical trial, 72 women undergoing assisted reproductive technology with elevated progesterone level (≥1.8 ng/dl on HCG day were included. The participants were grouped by fresh versus frozen embryo transfer, randomly. Finally, the clinical pregnancy and live birth rate were compared. Results: The implantation rate was 21.51%. The clinical pregnancy rate was 47.22% in fresh embryo transfer group (17/36 and 41.66% in frozen group (15/36 (p=0. 40. The live birth rate was not significantly difference between two groups (p=0.56. Conclusion: None of the fresh and frozen cycles are superior to the other and we recommend individualizing the decision for each patient. The frozen cycle may impose more emotional stress on patients

  16. Effect of music therapy on the anxiety levels and pregnancy rate of women undergoing in vitro fertilization-embryo transfer: A randomized controlled trial.

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    Aba, Yilda Arzu; Avci, Dilek; Guzel, Yilmaz; Ozcelik, Semanur Kumral; Gurtekin, Basak

    2017-08-01

    The aim of this study was to determine the effect of music therapy on the anxiety levels and pregnancy rates of women who underwent in vitro fertilization-embryo transfer. This prospective randomized controlled trial was conducted with 186 infertile women who presented to the In Vitro Fertilization Unit at the American Hospital in Turkey between April 2015 and April 2016. The infertile women who met the inclusion criteria were assigned to the music therapy group or the standard therapy group through block randomization. The study data were collected using the Personal Information Form, and State-Trait Anxiety Inventory. Early treatment success was determined by serum beta human chorionic gonadotrophin levels seven or ten days after the luteal day zero. For the analysis, descriptive statistics, chi-square test, Fisher's exact test, independent sample t-test were used. After the embryo transfer, the mean state anxiety scores decreased in both groups, and the mean trait anxiety score decreased in the music therapy group; however, the difference was not statistically significant (p>0.05). Clinical pregnancy rates did not differ between the music (48.3%) and standard (46.4%) therapy groups. After the two sessions of music therapy, state and trait anxiety levels decreased and pregnancy rates increased, but the difference was not significant. Therefore, larger sample sizes and more sessions are needed to evaluate whether music therapy has an effect on clinical outcomes. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Effects of Insulin-like Growth Factor-1 on Development of Somatic Cell Cloned Bovine Embryos.

    Science.gov (United States)

    Qu, Pengxiang; Li, Yanyan; Deng, Tengfei; Jia, Dan; Qing, Suzhu; Su, Jianmin; Zhang, Yong; Wang, Yongsheng

    2016-06-01

    The aim of this study was to assess the effect of insulin-like growth factor-1 (IGF-1) on the developmental competence of somatic cell nuclear transfer (SCNT) bovine embryos. First, the expression levels of IGF-1 receptor (IGF-1R) and IGF-1 in the oocytes and embryos of different developmental stages were examined. Then the effects of exogenous IGF-1 on the development of SCNT embryos were evaluated both in vitro and in vivo. The results showed that IGF-1 was not expressed in both IVF and SCNT embryos, whereas IGF-1R could be detected throughout the preimplantation stages in both protein and mRNA levels. Also, exogenous IGF-1 had no obvious impact on the developmental competence of IVF embryos. However, it could improve the developmental competence of SCNT embryos in terms of blastocyst developmental rate (31.3% vs. 43.2%, p embryo transfer, there was an upward tendency in both examined terms in the IGF-1-supplemented group when compared with the control group. In conclusion, the present study showed that supplementing exogenous IGF-1 to the culture medium has an obvious positive effect on the development competence of SCNT embryos.

  18. Blastocyst Morphology Holds Clues Concerning The Chromosomal Status of The Embryo

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    Rita de Cassia Savio Figueira

    2015-07-01

    Full Text Available Background: Embryo morphology has been proposed as an alternative marker of chromosomal status. The objective of this retrospective cohort study was to investigate the association between the chromosomal status on day 3 of embryo development and blastocyst morphology. Materials and Methods: A total of 596 embryos obtained from 106 cycles of intracytoplasmic sperm injection (ICSI followed by preimplantation genetic aneuploidy screening (PGS were included in this retrospective study. We evaluated the relationship between blastocyst morphological features and embryonic chromosomal alteration. Results: Of the 564 embryos with fluorescent in situ hybridization (FISH results, 200 reached the blastocyst stage on day 5 of development. There was a significantly higher proportion of euploid embryos in those that achieved the blastocyst stage (59.0% compared to embryos that did not develop to blastocysts (41.2% on day 5 (P<0.001. Regarding blastocyst morphology, we observed that all embryos that had an abnormal inner cell mass (ICM were aneuploid. Embryos with morphologically normal ICM had a significantly higher euploidy rate (62.1%, P<0.001. As regards to the trophectoderm (TE morphology, an increased rate of euploidy was observed in embryos that had normal TE (65.8% compared to embryos with abnormal TE (37.5%, P<0.001. Finally, we observed a two-fold increase in the euploidy rate in high-quality blastocysts with both high-quality ICM and TE (70.4% compared to that found in low-quality blastocysts (31.0%, P<0.001. Conclusion: Chromosomal abnormalities do not impair embryo development as aneuploidy is frequently observed in embryos that reach the blastocyst stage. A high-quality blastocyst does not represent euploidy of chromosomes 13, 14, 15, 16, 18, 21, 22, X and Y. However, aneuploidy is associated with abnormalities in the ICM morphology. Further studies are necessary to confirm whether or not the transfer of blastocysts with low-quality ICM should be

  19. The effect of laser-assisted hatching on pregnancy outcomes of cryopreserved-thawed embryo transfer: a meta-analysis of randomized controlled trials.

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    Zeng, MeiFang; Su, SuQin; Li, LiuMing

    2018-04-01

    It is well known that laser-assisted hatching (LAH) is the most popular and ideal embryo hatching technology, but the relevance to pregnancy outcomes of cryopreserved-thawed embryo transfer (ET) is controversial. The purpose of this meta-analysis was to evaluate the effects of LAH on pregnancy outcomes of cryopreserved-thawed ET. We searched for relevant studies published in the PubMed, EMBASE, and Cochrane Central databases up to March 2017. This meta-analysis was primarily used to evaluate the effect of laser-assisted hatching on assisted reproductive outcomes: clinical pregnancy, embryo implantation, multiple pregnancy, miscarriage, and live birth. Using the Mantel-Haenszel fixed effects model and random effects model, we determined the summary odds ratios (OR) with 95% confidence intervals (CIs). There were 12 randomized controlled trials (more than 2574 participants) included in our analysis. The rates of clinical pregnancy (OR = 1.65, 95% CI = 1.24-2.19, I 2  = 49), implantation (OR = 1.59, 95% CI = 1.06-2.38, I 2  = 82%), multiple pregnancy (OR = 2.30, 95% CI = 1.30-4.07, I 2  = 33%), miscarriage (OR = 0.86, 95% CI = 0.50-1.48, I 2  = 0%), and live birth (OR = 1.09, 95% CI = 0.77-1.54, I 2  = 0%) revealed comparable results for both groups. In summary, this meta-analysis demonstrates that LAH is related to a higher clinical pregnancy rate, embryo implantation rate, and multiple pregnancy rate in women with cryopreserved-thawed embryos. However, LAH is unlikely to increase live birth rates and miscarriage rates. Due to the small sample evaluated in the pool of included studies, large-scale, prospective, randomized, controlled trials are required to determine if these small effects are clinically relevant.

  20. Syntactic transfer in the initial stages of adult third language and fourth language acquisition

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    Mahbube Tavakol

    2016-01-01

    Full Text Available This paper elucidates the articulated proposals for the initial stages of adult third language (L3 syntactic transfer, addressing their application for L3 and the subsequent fourth language (L4 acquisition. The study was set to demonstrate empirical evidence in line with or against the tenets of the models and to indicate if and how syntactic transfer might obtain differently depending on the language being acquired– L3 vs. L4. The models to be tested were Full Transfer/Full Access (FT/FA, L2 Status Factor Hypothesis (LSFH, Cumulative Enhancement Model (CEM and Typological Primacy Model (TPM. Following a principles and parameters framework, six parameters were selected to generate several language pairings and an adult female’s L3 Italian and L4 German’s early spontaneous productions of the selected features were audio-recorded. The accuracy levels with which the features were produced in tandem with the results of error analyses violated the positions of FT/FA as considered for L3/s acquisition and CEM and consistently identified Typological proximity and L2 status as affecting syntactic transfer during the early stages multilingual acquisition.

  1. The Well-of-the-Well system: an efficient approach to improve embryo development.

    Science.gov (United States)

    Vajta, Gábor; Korösi, Tamás; Du, Yutao; Nakata, Kumiko; Ieda, Shoko; Kuwayama, Masashige; Nagy, Zsolt Peter

    2008-07-01

    Transfer of human embryos at the blastocyst stage may offer considerable benefits including an increased implantation rate and a decreased risk of multiple pregnancies; however, blastocyst culture requires an efficient and reliable in-vitro embryo culture system. In this study, the effect of the Well-of-the-Well (WOW) system consisting of microwells formed on the bottom of the culture dish was tested in three mammalian species, including humans. The WOW system resulted in significant improvement when comparing the drops for culture of in-vitro-matured and parthenogenetically activated porcine oocytes, and in-vivo-derived mouse zygotes. In human embryos, using a sibling oocyte design, embryos cultured in WOW developed to the blastocyst stage in a significantly higher proportion than did embryos cultured traditionally (55% in WOW and 37% in conventional culture; P WOW system or in microdrops. Transferable quality blastocyst development (48.9% of cultured zygotes) was observed in the WOW system. Ninety-four blastocysts transferred to 45 patients resulted in clinical pregnancy rates of 48.9%, including nine twin pregnancies, seven single pregnancies, five miscarriages and one ectopic pregnancy. The results indicate that the WOW system provides a promising alternative for microdrop culture of mammalian embryos, including human embryos.

  2. [An ongoing twin pregnancy after embryo time laps monitoring in a patient with a history of IVF failures--case report and literature review].

    Science.gov (United States)

    Radwan, Paweł; Krasiński, Rafał; Radwan, Michał; Połać, Ireneusz

    2014-04-01

    A standard assessment of embryo morphology at given time points does not always allow to transfer the embryo with the highest implantation potential. The effect of transfer of an improper embryo results in a lack of pregnancy or a miscarriage and, as a consequence, exposes the patient to unnecessary emotional stress and necessity to perform yet another transfer of frozen embryos. We present a case of a patient with earlier IVF failures. The use of time-lapse technique in this case helped to choose two good embryos. The transfer resulted in ongoing twin pregnancy. A 35-year-old woman with history of IVF-ET treatment failure was deemed eligible for an ICSI procedure because of the male factor. Ovarian stimulation was performed according to the agonist long protocol. Eight MII oocytes were fertilized and seven embryos were obtained. Continuous embryo monitoring was performed with the use of Primo Vision system. Forty-four hours after fertilization only 2 correctly developing embryos were identified. They were transferred on day 3. The development of the remaining 5 embryos was arrested. These embryos did not achieve the blastocyst stage on day 5-6 after fertilization. Forty days after embryo transfer a twin pregnancy, confirmed with fetal heart rate of both fetuses, was revealed on ultrasound examination. Currently the patient is at 27 weeks of ongoing twin gestation. The system of continuous embryo monitoring introduces new criteria for the examination of embryo development. These new parameters can be useful in clinical practice. However prospective randomized studies are necessary to provide data confirming the usefulness of time-lapse technique in IVF treatment.

  3. Soluble CD146, an innovative and non-invasive biomarker of embryo selection for in vitro fertilization.

    Directory of Open Access Journals (Sweden)

    Sylvie Bouvier

    Full Text Available Although progress was made in in vitro fertilization (IVF techniques, the majority of embryos transferred fail to implant. Morphology embryo scoring is the standard procedure for most of IVF centres for choosing the best embryo, but remains limited since even the embryos classified as "top quality" may not implant. As it has been shown that i CD146 is involved in embryo implantation and ii membrane form is shed to generate soluble CD146 (sCD146, we propose that sCD146 in embryo supernatants may constitute a new biomarker of embryo selection. Immunocytochemical staining showed expression of CD146 in early embryo stages and sCD146 was detected by ELISA and Western-blot in embryo supernatants from D2. We retrospectively studied 126 couples who underwent IVF attempt. The embryo culture medium from each transferred embryo (n = 222 was collected for measurement of sCD146 by ELISA. Significantly higher sCD146 concentrations were present in embryo supernatants that did not implant (n = 185 as compared to those that successfully implanted (n = 37 (1310 +/- 1152 pg.mL-1 vs. 845+/- 1173 pg.mL-1, p = 0.024. Sensitivity analysis performed on single embryo transfers (n = 71 confirmed this association (p = 0.0054. The computed ROC curve established that the optimal sCD146 concentration for embryo implantation is under 1164 pg.mL-1 (sensitivity: 76%, specificity: 48%, PPV: 25% and NPV: 92%. Over this sCD146 threshold, the implantation rate was significantly lower (9% with sCD146 levels >1164 pg.ml-1 vs. 22% with sCD146 levels ≤ 1164 pg.mL-1, p = 0.01. Among the embryos preselected by morphologic scoring, sCD146 determination could allow a better selection of the embryo(s, thus improving the success of elective single embryo transfer. This study establishes the proof of concept for the use of sCD146 as a biomarker for IVF by excluding the embryo with the highest sCD146 level. A multicentre prospective study will now be necessary to further establish its use in

  4. Single-stage reconstruction of flexor tendons with vascularized tendon transfers.

    Science.gov (United States)

    Cavadas, P C; Pérez-García, A; Thione, A; Lorca-García, C

    2015-03-01

    The reconstruction of finger flexor tendons with vascularized flexor digitorum superficialis (FDS) tendon grafts (flaps) based on the ulnar vessels as a single stage is not a popular technique. We reviewed 40 flexor tendon reconstructions (four flexor pollicis longus and 36 finger flexors) with vascularized FDS tendon grafts in 38 consecutive patients. The donor tendons were transferred based on the ulnar vessels as a single-stage procedure (37 pedicled flaps, three free flaps). Four patients required composite tendon and skin island transfer. Minimum follow-up was 12 months, and functional results were evaluated using a total active range of motion score. Multiple linear regression analysis was performed to evaluate the factors that could be associated with the postoperative total active range of motion. The average postoperative total active range of motion (excluding the thumbs) was 178.05° (SD 50°). The total active range of motion was significantly lower for patients who were reconstructed with free flaps and for those who required composite tendon and skin island flap. Age, right or left hand, donor/motor tendon and pulley reconstruction had no linear effect on total active range of motion. Overall results were comparable with a published series on staged tendon grafting but with a lower complication rate. Vascularized pedicled tendon grafts/flaps are useful in the reconstruction of defects of finger flexor tendons in a single stage, although its role in the reconstructive armamentarium remains to be clearly established. © The Author(s) 2014.

  5. Antioxidants improve mouse preimplantation embryo development and viability.

    Science.gov (United States)

    Truong, Thi T; Soh, Yu May; Gardner, David K

    2016-07-01

    What is the effect of three antioxidants (acetyl-L-carnitine, N-acetyl-L-cysteine and α-lipoic acid), when used individually and in combination, on mouse embryo development in culture, and subsequent fetal development post-transfer? A combination of antioxidants resulted in significant increases in blastocyst cell number, maintained intracellular glutathione (GSH) levels, supported earlier cleavage times from 5-cell stage to expanded blastocyst, and improved fetal developmental irrespective of incubator oxygen concentration. Acetyl-L-carnitine, N-acetyl-L-cysteine and α-lipoic acid have been shown to have beneficial effects individually in several tissues, and most recently on developing embryos, in the presence of oxidative stress. Morphokinetics of mouse embryos were quantitated using time-lapse imaging. GSH levels in pronucleate oocytes were measured. Blastocysts underwent differential nuclear staining for inner cell mass and trophectoderm cells or were transferred to recipient females to assess implantation and fetal development. Pronucleate oocytes from F1 mice were cultured in 5 or 20% oxygen either individually or in groups of 10, in media G1/G2, in the presence or absence of 10 µM acetyl-L-carnitine /10 µM N-acetyl-L-cysteine /5 µM α-lipoic acid, either individually or in combination. Controls were embryos cultured without antioxidants. Intracellular levels of reduced glutathione were quantitated in pronucleate oocytes. Embryo development and viability were analysed through time-lapse microscopy and embryo transfers. Antioxidants significantly increased mouse blastocyst cell numbers compared with control when used individually (Pantioxidants resulted in faster development rates to 5-cell cleavage stage, which continued until the expanded blastocyst stage when cultured in 20% oxygen. The beneficial effects of combining the antioxidants were greater for embryos cultured individually as opposed to in groups of 10 and for those embryos cultured in 20

  6. From callus to embryo: a proteomic view on the development and maturation of somatic embryos in Cyclamen persicum.

    Science.gov (United States)

    Rode, Christina; Lindhorst, Kathrin; Braun, Hans-Peter; Winkelmann, Traud

    2012-05-01

    In this study, the proteome structures following the pathway in somatic embryogenesis of Cyclamen persicum were analysed via high-resolution 2D-SDS-PAGE with two objectives: (1) to identify the significant physiological processes during somatic embryogenesis in Cyclamen and (2) to improve the maturation of somatic embryos. Therefore, the effects of maturation-promoting plant growth regulator abscisic acid (ABA) and high sucrose levels on torpedo-shaped embryos were investigated. In total, 108 proteins of differential abundance were identified using a combination of tandem mass spectrometry and a digital proteome reference map. In callus, enzymes related to energy supply were especially distinct, most likely due to energy demand caused by fast growth and cell division. The switch from callus to globular embryo as well as from globular to torpedo-shaped embryo was associated with controlled proteolysis via the ubiquitin-26S proteasome pathway. Storage compound accumulation was first detected 21 days after transfer to plant growth regulator (PGR)-free medium in early torpedo-shaped embryos. Increase in abundance of auxin-amidohydrolase during embryogenesis suggests a possible increase in auxin release in the late embryo stages of Cyclamen. A development-specific isoelectric point switch of catalases has been reported for the first time for somatic embryogenesis. Several proteins were identified to represent markers for the different developmental stages analysed. High sucrose levels and ABA treatment promoted the accumulation of storage compounds in torpedo-shaped embryos. Additionally, proteins of the primary metabolic pathways were decreased in the proteomes of ABA-treated embryos. Thus, ABA and high sucrose concentration in the culture medium improved maturation and consequently the quality of somatic embryos in C. persicum.

  7. A Randomized Trial to Evaluate the Effect of Local Endometrial Injury on the Clinical Pregnancy Rate of Frozen Embryo Transfer Cycles in Patients With Repeated Implantation Failure

    Directory of Open Access Journals (Sweden)

    Ensieh Shahrokh-Tehraninejad

    2016-12-01

    Full Text Available Objective: Repeated implantation failure (RIF is a condition in which the embryos implantation decreases in the endometrium. So, our aim was to evaluate the effect of local endometrial injury on embryo transfer results.Materials and methods: In this simple randomized clinical trial (RCT, a total of 120 patients were selected. The participants were less than 40 years old, and they are in their minimum two cycles of vitro fertilization (IVF. Patients were divided randomly into two groups of LEI (Local endometrial injury and a control group (n = 60 in each group. The first group had four small endometrial injuries from anterior, posterior, and lateral uterus walls which were obtained from people who were in 21th day of their previous IVF cycle. The second group was the patients who have not received any intervention.Results: The experimental and control patients were matched in the following factors. Regarding the clinical pregnancy rate, there was no significant difference noted between the experimental and the control group.Conclusion: Local endometrial injury in a preceding cycle does not increase the clinical pregnancy rate in the subsequent FET cycle of patients with repeated implantation failure.

  8. A comparison of different database technologies for the CMS AsyncStageOut transfer database

    Science.gov (United States)

    Ciangottini, D.; Balcas, J.; Mascheroni, M.; Rupeika, E. A.; Vaandering, E.; Riahi, H.; Silva, J. M. D.; Hernandez, J. M.; Belforte, S.; Ivanov, T. T.

    2017-10-01

    AsyncStageOut (ASO) is the component of the CMS distributed data analysis system (CRAB) that manages users transfers in a centrally controlled way using the File Transfer System (FTS3) at CERN. It addresses a major weakness of the previous, decentralized model, namely that the transfer of the user’s output data to a single remote site was part of the job execution, resulting in inefficient use of job slots and an unacceptable failure rate. Currently ASO manages up to 600k files of various sizes per day from more than 500 users per month, spread over more than 100 sites. ASO uses a NoSQL database (CouchDB) as internal bookkeeping and as way to communicate with other CRAB components. Since ASO/CRAB were put in production in 2014, the number of transfers constantly increased up to a point where the pressure to the central CouchDB instance became critical, creating new challenges for the system scalability, performance, and monitoring. This forced a re-engineering of the ASO application to increase its scalability and lowering its operational effort. In this contribution we present a comparison of the performance of the current NoSQL implementation and a new SQL implementation, and how their different strengths and features influenced the design choices and operational experience. We also discuss other architectural changes introduced in the system to handle the increasing load and latency in delivering output to the user.

  9. A Comparison of Different Database Technologies for the CMS AsyncStageOut Transfer Database

    Energy Technology Data Exchange (ETDEWEB)

    Ciangottini, D. [INFN, Perugia; Balcas, J. [Caltech; Mascheroni, M. [Fermilab; Rupeika, E. A. [Vilnius U.; Vaandering, E. [Fermilab; Riahi, H. [CERN; Silva, J. M.D. [Sao Paulo, IFT; Hernandez, J. M. [Madrid, CIEMAT; Belforte, S. [INFN, Trieste; Ivanov, T. T. [Sofiya U.

    2017-11-22

    AsyncStageOut (ASO) is the component of the CMS distributed data analysis system (CRAB) that manages users transfers in a centrally controlled way using the File Transfer System (FTS3) at CERN. It addresses a major weakness of the previous, decentralized model, namely that the transfer of the user’s output data to a single remote site was part of the job execution, resulting in inefficient use of job slots and an unacceptable failure rate. Currently ASO manages up to 600k files of various sizes per day from more than 500 users per month, spread over more than 100 sites. ASO uses a NoSQL database (CouchDB) as internal bookkeeping and as way to communicate with other CRAB components. Since ASO/CRAB were put in production in 2014, the number of transfers constantly increased up to a point where the pressure to the central CouchDB instance became critical, creating new challenges for the system scalability, performance, and monitoring. This forced a re-engineering of the ASO application to increase its scalability and lowering its operational effort. In this contribution we present a comparison of the performance of the current NoSQL implementation and a new SQL implementation, and how their different strengths and features influenced the design choices and operational experience. We also discuss other architectural changes introduced in the system to handle the increasing load and latency in delivering output to the user.

  10. Maternal KIR haplotype influences live birth rate after double embryo transfer in IVF cycles in patients with recurrent miscarriages and implantation failure.

    Science.gov (United States)

    Alecsandru, D; Garrido, N; Vicario, J L; Barrio, A; Aparicio, P; Requena, A; García-Velasco, J A

    2014-12-01

    In patients with recurrent miscarriages (RM) or recurrent implantation failure (RIF), does the maternal killer immunoglobulin-like receptor (KIR) haplotype have an impact on live birth rates per cycle after embryo transfer with the patient's own or donated oocytes? After double embryo transfer (DET) in patients with the maternal KIR AA haplotype, a significantly increased early miscarriage rate was observed when the patient's own oocytes were used, and a significantly decreased live birth rate per cycle after embryo transfer was observed when donated oocytes were used. Interactions between fetal HLA-C and maternal KIR influence placentation during human pregnancy. There is an increased risk of RM, pre-eclampsia or fetal growth restriction in mothers with the KIR AA haplotype when the fetus has more HLA-C2 genes than the mother. Between 2010 and 2014, we performed a retrospective study that included 291 women, with RM or RIF, who had a total of 1304 assisted reproductive cycles. Pregnancy, miscarriage and live birth rates per cycle after single or DET, categorized by the origin of the oocytes and the presence of maternal KIR haplotypes, were studied. KIR haplotype regions were defined by the presence of the following KIR genes: Cen-A/2DL3; Tel-A/3DL1 and 2DS4; Cen-B/2DL2 and 2DS2; as well as Tel-B/2DS1 and 3DS1. Higher rates of early miscarriage per cycle after DET with the patient's own oocytes in mothers with the KIR AA haplotype (22.8%) followed by those with the KIR AB haplotype (16.7%) compared with mothers with the KIR BB haplotype (11.1%) were observed (P = 0.03). Significantly decreased live birth rates per cycle were observed after DET of donated oocytes in mothers with the KIR AA haplotype (7.5%) compared with those with the KIR AB (26.4%) and KIR BB (21.5%) haplotypes (P = 0.006). No statistically significant differences were observed for pregnancy, miscarriage and live birth rates per cycle among those with maternal KIR AA, AB and BB haplotypes after

  11. Noninvasive Digital Detection of Fetal DNA in Plasma of 4-Week-Pregnant Women following In Vitro Fertilization and Embryo Transfer.

    Directory of Open Access Journals (Sweden)

    Bedri Karakas

    Full Text Available The discovery of cell-free fetal DNA (cfDNA circulating in the maternal blood has provided new opportunities for noninvasive prenatal diagnosis (NIPD. However, the extremely low levels of cfDNA within a high background of the maternal DNA in maternal circulation necessitate highly sensitive molecular techniques for its reliable use in NIPD. In this proof of principle study, we evaluated the earliest possible detection of cfDNA in the maternal plasma by a bead-based emulsion PCR technology known as BEAMing (beads, emulsion, amplification, magnetics. Blood samples were collected from in vitro fertilization (IVF patients at 2 to 6 weeks following embryo transfer (i.e., 4 to 8 week pregnancies and plasma DNA was extracted. The genomic regions of both X and Y chromosome-specific sequences (AMELX and AMELY were concurrently amplified in two sequential PCRs; first by conventional PCR then by BEAMing. The positive beads either for AMELX or AMELY gene sequences were counted by a flow cytometer. Our results showed that the pregnancies yielding boys had significantly higher plasma AMELY gene fractions (0.512 ± 0.221 than the ones yielding girls (0.028 ± 0.003 or non-pregnant women (0.020 ± 0.005, P= 0.0059. Here, we clearly demonstrated that the BEAMing technique is capable of reliably detecting cfDNA in the blood circulation of 4-week-pregnant women, which is only two weeks after the embryo transfer. BEAMing technique can also be used to early detect fetal DNA alterations in other pregnancy-associated disorders.

  12. Contrasting Storage Protein Synthesis and Messenger RNA Accumulation during Development of Zygotic and Somatic Embryos of Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Krochko, J E; Pramanik, S K; Bewley, J D

    1992-05-01

    During development on hormone-free media, somatic embryos pass through distinct morphological stages that superficially resemble those of zygotic embryo development (globular, heart, torpedo, cotyledonary stages). Despite these similarities, they differ from zygotic embryos in the extent of cotyledonary development and the patterns of synthesis and quantitative expression of seed-specific storage proteins (7S, 11S, and 2S proteins). Alfin (7S) is the first storage protein synthesized in developing zygotic embryos (stage IV). The 11S (medicagin) and 2S (Low Molecular Weight, LMW) storage proteins are not detectable until the following stage of development (stage V), although all three are present before the completion of embryo enlargement. Likewise, the 7S storage protein is the first to be synthesized in developing somatic embryos (day 5). Medicagin is evident by day 7 and the LMW protein by day 10. In contrast to zygotic embryos, alfin remains the predominant storage protein in somatic embryos throughout development. Not only are the relative amounts of medicagin and the LMW protein reduced in somatic embryos but the LMW protein is accumulated much later than the other proteins. Quantification of the storage protein mRNAs (7S, 11S, and 2S) by northern blot analysis confirms that there are substantial differences in the patterns of message accumulation in zygotic and somatic embryos of alfalfa (Medicago sativa). In zygotic embryos, the 7S, 11S, and 2S storage protein mRNAs are abundant during maturation and, in particular, during the stages of maximum protein synthesis (alfin, stages VI and VII; medicagin, stage VII; LMW, stage VII). In somatic embryos, the predominance of the 7S storage protein is correlated with increased accumulation of its mRNA, whereas the limited synthesis of the 11S storage protein is associated with much lower steady-state levels of its message. The mRNA for the LMW protein is present already by 3 days after transfer to hormone-free media

  13. PXD101 significantly improves nuclear reprogramming and the in vitro developmental competence of porcine SCNT embryos

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Jun-Xue; Kang, Jin-Dan; Li, Suo; Jin, Long; Zhu, Hai-Ying; Guo, Qing; Gao, Qing-Shan; Yan, Chang-Guo; Yin, Xi-Jun, E-mail: yinxj33@msn.com

    2015-01-02

    Highlights: • First explored that the effects of PXD101 on the development of SCNT embryos in vitro. • 0.5 μM PXD101 treated for 24 h improved the development of porcine SCNT embryos. • Level of AcH3K9 was significantly higher than control group at early stages. - Abstract: In this study, we investigated the effects of the histone deacetylase inhibitor PXD101 (belinostat) on the preimplantation development of porcine somatic cell nuclear transfer (SCNT) embryos and their expression of the epigenetic markers histone H3 acetylated at lysine 9 (AcH3K9). We compared the in vitro developmental competence of SCNT embryos treated with various concentrations of PXD101 for 24 h. Treatment with 0.5 μM PXD101 significantly increased the proportion of SCNT embryos that reached the blastocyst stage, in comparison to the control group (23.3% vs. 11.5%, P < 0.05). We tested the in vitro developmental competence of SCNT embryos treated with 0.5 μM PXD101 for various amounts of times following activation. Treatment for 24 h significantly improved the development of porcine SCNT embryos, with a significantly higher proportion of embryos reaching the blastocyst stage in comparison to the control group (25.7% vs. 10.6%, P < 0.05). PXD101-treated SCNT embryos were transferred into two surrogate sows, one of whom became pregnant and four fetuses developed. PXD101 treatment significantly increased the fluorescence intensity of immunostaining for AcH3K9 in embryos at the pseudo-pronuclear and 2-cell stages. At these stages, the fluorescence intensities of immunostaining for AcH3K9 were significantly higher in PXD101-treated embryos than in control untreated embryos. In conclusion, this study demonstrates that PXD101 can significantly improve the in vitro and in vivo developmental competence of porcine SCNT embryos and can enhance their nuclear reprogramming.

  14. Prevention of multiple pregnancies in couples with unexplained or mild male subfertility : randomised controlled trial of in vitro fertilisation with single embryo transfer or in vitro fertilisation in modified natural cycle compared with intrauterine insemination with controlled ovarian hyperstimulation

    NARCIS (Netherlands)

    Bensdorp, A. J.; Tjon-Kon-Fat, R. I.; Bossuyt, P. M. M.; Koks, C. A. M.; Oosterhuis, G. J. E.; Hoek, A.; Hompes, P. G. A.; Broekmans, F. J. M.; Verhoeve, H. R.; de Bruin, J. P.; van Golde, R.; Repping, S.; Cohlen, B. J.; Lambers, M. D. A.; van Bommel, P. F.; Slappendel, E.; Perquin, D.; Smeenk, J. M.; Pelinck, M. J.; Gianotten, J.; Hoozemans, D. A.; Maas, J. W. M.; Eijkemans, M. J. C.; van der Veen, F.; Mol, B. W. J.; van Wely, M.

    2015-01-01

    Objectives To compare the effectiveness of in vitro fertilisation with single embryo transfer or in vitro fertilisation in a modified natural cycle with that of intrauterine insemination with controlled ovarian hyperstimulation in terms of a healthy child. Design Multicentre, open label, three arm,

  15. The Periconceptional Environment and Cardiovascular Disease: Does In Vitro Embryo Culture and Transfer Influence Cardiovascular Development and Health?

    Directory of Open Access Journals (Sweden)

    Monalisa Padhee

    2015-02-01

    Full Text Available Assisted Reproductive Technologies (ARTs have revolutionised reproductive medicine; however, reports assessing the effects of ARTs have raised concerns about the immediate and long-term health outcomes of the children conceived through ARTs. ARTs include manipulations during the periconceptional period, which coincides with an environmentally sensitive period of gamete/embryo development and as such may alter cardiovascular development and health of the offspring in postnatal life. In order to identify the association between ARTs and cardiovascular health outcomes, it is important to understand the events that occur during the periconceptional period and how they are affected by procedures involved in ARTs. This review will highlight the emerging evidence implicating adverse cardiovascular outcomes before and after birth in offspring conceived through ARTs in both human and animal studies. In addition, it will identify the potential underlying causes and molecular mechanisms responsible for the congenital and adult cardiovascular dysfunctions in offspring whom were conceived through ARTs.

  16. In vitro regeneration and morphogenesis of somatic embryos of cassava

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    Ádila Melo Vidal

    Full Text Available In this study, the somatic embryogenesis and regeneration of Cigana Preta plants from shoot apices and immature leaves taken from plants cultivated in vitro were examined. To embryo induction the explants were cultivated in Murashige and Skoog (MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D or 4-amino-3,5,6-trichloropicolinic acid (Picloram at concentrations of 8.0 and 12 mg L-1. To development of embryos two media cultures with different concentration of BAP (D1 or D2 were evaluated. Embryos in the cotyledonary stage were incubated in germination medium containing MS salts and vitamins, 2.0 µM copper sulfate, 2.4 g L-1 of Phytagel® and 1.77 µM BAP. The highest frequency of calluses and the greatest number of embryos per explant were obtained using the auxin picloram at a concentration of 8.0 mg L-1. The plants regenerated in the picloram treatment exhibited normal development and were transferred to a multiplication medium after a minimum of four weeks. The histological sections of the malformed embryos from foliar explants cultured in the presence of 2,4-D demonstrated that the origin of the cotyledonary structures was independent of the formation of the shoot apical meristem, which was not formed in the embryos, and the majority of the embryos were classified as cornet-shaped. This study demonstrates that in cassava, the use of different auxins provides different conditions for the formation of somatic embryos, and the low rate of conversion into plants results from abnormalities in the embryos.

  17. The use of morphokinetic parameters to select all embryos with full capacity to implant

    OpenAIRE

    Chamayou, Sandrine; Patrizio, Pasquale; Storaci, Giorgia; Tomaselli, Venera; Alecci, Carmelita; Ragolia, Carmen; Crescenzo, Claudia; Guglielmino, Antonino

    2013-01-01

    Purpose Embryo kinetics analysis is an emerging tool for selecting embryo(s) for transfer. The aim of the present study was to determine morphokinetic parameters easily usable in the laboratory and predictive of embryo development and, most importantly, of embryo competence in producing a clinical pregnancy after day 5 transfer. Methods A retrospective time-lapse monitoring analysis of morphokinetic parameters for 72 fully implanted embryos (group A) were compared to 106 non-implanted embryos...

  18. A Simple Method for Transportation of Mouse Embryos Using Microtubes and a Warm Box.

    Directory of Open Access Journals (Sweden)

    Mikiko Tokoro

    Full Text Available Generally, transportation of preimplantation embryos without freezing requires incubators that can maintain an optimal culture environment with a suitable gas phase, temperature, and humidity. Such incubators are expensive to transport. We reported previously that normal offspring were obtained when the gas phase and temperature could be maintained during transportation. However, that system used plastic dishes for embryo culture and is unsuitable for long-distance transport of live embryos. Here, we developed a simple low-cost embryo transportation system. Instead of plastic dishes, several types of microtubes-usually used for molecular analysis-were tested for embryo culture. When they were washed and attached to a gas-permeable film, the rate of embryo development from the 1-cell to blastocyst stage was more than 90%. The quality of these blastocysts and the rate of full-term development after embryo transfer to recipient female mice were similar to those of a dish-cultured control group. Next, we developed a small warm box powered by a battery instead of mains power, which could maintain an optimal temperature for embryo development during transport. When 1-cell embryos derived from BDF1, C57BL/6, C3H/He and ICR mouse strains were transported by a parcel-delivery service over 3 days using microtubes and the box, they developed to blastocysts with rates similar to controls. After the embryos had been transferred into recipient female mice, healthy offspring were obtained without any losses except for the C3H/He strain. Thus, transport of mouse embryos is possible using this very simple method, which might prove useful in the field of reproductive medicine.

  19. Transfer Function of Multi-Stage Active Filters: A Solution Based on Pascal's Triangle and a General Expression

    Science.gov (United States)

    Levesque, Luc

    2012-01-01

    A method is proposed to simplify analytical computations of the transfer function for electrical circuit filters, which are made from repetitive identical stages. A method based on the construction of Pascal's triangle is introduced and then a general solution from two initial conditions is provided for the repetitive identical stage. The present…

  20. Pregnancy rates and serum 13,14-dihydro-15-keto-PGF2α concentrations in recipient Nelore heifers treated with meloxicam after the transfer of in vitro-produced embryos.

    Science.gov (United States)

    Lopes, Larissa Márcia Jonasson; Balbinot, Matheus; Fonseca, Bruna Alves; de Araújo, Cláudio Vieira; Martins, Lílian Rigatto

    2015-09-01

    This study aimed to determine if the administration of meloxicam, a cyclooxygenase 2 inhibitor, to Nelore (Bos taurus indicus) heifers, in which embryo transfer is more difficult and requires a greater manipulation of the reproductive tract than in Bos taurus females, would improve the pregnancy rates by decreasing serum PGFM concentrations. After estrous synchronization, multiparous recipient heifers (n = 85) were selected as embryo recipients and were randomly allocated into two groups: CON (n = 42), the control group, in which animals received 10 mL of saline intramuscularly (the same volume of meloxicam), and MEL (n = 43), the group in which animals were treated with meloxicam. According to the degree of passing the catheter, recipients from both groups were classified as grade I (easy; 80 seconds). One hour before embryo transfer, MEL recipients received an injection of 200 mg of meloxicam. Blood samples were collected from all heifers 1 hour before embryo transfer and 4 and 8 hours after embryo transfer to determine the serum concentrations of 13,14-dihydro-15-keto-PGF2α (PGFM), a PGF2α metabolite. A significant difference in the pregnancy rates on Day 30 was observed in MEL animals between those classified as grade I and II (90.48% vs. 54.54%, respectively; P animals from CON, the pregnancy rates were similar between grades I and II. Serum concentrations of PGFM from samples collected 4 and 8 hours after embryo transfer were lower in pregnant animals from MEL grade I than in pregnant animals from MEL grade II. Considering the pregnant females from CON, no difference was observed from samples collected 4 and 8 hours after embryo transfer. Interestingly, no difference in PGFM serum concentrations was observed between the pregnant females from MEL grade II and pregnant females from CON (P meloxicam had a positive effect on the pregnancy rates of grade I Nelore heifers. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Analysis of Unsteady Tip and Endwall Heat Transfer in a Highly Loaded Transonic Turbine Stage

    Science.gov (United States)

    Shyam, Vikram; Ameri, Ali; Chen, Jen-Ping

    2010-01-01

    In a previous study, vane-rotor shock interactions and heat transfer on the rotor blade of a highly loaded transonic turbine stage were simulated. The geometry consists of a high pressure turbine vane and downstream rotor blade. This study focuses on the physics of flow and heat transfer in the rotor tip, casing and hub regions. The simulation was performed using the Unsteady Reynolds-Averaged Navier-Stokes (URANS) code MSU-TURBO. A low Reynolds number k-epsilon model was utilized to model turbulence. The rotor blade in question has a tip gap height of 2.1 percent of the blade height. The Reynolds number of the flow is approximately 3x10(exp 6) per meter. Unsteadiness was observed at the tip surface that results in intermittent "hot spots". It is demonstrated that unsteadiness in the tip gap is governed by inviscid effects due to high speed flow and is not strongly dependent on pressure ratio across the tip gap contrary to published observations that have primarily dealt with subsonic tip flows. The high relative Mach numbers in the tip gap lead to a choking of the leakage flow that translates to a relative attenuation of losses at higher loading. The efficacy of new tip geometry is discussed to minimize heat flux at the tip while maintaining choked conditions. In addition, an explanation is provided that shows the mechanism behind the rise in stagnation temperature on the casing to values above the absolute total temperature at the inlet. It is concluded that even in steady mode, work transfer to the near tip fluid occurs due to relative shearing by the casing. This is believed to be the first such explanation of the work transfer phenomenon in the open literature. The difference in pattern between steady and time-averaged heat flux at the hub is also explained.

  2. Nucleoli from two-cell embryos support the development of enucleolated germinal vesicle oocytes in the pig.

    Science.gov (United States)

    Kyogoku, Hirohisa; Ogushi, Sugako; Miyano, Takashi

    2012-11-01

    Recent research has shown that nucleoli of oocytes at the germinal vesicle (GV) stage (GV nucleoli) are not necessary for oocyte maturation but are essential for early embryonic development. Nucleoli of 2-cell embryos (2-cell nucleoli) have morphology similar to that of nucleoli in oocytes at the GV stage. In this study, we examined the ability of 2-cell nucleoli to substitute for GV nucleoli in terms of supporting early embryonic development by nucleolus aspiration (enucleolation) and transfer into metaphase II (MII) oocytes or 2-cell embryos that were derived from enucleolated oocytes at the GV stage in the pig. When 2-cell embryos were centrifuged to move the lipid droplets to one side of the blastomere, multiple nucleoli in the nucleus fused into a single nucleolus. The nucleoli were then aspirated from the 2-cell embryos by micromanipulation. The injection of 2-cell nucleoli to GV enucleolated oocytes at the MII stage rescued the embryos from the early embryonic arrest, and the resulting oocytes developed to blastocysts. However, the injection of 2-cell and GV nucleoli to 2-cell embryos derived from GV enucleolated oocytes rarely restored the development to blastocysts. These results indicate that 2-cell nucleoli support early embryonic development as GV nucleoli and that the presence of nucleoli is essential for pig embryos before the 2-cell stage.

  3. Action of uranium on pre implanted mouse embryos

    International Nuclear Information System (INIS)

    Kundt, Miriam S.

    2001-01-01

    The cultured preimplantation embryos are normally employed to evaluate the effects of environmental pollutants specially metals. Embryos were obtained from hybrid females CBA x C57 Bl following induction of super ovulation. They were incubated from 1 cell stage during 120 hs. in M16 cultured medium. Three different experiments were carried out: A, B and C using uranyl nitrate UO 2 (NO 3 ) 2 6H 2 O as source of uranium. In experiment 'A' the embryos were cultivated in the same culture dish containing final U concentrations of 13, 26, 52, 104 and 208 μgU/ml. In experiment 'B' embryos in a one cell stage were placed in culture medium with uranyl nitrate with final U concentrations of 26, 52, 104 μgU/ml. After 24 hours those embryos which had reached the two-cell stage were transferred to another culture dish to which fresh solutions of uranyl nitrate were added, maintaining the same concentrations of the previous one. In experiment 'C' the embryos were cultivated containing final U concentrations of 26, 52 and 104 μgU/ml and they were transferred to another culture dish every day to which fresh solutions of uranyl nitrate were added. Different embryos parameters were analyzed: 1) Development grade; 2) Number of cell per embryo and metaphases index; and 3) Embryo ploidy. 1) Embryos were observed each 24 hs. to evaluate development grade: 2, 4 and 8 cell stage, morula, early -expanded- hatched blastocysts and atresic embryos. No significant differences were observed in the proportion of embryos arrested either in the one-cell or in the two cell stages in control culture medium regarding different concentrations of U, in a total of 4388 embryos analyzed. From 2 cell stage, moment that the embryo begins to synthesize its own ARNm, the delay in embryonic development increased dose dependent. On the other hand, the toxicological effects in the same concentration are increase from 'A' treatment to 'C' treatment. Embriotoxicology effects are evidenced by an increment in

  4. Influence of metal(loid) bioaccumulation and maternal transfer on embryo-larval development in fish exposed to a major coal ash spill.

    Science.gov (United States)

    Greeley, Mark S; Adams, S Marshall; Elmore, Logan R; McCracken, Mary K

    2016-04-01

    In December 2008, an earthen retaining wall at the Tennessee Valley Authority (TVA) Kingston Fossil Fuel Plant failed and released 4.1 millionm(3) of coal ash to rivers flowing into Watts Bar Reservoir in east Tennessee, United States (U.S.). As part of a comprehensive effort to evaluate the risks to aquatic resources from this spill - the largest in U.S. history - we compared bioaccumulation and maternal transfer of selenium (Se), arsenic (As), and mercury (Hg) in adult redear sunfish (Lepomis macrolophus), collected two years after the spill from both coal-ash exposed and non-exposed areas of the Emory and Clinch Rivers, with the success of embryo-larval development in their offspring. Whole body and ovary concentrations of Se in female sunfish at three study sites downstream of the spill were significantly elevated (site means=4.9-5.3 and 6.7-9.0mg/kg d.w. whole body and ovary concentrations, respectively) compared with concentrations in fish from reference sites upstream of the spill site (2.2-3.2mg/kg d.w. for whole bodies and 3.6-4.8mg/kg d.w. for ovaries). However, Se concentrations in coal ash-exposed areas remain below proposed U.S. Environmental Protection Agency (USEPA) criteria for the protection of aquatic life. Site-to-site variation in fish concentrations of As and Hg were not well-correlated with ash-exposure, reflecting the multiple sources of these metal(loid)s in the affected watersheds. In 7-day laboratory tests of embryos and larvae derived from in vitro crosses of eggs and sperm from these field-collected sunfish, fertilization success, hatching success, embryo-larval survival, and incidences of developmental abnormalities did not differ significantly between ash-exposed and non-exposed fish. Furthermore, these developmental endpoints were not correlated with whole body or ovary concentrations of Se, As, or Hg in the maternal fish, or with fish size, ovary weight, or gonadal-somatic indices. Results from this and related studies associated

  5. The Influence of Nutrient Availability on Trophic Energy Transfer Using Two-Stage Continuous Cultures

    Science.gov (United States)

    McBeain, K. A.; Halsey, K.

    2016-02-01

    Areas of extreme nutrient limitation are expected to expand in the surface ocean as a consequence of climate change. Phytoplankton are the base of the marine food web and their physiology is strongly influenced by nutrient availability. Two-stage continuous cultures were used to investigate trophic energy and nutrient transfers between phytoplankton and microzooplankton under conditions simulating high and low nutrient availability. The first stage contained either fast (μ=1.2 d-1) or slow (μ=0.2 d-1) growing nitrogen-limited prey organisms: a green alga (Dunaliella tertiolecta) or a diatom (Thalassiosira pseudonana). The second stage contained the dinoflagellate Oxyrrhis marina, which was fed a constant biomass of prey delivered from the first stage. Cultures were allowed to reach steady state before the physiologies of both prey and predator were analyzed. The population of O. marina was five-fold higher when fed with the fast growing D. tertiolecta compared to slow-growing D. tertiolecta. D. tertiolecta also supported a population of O. marina that was an order of magnitude higher than the diatom prey type. The fitness of the prey and O. marina under each nutrient limiting condition were further assessed using a suite of physiological measurements including chlorophyll, C:N:P ratios, lipid body formation, cell volume, motility, and fecal pellet accumulation. Regardless of prey type, O. marina were consistently larger and less abundant when fed prey growing at 0.2 d-1. These results suggest that major shifts in ecosystem food web structure are likely as surface waters become increasingly depleted of nutrients.

  6. Effect of two activation treatments and age of blastomere karyoplasts on in vitro development of bovine nuclear transfer embryos

    DEFF Research Database (Denmark)

    Booth, P J; Holm, P; Vajta, G

    2001-01-01

    The yield and quality of (a) parthenogenetic blastocysts produced by two activation treatments (cycloheximide [CHX] or 6-dimethylaminopurine [DMAP]) and (b) nuclear transfer blastocysts generated using these two activation treatments and three different ages of karyoplast derived from day 3, 4, o...

  7. Effect of vitamin A supplementation at different gaseous environments on in vitro development of pre-implantation sheep embryos to the blastocyst stage.

    Science.gov (United States)

    Rajesh, N; Shankar, M B; Deecaraman, M

    2010-11-01

    Vitamin A (all-trans retinol) is an important antioxidant whose role in embryo development in vitro and in vivo is well established. Oxidative stress is a major cause of defective embryo development. This study evaluated the effects of all-trans retinol supplementation to maturation and embryo culture media under different gaseous environments on the development of ovine oocytes and embryos in vitro. The percentages of cleavage, morula and blastocyst, total cell count and comet assay were taken as indicators of developmental competence of embryos. In experiments I and II, all-trans retinol at concentrations of 0, 2, 4, 6, 8 and 10 μM were supplemented to the oocyte maturation medium and cultured in an environment of 5% or 20% O2 respectively. All-trans retinol supplementation (6 μM) to the maturation medium at 5% O2 levels significantly increased blastocyst yield and total cell number (P embryo culture media under a 5% or 20% O2 environment, respectively. All-trans retinol supplementation to the embryo culture medium at 5% O2 levels did not yield any significant result whereas the culture at 20% O2 levels gave significantly higher blastocyst yield in the 6 μM supplemented group compared with the control group (P < 0.01).

  8. Aplicação da ultrassonografia colorida doppler em programas de transferência de embriões equinos Potential use of doppler ultrasound in equine embryo transfer programs

    Directory of Open Access Journals (Sweden)

    Jair Camargo Ferreira

    2011-06-01

    distúrbios de fluxo sanguíneo de trato reprodutivo ainda se fazem necessários.Embryo transfer in mares has been commercially used for more than three decades and it is one of most frequently applied biotechnologies on equine reproduction. Ultrasonic doppler exam of donors during the pre and post-breeding and recipients evaluation at the embryo transfer moment is essential for embryo transfer program success. Doppler ultrasonography is a non-invasive technic that allows real-time evaluation of the reproductive system hemodynamics in large animal. For providing anatomical details and blood-flow physiologic information of vessels and tissues, doppler exam can be used for ovulation prognostication, and to evaluate uterine and luteal functional status. Additionally, it can serve as a diagnostic aid of hemodynamic disturbance in reproductive system. Vascularity changes of future ovulatory follicles precede the diameter deviation. Based on this, it is possible to estimate the beginning of the breeding season and the best moment to initiate superovulatory treatments. Moreover, follicular blood flow can be used to decide the most appropriated moment for ovulation induction and breeding of donors mares. Uterine and luteal evaluation using Doppler-mode in mares can be useful to select embryo recipients with satisfactory progesterone production and adequate uterine vascularity for embryo development and maintenance of pregnancy. The maternal recognition of pregnancy also can be evaluated. Although the applicability of the Doppler technology in embryo transfer programs, additional studies are necessary to determine the standards of normality and to characterize different blood-flow disturbances of the reproductive system.

  9. Maternal hCG concentrations in early IVF pregnancies: associations with number of cells in the Day 2 embryo and oocytes retrieved.

    Science.gov (United States)

    Tanbo, T G; Eskild, A

    2015-12-01

    Do number of cells in the transferred cleavage stage embryo and number of oocytes retrieved for IVF influence maternal hCG concentrations in early pregnancies? Compared with transfer of a 2-cell embryo, transfer of a 4-cell embryo results in higher hCG concentrations on Day 12 after transfer, and more than 20 oocytes retrieved were associated with low hCG concentrations. Maternal hCG concentration in very early pregnancy varies considerably among women, but is likely to be an indicator of time since implantation of the embryo into the endometrium, in addition to number and function of trophoblast cells. We followed 1047 pregnancies after IVF/ICSI from oocyte retrieval until Day 12 after embryo transfer. Women were recruited in Norway during the years 2005-2013. Successful pregnancies after transfer of one single embryo that had been cultured for 2 days were included. Maternal hCG was quantified on Day 12 after embryo transfer by chemiluminescence immunoassay, which measures intact hCG and the free β-hCG chain. Information on a successful pregnancy, defined as birth after >16 weeks, was obtained by linkage to the Medical Birth Registry of Norway. Transfer of a 4-cell embryo resulted in higher maternal hCG concentrations compared with transfer of a 2-cell embryo (134.8 versus 87.8 IU/l, P 20) was associated with low hCG concentrations (P hCG concentrations in early pregnancy. Although embryo transfer was performed at the same time after fertilization, we do not know the exact time of implantation. A further limitation to our study is that the number of pregnancies after transfer of a 2-cell embryo was small (27 cases). Number of cells in the transferred embryo and number of oocytes retrieved may influence the conditions and timing for embryo implantation in different ways and thereby influence maternal hCG concentrations. Such knowledge may be important for interpretation of hCG concentrations in early pregnancy. © The Author 2015. Published by Oxford University

  10. Rabbit whole embryo culture.

    Science.gov (United States)

    Marshall, Valerie A; Carney, Edward W

    2012-01-01

    Although the rabbit is used extensively in developmental toxicity testing, relatively little is known about the fundamental developmental biology of this species let alone mechanisms underlying developmental toxicity. This paucity of information about the rabbit is partly due to the historic lack of whole embryo culture (WEC) methods for the rabbit, which have only been made available fairly recently. In rabbit WEC, early somite stage embryos (gestation day 9) enclosed within an intact amnion and attached to the visceral yolk sac are dissected from maternal tissues and placed in culture for up to 48 h at approximately 37°C and are continuously exposed to an humidified gas atmosphere mixture in a rotating culture system. During this 48 h culture period, major phases of organogenesis can be studied including cardiac looping and segmentation, neural tube closure, and development of anlagen of the otic system, eyes and craniofacial structures, somites and early phases of limb development (up to bud stage), as well as expansion and closure of the visceral yolk sac around the embryo. Following completion of the culture period, embryos are evaluated based on several growth and development parameters and also are assessed for morphological abnormalities. The ability to sustain embryo development independent of the maternal system allows for exposure at precise development stages providing the opportunity study the direct action of a teratogen or one of its metabolites on the developing embryo. Rabbit WEC is perhaps most useful when used in conjunction with rodent WEC methods to investigate species-specific mechanisms of developmental toxicity.

  11. Evidence for stage-specific modulation of specific microRNAs (miRNAs) and miRNA processing components in zygotic embryo and female gametophyte of loblolly pine (Pinus taeda).

    Science.gov (United States)

    Oh, Thomas J; Wartell, Roger M; Cairney, John; Pullman, Gerald S

    2008-01-01

    MicroRNAs (miRNAs) are known to regulate plant development, but have not been studied in gymnosperm seed tissues. The presence and characteristics of several miRNAs were examined in zygotic embryos (ZEs) and female gametophytes (FGs) of Pinus taeda (loblolly pine). Evidence for miRNAs was obtained using northern analyses and quantitative reverse transcription polymerase chain reaction (qRT-PCR) mediated with poly(A) polymerase. Partial sequences of two miRNAs were verified. Three regions of putative mRNA targets were analyzed by qRT-PCR to monitor the occurrence of stage-dependent miRNA-mediated cleavage. Five miRNAs were identified in ZEs and FGs along with partial sequences of Pta-miR166 and Pta-miR167. Both miRNAs showed differing degrees of tissue-specific and stage-specific modulation. Analysis of HB15L mRNA (a potential Pta-miR166 target) suggested miRNA-guided cleavage in ZEs and FGs. Analysis of ARF8L mRNA (a potential Pta-miR167 target) implied cleavage in ZEs but not in FGs. Argonaute9-like mRNA (ptAGO9L) showed stage-specific modulation of expression in ZEs that appeared to be inverted in the corresponding FGs. MicroRNAs and argonaute genes varied spatiotemporally during seed development. The peak levels of Pta-miR166 in FGs and ptAGO9L in embryos occurred at stage 9.1, a critical transition point during embryo development and a point where somatic embryo maturation often stops. MicroRNAs identified in FG tissue may play a role in embryogenesis.

  12. Effect of Levothyroxine on Miscarriage Among Women With Normal Thyroid Function and Thyroid Autoimmunity Undergoing In Vitro Fertilization and Embryo Transfer: A Randomized Clinical Trial.

    Science.gov (United States)

    Wang, Haining; Gao, Hongwei; Chi, Hongbin; Zeng, Lin; Xiao, Wenhua; Wang, Yanrong; Li, Rong; Liu, Ping; Wang, Chen; Tian, Qing; Zhou, Zehong; Yang, Jin; Liu, Ye; Wei, Rui; Mol, Ben Willem J; Hong, Tianpei; Qiao, Jie

    2017-12-12

    Presence of thyroid autoantibodies in women with normal thyroid function is associated with increased risk of miscarriage. Whether levothyroxine treatment improves pregnancy outcomes among women undergoing in vitro fertilization and embryo transfer (IVF-ET) is unknown. To determine the effect of levothyroxine on miscarriage among women undergoing IVF-ET who had normal thyroid function and tested positive for thyroid autoantibodies. An open-label, randomized clinical trial involving 600 women who tested positive for the antithyroperoxidase antibody and were being treated for infertility at Peking University Third Hospital from September 2012 to March 2017. The intervention group (n = 300) received either a 25-μg/d or 50-μg/d dose of levothyroxine at study initiation that was titrated according to the level of thyroid-stimulating hormone during pregnancy. The women in the control group (n = 300) did not receive levothyroxine. All participants received the same IVF-ET and follow-up protocols. The primary outcome was the miscarriage rate (pregnancy loss before 28 weeks of gestation, which was calculated among women who became pregnant). The secondary outcomes were clinical intrauterine pregnancy rate (fetal cardiac activity seen at sonography observation on the 30th day after the embryo transfer), and live-birth rate (at least 1 live birth after 28 weeks of gestation). Among the 600 women (mean [SD] age, 31.6 [3.8] years) randomized in this trial, 567 women (94.5%) underwent IVF-ET and 565 (94.2%) completed the study. Miscarriage rates were 10.3% (11 of 107) in the intervention group and 10.6% (12 of 113) in the control group, with the absolute rate difference (RD) of -0.34% (95% CI, -8.65% to 8.12%) over the 4.5-year study period. Clinical intrauterine pregnancy rates were 35.7% (107 of 300) in the intervention group and 37.7% (113 of 300) in the control group, with an absolute RD of -2.00% (95% CI, -9.65% to 5.69%). Live-birth rates were 31.7% (95 of 300

  13. A two-stage biological gas to liquid transfer process to convert carbon dioxide into bioplastic

    KAUST Repository

    Al Rowaihi, Israa

    2018-03-06

    The fermentation of carbon dioxide (CO2) with hydrogen (H2) uses available low-cost gases to synthesis acetic acid. Here, we present a two-stage biological process that allows the gas to liquid transfer (Bio-GTL) of CO2 into the biopolymer polyhydroxybutyrate (PHB). Using the same medium in both stages, first, acetic acid is produced (3.2 g L−1) by Acetobacterium woodii from 5.2 L gas-mixture of CO2:H2 (15:85 v/v) under elevated pressure (≥2.0 bar) to increase H2-solubility in water. Second, acetic acid is converted to PHB (3 g L−1 acetate into 0.5 g L−1 PHB) by Ralstonia eutropha H16. The efficiencies and space-time yields were evaluated, and our data show the conversion of CO2 into PHB with a 33.3% microbial cell content (percentage of the ratio of PHB concentration to cell concentration) after 217 h. Collectively, our results provide a resourceful platform for future optimization and commercialization of a Bio-GTL for PHB production.

  14. Office microlaparoscopic intrafallopian transfer of day one zygote ...

    African Journals Online (AJOL)

    The objective of the study was to investigate whether transferring zygotes on day 1 would result in similar pregnancy rates compared to transferring cleavage stage embryos on day 3 in a prospective randomized trial, using the office microlaparoscopic procedure. Patients undergoing IVF/ICSI treatments were randomized to ...

  15. Optimization of a novel nylon mesh container for human embryo ultrarapid vitrification.

    Science.gov (United States)

    Nakashima, Akira; Ino, Nao; Kusumi, Maki; Ohgi, Shirei; Ito, Megumu; Horikawa, Takashi; Nakagawa, Koji; Saito, Takakazu; Kamura, Toshiharu; Saito, Hidekazu

    2010-05-01

    To evaluate the efficacy of a nylon mesh container in vitrification of human embryos and to determine the optimal osmotic pressure of the initial thawing solution. Retrospective analysis. National Center for Child Health and Development, Tokyo, Japan. Infertile patients undergoing either in vitro fertilization or intracytoplasmic sperm injection in our hospital. Embryos, at the cleavage stage, were cryopreserved using the vitrification method in either a plastic straw or a nylon mesh container. The embryos were thawed using an initial osmotic pressure of either 0.5 M or 1.0 M sucrose with subsequent step-wise dilution. After thawing, the embryos were transferred to the uterus. Survival rate of blastomeres, embryo survival rate, implantation, and pregnancy rates, cancellation rate because of embryo damage. Use of nylon mesh and the 1.0 M sucrose thawing solution significantly improved blastomere survival rate (98.0 +/- 1.0%, mean +/- SEM), pregnancy rate (41.0%) and implantation rate (32.3%). Vitrification using a nylon mesh container and subsequent thawing in a 1.0 M sucrose solution is an easy and inexpensive method that improves the reliability of embryo cryopreservation of embryos without adverse effects on clinical outcomes. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  16. Near-infrared laser irradiation improves the development of mouse pre-implantation embryos.

    Science.gov (United States)

    Yokoo, Masaki; Mori, Miho

    2017-05-27

    The aim of the present study was to assess the effects of near-infrared laser irradiation on the in vitro development of mouse embryos. Female ICR mice were superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin (hCG), and mated with male mice. Two-cell stage embryos were collected 40 h after administering hCG and cultured in M16 medium. Two-cell embryos (0 h after culture), 8-cell embryos (approx. 30 h after culture), morula (approx. 48 h after culture), and blastocysts (approx. 73 h after culture) were irradiated at 904 nm for 60 s. These embryos were cultured in a time-lapse monitoring system and the timing of blastocyst hatching was evaluated. Some of the irradiated blastocysts were transferred to the uterine horns of pseudopregnant recipients immediately after irradiation. Pregnancy rates, and offspring growth and fertility, were evaluated. Near-infrared laser irradiation increased the speed of in vitro mouse embryo development. In irradiated blastocysts, hatching was faster than in control (non-irradiated) blastocysts (18.4 vs. 28.2 h, P embryo development in vitro, and increases the live birth rate without affecting the normality of the offspring. Thus, the near-infrared laser method may enhance the quality of embryos and contribute to improvements in reproductive technologies in mammals. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Modified hMG stimulated: an effective option in endometrial preparation for frozen-thawed embryo transfer in patients with normal menstrual cycles.

    Science.gov (United States)

    Huang, Pinxiu; Wei, Lihong; Li, Xinlin; Lin, Zhong

    2018-04-20

    To evaluate the clinical efficacy of modified human menopausal gonadotropin (hMG) stimulated, hormone replacement therapy (HRT), natural cycling and letrozole ovulation induction during endometrial preparation for frozen-thawed embryo transfer (FET) in patients with normal menstrual cycles. This retrospective analysis included a total of 5070 cycles of patients with normal menstrual patterns who underwent FET between October 2009 and September 2015. The patients were divided into four groups according to the method of endometrial preparation for FET: 1838 cycles were natural, 1666 underwent HRT, 340 underwent letrozole ovulation induction and 1226 underwent modified hMG stimulated. Reproduction-related clinical outcomes in the four groups were compared. The clinical pregnancy rates and live birth rates of patients in the modified hMG stimulated group were significantly higher than that in the other groups p .05). Modified hMG stimulated resulted in a higher pregnancy rate compared to the other treatment groups. Therefore, modified hMG stimulated may be an effective option in endometrial preparation for FET in patients with normal menstrual cycles.

  18. Obligatory versus elective single embryo transfer in in vitro fertilization. A population-based analysis of data from the U.K. Human Fertilisation and Embryology Authority.

    Science.gov (United States)

    Straughen, Jennifer K; Salihu, Hamisu M; Keith, Louis; Petrozzino, Jeffrey; Jones, Christopher

    2013-01-01

    To determine how obligatory single embryo transfer (SET) and elective SET influence pregnancy outcome. We compared women who underwent obligatory and elective SET using data from a comprehensive, population-based register from the United Kingdom Human Fertilisation and Embryology Authority, which contained all in vitro fertilization (IVF) treatments administered between 1991 and 1998. Generalized estimating equations were used to generate adjusted odds ratios (ORs) and 95% confidence intervals (CIs) to compare clinical pregnancy, live birth, and multiple birth rates. Obligatory and elective SET had similar clinical pregnancy and live birth rates and comparable multiple birth rates. Obligatory and elective SET were equally likely to end in a live birth (OR = 1.08; 95% CI = 0.90, 1.30). Similar results were found after restricting the data to women without previous IVF births (OR = 1.18; 95% CI = 0.98, 1.42) and without previous naturally conceived live births (OR = 1.16; 95% CI = 0.95, 1.43). This study suggests that obligatory SET can achieve pregnancy and live birth rates that are at least as good as elective SET. Equally important is the low multiple birth rate which was maintained in both forms of SET. More studies comparing elective versus obligatory SET can assist with achieving optimal pregnancy rates while preventing multiple births.

  19. Laparoscopic cholecystectomy due to acute calculouscholecystitis in 16 weeks′ in vitro fertilization and embryo transfer pregnancy: Report of the first case

    Directory of Open Access Journals (Sweden)

    G Augustin

    2012-01-01

    Full Text Available The most common casues of acute abdomen during pregnancy are acute appendicitis followed by acute cholecystitis. The case presented is a 33-year-old patient in 16 weeks′ in vitro fertilization and embryo transfer pregnacy who developed acute cholecystitis. Previously there were two unsuccessful cycles, one complicated with ovarian hyperstimulation syndrome. Due to clinical deterioration during intravenous antibiotic therapy laparoscopic cheolecystecomy was performed and acute cholecystitis found. The postoperative course was uneventful. During the first 24 h tocolysis with intravenous fenoterol in addition to peroral atenolol 2 Χ 50 mg was administered. Postoperative course was uneventuful with further normal pregnancy. Elective cesarean section was made in term pregnancy (39 weeks with singleton with Apgar 10/10. Current guidelines do not recommend prophylactic tocolysis in pregnant population with acute abdomen but there is no mention of the IVF-ET subpopulation of patients. Also, there are no guidelines for thromboprophylaxis in such patients with increased risk of thromboembolic accidents. To our knowledge this is the first case report of a laparoscopic cholecystectomy during IVF-ET gestation.

  20. [Limitations and controversies in determining the predictive value of oocyte and embryo morphology criteria].

    Science.gov (United States)

    Figueira, Rita de Cássia Savio; Aoki, Tsutomu; Borges Junior, Edson

    2015-11-01

    In order to increase the success rate of in vitro fertilization cycles, several studies have focused on the identification of the embryo with higher implantation potential. Despite recent advances in the reproductive medicine, based on the OMICs technology, routinely applicable methodologies are still needed. Thus, in most fertilization centers embryo selection for transfer is still based on morphological parameters evaluated under light microscopy. Several morphological parameters may be evaluated, ranging from the pronuclear to blastocyst stage. In general, despite the day of transfer, some criteria are suggested to present a predictive value for embryo viability when analyzed independently or combined. However, the subjectivity of morphological evaluation, as well as the wide diversity of embryo classification systems used by different fertilization centers shows contrasting results, making the implementation of a consensus regarding different morphological criteria and their predictive value a difficult task. The optimization of embryo selection represents a large potential to increase treatment success rates, allowing the transfer of a reduced number of embryos and minimizing the risks of multiple pregnancy.

  1. Contrasting Storage Protein Synthesis and Messenger RNA Accumulation during Development of Zygotic and Somatic Embryos of Alfalfa (Medicago sativa L.) 1

    Science.gov (United States)

    Krochko, Joan E.; Pramanik, Saroj K.; Bewley, J. Derek

    1992-01-01

    During development on hormone-free media, somatic embryos pass through distinct morphological stages that superficially resemble those of zygotic embryo development (globular, heart, torpedo, cotyledonary stages). Despite these similarities, they differ from zygotic embryos in the extent of cotyledonary development and the patterns of synthesis and quantitative expression of seed-specific storage proteins (7S, 11S, and 2S proteins). Alfin (7S) is the first storage protein synthesized in developing zygotic embryos (stage IV). The 11S (medicagin) and 2S (Low Molecular Weight, LMW) storage proteins are not detectable until the following stage of development (stage V), although all three are present before the completion of embryo enlargement. Likewise, the 7S storage protein is the first to be synthesized in developing somatic embryos (day 5). Medicagin is evident by day 7 and the LMW protein by day 10. In contrast to zygotic embryos, alfin remains the predominant storage protein in somatic embryos throughout development. Not only are the relative amounts of medicagin and the LMW protein reduced in somatic embryos but the LMW protein is accumulated much later than the other proteins. Quantification of the storage protein mRNAs (7S, 11S, and 2S) by northern blot analysis confirms that there are substantial differences in the patterns of message accumulation in zygotic and somatic embryos of alfalfa (Medicago sativa). In zygotic embryos, the 7S, 11S, and 2S storage protein mRNAs are abundant during maturation and, in particular, during the stages of maximum protein synthesis (alfin, stages VI and VII; medicagin, stage VII; LMW, stage VII). In somatic embryos, the predominance of the 7S storage protein is correlated with increased accumulation of its mRNA, whereas the limited synthesis of the 11S storage protein is associated with much lower steady-state levels of its message. The mRNA for the LMW protein is present already by 3 days after transfer to hormone-free media

  2. Aberrant epigenetic reprogramming of imprinted microRNA-127 and Rtl1 in cloned mouse embryos

    International Nuclear Information System (INIS)

    Cui Xiangshun; Zhang Dingxiao; Ko, Yoeung-Gyu; Kim, Nam-Hyung

    2009-01-01

    The microRNA (miRNA) genes mir-127 and mir-136 are located near two CpG islands in the imprinted mouse retrotransposon-like gene Rtl1, a key gene involved in placenta formation. These miRNAs appear to be involved in regulating the imprinting of Rtl1. To obtain insights into the epigenetic reprogramming of cloned embryos, we compared the expression levels of mir-127 and mir-136 in fertilized mouse embryos, parthenotes, androgenotes and cloned embryos developing in vitro. We also examined the DNA methylation status of the promoter regions of Rtl1 and mir-127 in these embryos. Our data showed that mir-127 and mir-136 were highly expressed in parthenotes, but rarely expressed in androgenotes. Interestingly, the expression levels of mir-127 and mir-136 in parthenotes were almost twice that seen in the fertilized embryos, but were much lower in the cloned embryos. The Rtl1 promoter region was hyper-methylated in blastocyst stage parthenotes (75.0%), moderately methylated (32.4%) in the fertilized embryos and methylated to a much lower extent (∼10%) in the cloned embryos. Conversely, the promoter region of mir-127 was hypo-methylated in parthenogenetically activated embryos (0.4%), moderately methylated (30.0%) in fertilized embryos and heavily methylated in cloned blastocysts (63-70%). These data support a role for mir-127 and mir-136 in the epigenetic reprogramming of the Rtl1 imprinting process. Analysis of the aberrant epigenetic reprogramming of mir-127 and Rtl1 in cloned embryos may help to explain the nuclear reprogramming procedures that occur in donor cells following somatic cell nuclear transfer (SCNT).

  3. Early maternal serum ß-human chorionic gonadotropin (ß-hCG) levels and sex-related growth difference of IVF embryos.

    Science.gov (United States)

    Esh-Broder, Efrat; Oron, Galia; Son, Weon-Young; Holzer, Hananel; Tulandi, Togas

    2015-10-01

    Maternal serum ß-human chorionic gonadotropin (ß-hCG) represents the trophoblastic cell mass and is an indirect measurement of embryo development at early implantation stage. Studies in animals and human embryos detected sex-related growth differences (SRGD) in favour of male embryos during the pre-implantation period. The purpose of our study was to correlate SRGD and maternal serum ß-hCG at 16 days after embryo transfer. We retrospectively analysed all (fresh and frozen) non-donor, single embryo transfers (SET), elective and not elective, that were performed between December 2008 and December 2013. We included ß-hCG values from day 16 after oocyte collection of pregnancies resulting in live birth. Neonatal gender was retrieved from patient files. Male and female embryos were further grouped to cleavage and blastocyst stage transfers. Regression analysis for confounding variables included maternal age, maternal body mass index (BMI), use of micromanipulation (ICSI), embryo quality (grade), assisted hatching, day of transfer and fresh or frozen embryo transfer. Seven hundred eighty-six non-donor SETs resulted in live birth. After including only day 16 serum ß-hCG results, 525 SETs were analysed. Neonatal gender was available for 522 cases. Mean maternal serum ß-hCG levels were similar, 347 ± 191 IU/L in the male newborn group and 371 ± 200 IU/L in the female group. The difference between ß-hCG levels remained insignificant after adjusting for confounding variables. Early maternal ß-hCG levels after embryo transfers did not represent SRGD in our study.

  4. Acephalous lamb from an in vitro-produced sheep embryo

    OpenAIRE

    Shirazi, Abolfazl; Ahmadi, Ebrahim; Jadidi, Majid; Shams-Esfandabadi, Naser; Heidari, Banafsheh

    2009-01-01

    This is the first report of an acephalous lamb from the transfer of an in vitro-produced sheep embryo. Twelve in vitro-fertilized embryos were transferred to 4 recipient ewes (3 embryos/ewe). Two ewes remained pregnant: one delivered a normal female lamb, the other a male acephalous lamb. Possible contributing factors are discussed.

  5. Prevention of multiple pregnancies in couples with unexplained or mild male subfertility: randomised controlled trial of in vitro fertilisation with single embryo transfer or in vitro fertilisation in modified natural cycle compared with intrauterine insemination with controlled ovarian hyperstimulation.

    Science.gov (United States)

    Bensdorp, A J; Tjon-Kon-Fat, R I; Bossuyt, P M M; Koks, C A M; Oosterhuis, G J E; Hoek, A; Hompes, P G A; Broekmans, F J M; Verhoeve, H R; de Bruin, J P; van Golde, R; Repping, S; Cohlen, B J; Lambers, M D A; van Bommel, P F; Slappendel, E; Perquin, D; Smeenk, J M; Pelinck, M J; Gianotten, J; Hoozemans, D A; Maas, J W M; Eijkemans, M J C; van der Veen, F; Mol, B W J; van Wely, M

    2015-01-09

    To compare the effectiveness of in vitro fertilisation with single embryo transfer or in vitro fertilisation in a modified natural cycle with that of intrauterine insemination with controlled ovarian hyperstimulation in terms of a healthy child. Multicentre, open label, three arm, parallel group, randomised controlled non-inferiority trial. 17 centres in the Netherlands. Couples seeking fertility treatment after at least 12 months of unprotected intercourse, with the female partner aged between 18 and 38 years, an unfavourable prognosis for natural conception, and a diagnosis of unexplained or mild male subfertility. Three cycles of in vitro fertilisation with single embryo transfer (plus subsequent cryocycles), six cycles of in vitro fertilisation in a modified natural cycle, or six cycles of intrauterine insemination with ovarian hyperstimulation within 12 months after randomisation. The primary outcome was birth of a healthy child resulting from a singleton pregnancy conceived within 12 months after randomisation. Secondary outcomes were live birth, clinical pregnancy, ongoing pregnancy, multiple pregnancy, time to pregnancy, complications of pregnancy, and neonatal morbidity and mortality 602 couples were randomly assigned between January 2009 and February 2012; 201 were allocated to in vitro fertilisation with single embryo transfer, 194 to in vitro fertilisation in a modified natural cycle, and 207 to intrauterine insemination with controlled ovarian hyperstimulation. Birth of a healthy child occurred in 104 (52%) couples in the in vitro fertilisation with single embryo transfer group, 83 (43%) in the in vitro fertilisation in a modified natural cycle group, and 97 (47%) in the intrauterine insemination with controlled ovarian hyperstimulation group. This corresponds to a risk, relative to intrauterine insemination with ovarian hyperstimulation, of 1.10 (95% confidence interval 0.91 to 1.34) for in vitro fertilisation with single embryo transfer and 0.91 (0

  6. Not all twins are monozygotic after elective single embryo transfer: analysis of 32,600 elective single embryo transfer cycles as reported to the Society for Assisted Reproductive Technology.

    Science.gov (United States)

    Vega, Mario; Zaghi, Sahar; Buyuk, Erkan; Jindal, Sangita

    2018-01-01

    To study the incidence and risk factors of multiple pregnancies after elective single ET. Historic cohort. Not applicable. Women single ET entered in the SART CORS database from 2010 to 2013. Not applicable. Rate of sex discordant pregnancies. Rate of same sex pregnancies and risk factors for both same sex and sex discordant pregnancies. A total of 32,600 cycles were reported to SART CORS during this time period. There were 15,143 pregnancies from which 14,888 were singletons (98.3%), 23 sex discordant (0.15%) multiple pregnancies, 226 (1.5%) sex concordant multiple pregnancies, and 6 (0.01%) pregnancies without sex information. When Weinberg's differential rule was applied, the rate of dizygotic pregnancies was calculated to be 18%. Unexplained infertility was found to be the biggest risk factor for sex discordant multiple pregnancies (adjusted odds ratio 4.33, 95% confidence interval 1.4-13.1), followed by elevated body mass index (BMI). The only risk factor found for sex concordant pregnancies was undergoing a fresh transfer (adjusted odds ratio 1.4, 95% confidence interval 1.02-1.95). Elective single ET improves, but does not completely eliminate the risk of multiple pregnancies. Patients should be counseled that there might be up to a ∼2% risk of multiple pregnancies, of which up to 18% can be dizygotic. Patients with elevated BMI and unexplained fertility may be at higher risk for sex discordant multiple pregnancies and patients undergoing fresh cycles may be at higher risk for sex concordant multiple pregnancies. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  7. Cryopreservation of green fluorescent protein (GFP)-labeled primordial germ cells with GFP fused to the 3' untranslated region of the nanos gene by vitrification of Japanese eel (Anguilla japonica) somite stage embryos.

    Science.gov (United States)

    Kawakami, Y; Ishihara, M; Saito, T; Fujimoto, T; Adachi, S; Arai, K; Yamaha, E

    2012-12-01

    Primordial germ cells (PGC) are the only cell type in developing embryos with the potential to transmit genetic information to the next generation. In this study, PGC of Japanese eel (Anguilla japonica) were visualized by injection of mRNA synthesized from a construct carrying the green fluorescent protein (GFP) gene fused to the 3' untranslated region of the Japanese eel nanos gene. We investigated the feasibility of cryopreserving Japanese eel PGC by vitrification of dechorionated whole somite stage embryos. The GFP-labeled PGC were rapidly cooled using liquid nitrogen after exposure to a pretreatment solution containing 1.5 M cryoprotectant (methanol, dimethyl sulfoxide, and glycerol for 10 min and ethylene glycol for 10, 20, and 30 min) and a vitrification solution containing 3 M cryoprotectant and 0.5 M sucrose for 1, 5, and 10 min. Ethylene glycerol is an effective cryoprotectant for embryonic cells and shows no evidence of ice formation after thawing. Vitrified and thawed PGC were transplanted into blastula stage embryos from zebrafish (Danio rerio). The GFP-labeled PGC migrated toward the host gonadal ridge, suggesting maintenance of their normal migration motility. These techniques may assist in achieving inter- and intraspecies germ-line chimers using donor Japanese eel PGC.

  8. Cytogenetics and immature embryo culture at Embrapa Trigo breeding program: transfer of disease resistance from related species by artificial resynthesis of hexaploid wheat (Triticum aestivum L. em. Thell

    Directory of Open Access Journals (Sweden)

    Maria Irene Baggio de Moraes Fernandes

    2000-12-01

    to facilitate gene flow between wheat and related species. Since the environment at the center of origin of wheat in Southern Asia is quite different from subtropical environments, Brazilian breeding programs overcome more challenges to adapt wheat crop to biotic and abiotic stresses than some other countries. The germplasm bank of Embrapa Trigo has about 1000 registered entries of Triticum relatives, Aegilops, Secale and Agropyron species supplied from several germplasm banks distributed over the world which were multiplied and/or selected for naturally occurring or artificially inoculated fungal diseases. Since Aegilops squarrosa L. entries showed very good performance, the genetic variability observed in this species was firstly exploited. It is reported here the strategy used for transferring useful genes from Ae. squarrosa (DD, 2n = 14: crossing with tetraploid species (AABB, 2n = 28, rescue and in vitro culture of immature embryos for regeneration of the trihaploid (ABD, 2n = 21 hybrid, and colchicine treatment for genome duplication resulting in the artificial synthesis of hexaploid wheat lines (AABBDD, 2n = 42. Results of 10,739 artificial pollinations involving 28 cross combinations amongst eight T. durum L., T. dicoccum and T. cartlicum tetraploid entries used as female parents and ten selected Ae. squarrosa sources of resistance as male parents are presented here. Immature embryos from 18 cross combinations were recovered and cultured in vitro. Green plantlets from 13 combinations were regenerated. Fertile amphiploids were recovered only from crosses among entries of tetraploid T. durum and diploid Ae. squarrosa. They originated 11 fertile synthetic amphiploid lines from seven different combinations. Useful stem and leaf rust as well as powdery mildew resistance for future use in breeding programs were obtained.

  9. Characterizing the endometrial microbiome by analyzing the ultra-low bacteria from embryo transfer catheter tips in IVF cycles: Next generation sequencing (NGS analysis of the 16S ribosomal gene

    Directory of Open Access Journals (Sweden)

    Xin Tao

    2017-03-01

    Full Text Available There is growing interest in the microbiome of the reproductive tract. The vaginal and placental microbiome has been partially characterized and shown to be related to obstetric outcomes. In this study, we developed a sensitive method to analyze 16S ribosomal RNA profiling from ultra-low bacteria counts, then studied the endometrial microbial environment by analyzing catheter tips after embryo transfers during in vitro fertilization (IVF. An extensive validation was performed on extracted DNA and culture lysates without DNA isolation from the single- or poly-microbial samples of Escherichia coli, Proteus vulgaris, Staphylococcus epidermidis, and Bacillus cereus by Illumina 16S V4 metagenomics workflows. The direct lysis method can reliably detect the genus or species taxonomic level for both single- and poly-microbial samples when there were more than 60 bacterial cells in the culture lysates. Over 99% total operational taxonomic units (OTUs were assigned to correct genus or species. The endometrial microbiome at the time of embryo transfer was characterized by analyzing catheter tips with Illumina V4 metagenomics for 70 patients who underwent IVF cycles. Lactobacillus spp. were detected in all 70 samples. Other vaginal bacteria (Corynebacterium, Bifidobacterium, Staphylococcus, and Streptococcus were also detected. The Illumina V4 metagenomics workflow with preamplification provided a rapid and sensitive method for the identification of bacterial genus or species in single- or poly-microbial samples and clinical embryo transfer specimens. Ongoing work will illuminate the relationship between endometrial microbiome and reproductive success.

  10. Genetic Polymorphism of the Lactoferrin Gene in Dairy and Beef Cattles at National Artificial Insemination and Embryo Transfer Stations

    Directory of Open Access Journals (Sweden)

    Anneke Anggraeni

    2016-12-01

    Full Text Available Lactoferrin (LTF adalah gen pengontrol komponen protein susu dan memiliki karakteristik sebagai antimikrobial. LTF pada susu berfungsi untuk mencegah diare, sedangkan pada sapi laktasi untuk mencegah mastitis pada ambing. Mempertimbangkan peran penting dari gen LTF, maka perlu dilakukan peningkatan kadar LTF dalam susu melalui seleksi pada taraf DNA. Polymorfisme genetik dari gen LTF diidentifikasi pada sapi perah dan potong dengan metoda Polymerase Chain Reaction - Restricsion Fragment Length Polymorphism (PCR-RFLP, dengan enzim restriksi EcoRI. Genotyping dilakukan pada sapi perah Friesian Holstein (FH total sejumlah 89 ekor, meliputi dari Balai Inseminasi Buatan Lembang (BIB Lembang untuk 17 pejantan, Balai Besar IB Singosari (BBIB Singosari untuk 32 pejantan, dan Balai embrio Transfer Cipelang (BET Cipelang pada 40 dara. Genotyping dilakukan pula pada sapi potong dara berasal dari empat bangsa, meliputi Limousin (14 ekor, Angus (5 ekor, Simmental (13 ekor dan Brahman (5 ekor dari BET Cipelang. Gen LTF|EcoRI pada sapi perah dan potong pengamatan menghasilkan dua tipe alel, yaitu alel A dan B. Kedua jenis sapi tersebut menghasilkan hanya dua genotipe, yaitu genotipe AA dan AB, tanpa genotipe BB. Ini dapat menjadi hal yang baik karena genotipe AA dan AB dipertimbangkan berasosiasi dengan ketahan pada mastitis. Nilai-nilai dari heterozygositas observasi (Ho dari gen ini lebih tinggi dibandingkan heterozigositas ekspektasi (He. Disimpulkan bahwa gen LTF|EcoRI memiliki variasi yang baik pada sapi perah dan sapi potong dari ketiga balai bibit nasional tersebut.

  11. The Chromosomal Constitution of Embryos Arising from Monopronuclear Oocytes in Programmes of Assisted Reproduction

    Directory of Open Access Journals (Sweden)

    Bernd Rosenbusch

    2014-01-01

    Full Text Available The assessment of oocytes showing only one pronucleus during assisted reproduction is associated with uncertainty. A compilation of data on the genetic constitution of different developmental stages shows that affected oocytes are able to develop into haploid, diploid, and mosaic embryos with more or less complex chromosomal compositions. In the majority of cases (~80%, haploidy appears to be caused by gynogenesis, whereas parthenogenesis or androgenesis is less common. Most of the diploid embryos result from a fertilization event involving asynchronous formation of the two pronuclei or pronuclear fusion at a very early stage. Uniparental diploidy may sometimes occur if one pronucleus fails to develop and the other pronucleus already contains a diploid genome or alternatively a haploid genome undergoes endoreduplication. In general, the chance of obtaining a biparental diploid embryo appears higher after conventional in vitro fertilization than after intracytoplasmic sperm injection. If a transfer of embryos obtained from monopronuclear oocytes is envisaged, it should be tried to culture them up to the blastocyst since most haploid embryos are not able to reach this stage. Comprehensive counselling of patients on potential risks is advisable before transfer and a preimplantation genetic diagnosis could be offered if available.

  12. In vitro culture of Arabidopsis embryos.

    Science.gov (United States)

    Sauer, Michael; Friml, Jirí

    2008-01-01

    Embryogenesis of Arabidopsis thaliana follows a nearly invariant cell division pattern and provides an ideal system for studies of early plant development. However, experimental manipulation with embryogenesis is difficult, as the embryo develops deeply inside maternal tissues. Here, we present a method to culture zygotic Arabidopsis embryos in vitro. It enables culturing for prolonged periods of time from the first developmental stages on. The technique omits excision of the embryo by culturing the entire ovule, which facilitates the manual procedure. It allows pharmacological manipulation of embryo development and does not interfere with standard techniques for localizing gene expression and protein localization in the cultivated embryos.

  13. Applications of Tol2 Transposon-Mediated Gene Transfer for Stable Integration and Conditional Expression of Electroporated Genes in Chicken Embryos

    Science.gov (United States)

    Sato, Yuki; Takahashi, Yoshiko

    Because of the high accessibility to developing embryos, avian embryos (chicken and quail) have long been used as a good model animal to study embryogenesis in vertebrates, especially amniotes (reviewed in Wolpert, 2004). The techniques used for “classical” avian embryology included tissue transplantations, tissue ablations, and cell-labeling by vital dye. At the end of the last century, the in ovo electropora tion technique was developed by Nakamura and his colleagues, and this modern method opened a way to study the roles of developmental genes directly in living embryos (Funahashi et al., 1999) reviewed in (Nakamura et al., 2004; Yasuda et al., 2000; Yasugi and Nakamura, 2000). This powerful technique allows us to introduce genes (DNA, RNA, morpholino) into embryos in a tissue-specific way by targeting a restricted area of embryonic tissues. Thus, the electroporation technique using chickens has provided numerous novel insights into the understanding of early development in vertebrates, making the chicken a unique model animal.

  14. Pollination and embryo development in Brassica rapa L. in microgravity

    Science.gov (United States)

    Kuang, A.; Popova, A.; Xiao, Y.; Musgrave, M. E.

    2000-01-01

    Plant reproduction under spaceflight conditions has been problematic in the past. In order to determine what aspect of reproductive development is affected by microgravity, we studied pollination and embryo development in Brassica rapa L. during 16 d in microgravity on the space shuttle (STS-87). Brassica is self-incompatible and requires mechanical transfer of pollen. Short-duration access to microgravity during parabolic flights on the KC-135A aircraft was used initially to confirm that equal numbers of pollen grains could be collected and transferred in the absence of gravity. Brassica was grown in the Plant Growth Facility flight hardware as follows. Three chambers each contained six plants that were 13 d old at launch. As these plants flowered, thin colored tape was used to indicate the date of hand pollination, resulting in silique populations aged 8-15 d postpollination at the end of the 16-d mission. The remaining three chambers contained dry seeds that germinated on orbit to produce 14-d-old plants just beginning to flower at the time of landing. Pollen produced by these plants had comparable viability (93%) with that produced in the 2-d-delayed ground control. Matched-age siliques yielded embryos of equivalent developmental stage in the spaceflight and ground control treatments. Carbohydrate and protein storage reserves in the embryos, assessed by cytochemical localization, were also comparable. In the spaceflight material, growth and development by embryos rescued from siliques 15 d after pollination lagged behind the ground controls by 12 d; however, in the subsequent generation, no differences between the two treatments were found. The results demonstrate that while no stage of reproductive development in Brassica is absolutely dependent upon gravity, lower embryo quality may result following development in microgravity.

  15. Reptile Embryos Lack the Opportunity to Thermoregulate by Moving within the Egg.

    Science.gov (United States)

    Telemeco, Rory S; Gangloff, Eric J; Cordero, Gerardo A; Mitchell, Timothy S; Bodensteiner, Brooke L; Holden, Kaitlyn G; Mitchell, Sarah M; Polich, Rebecca L; Janzen, Fredric J

    2016-07-01

    Historically, egg-bound reptile embryos were thought to passively thermoconform to the nest environment. However, recent observations of thermal taxis by embryos of multiple reptile species have led to the widely discussed hypothesis that embryos behaviorally thermoregulate. Because temperature affects development, such thermoregulation could allow embryos to control their fate far more than historically assumed. We assessed the opportunity for embryos to behaviorally thermoregulate in nature by examining thermal gradients within natural nests and eggs of the common snapping turtle (Chelydra serpentina; which displays embryonic thermal taxis) and by simulating thermal gradients within nests across a range of nest depths, egg sizes, and soil types. We observed little spatial thermal variation within nests, and thermal gradients were poorly transferred to eggs. Furthermore, thermal gradients sufficiently large and constant for behavioral thermoregulation were not predicted to occur in our simulations. Gradients of biologically relevant magnitude have limited global occurrence and reverse direction twice daily when they do exist, which is substantially faster than embryos can shift position within the egg. Our results imply that reptile embryos will rarely, if ever, have the opportunity to behaviorally thermoregulate by moving within the egg. We suggest that embryonic thermal taxis instead represents a play behavior, which may be adaptive or selectively neutral, and results from the mechanisms for behavioral thermoregulation in free-living stages coming online prior to hatching.

  16. Individual demands of human embryos on IVF culture medium: influence on blastocyst development and pregnancy outcome.

    Science.gov (United States)

    Wirleitner, B; Vanderzwalmen, P; Stecher, A; Zech, M H; Zintz, M; Zech, N H

    2010-12-01

    The elucidation of the metabolic requirements of human embryos in vivo or in vitro remains, despite being intensively investigated, a work in progress. The adoption of extended embryo culture to the blastocyst stage during the last decade has entailed new challenges. With the increased attention to culture media formulations, more evidence on the sensitivity of embryos to their early environmental conditions is accumulating which might affect phenotype and developmental potential. A retrospective study was conducted that comprised 286 IVF cycles to evaluate the effect of two different culture media on blastocyst development and pregnancy outcome. Embryos were either cultured in a one step or a sequential medium. Higher fertilization rates and augmented blastocyst rates as well as higher implantation rates were observed when embryos were cultured in one step medium (Pcultured in either medium resulted in a significantly higher rate of twin pregnancies. Although multiple pregnancies should be avoided in assisted reproduction treatment to reduce risks for offspring and mother, this higher frequency of twin pregnancies resulting from the transfer of embryos derived from different culture media suggests that each embryo makes individual demands on its early environment. Copyright © 2010 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  17. Embryo cell allocation patterns are not altered by biopsy but can be linked with further development.

    Science.gov (United States)

    Sepulveda-Rincon, L P; Islam, N; Marsters, P; Campbell, B K; Beaujean, N; Maalouf, W E

    2017-12-01

    It has been suggested that first embryo cleavage can be related with the embryonic-abembryonic axis at blastocyst stage in mice. Thus, cells of the 2-cell embryo might be already biased to form the inner cell mass or trophectoderm. This study was conducted to observe the possible effects of embryo biopsy on cell allocation patterns during embryo preimplantation in two different mouse strains and the effects of these patterns on further development. First, one blastomere of the 2-cell embryo was injected with a lipophilic tracer and cell allocation patterns were observed at blastocyst stage. Blastocysts were classified into orthogonal, deviant or random pattern. For the first experiment, embryos were biopsied at 8-cell stage and total cell counts (TCC) were annotated. Furthermore, non-biopsied blastocysts were transferred into foster mothers. Then, pups and their organs were weighed two weeks after birth. Random pattern was significantly recurrent (≈60%), against orthogonal (patterns among groups. These patterns were not affected by biopsy procedure. However, TCC on deviant embryos were reduced after biopsy. Moreover, no differences were found between patterns for implantation rates, litter size, live offspring and organ weights (lungs, liver, pancreas and spleen). However, deviant pups presented heavier hearts and orthogonal pups presented lighter kidneys among the group. In conclusion, these results suggest that single blastomere removal does not disturb cell allocation patterns during pre-implantation. Nonetheless, the results suggest that embryos following different cell allocation patterns present different coping mechanisms against in vitro manipulations and further development might be altered. © 2017 Society for Reproduction and Fertility.

  18. Efeito do ibuprofeno administrado uma hora antes da inovulação de embriões bovinos Effect of ibuprofen administered one hour before the bovine embryo transfer

    Directory of Open Access Journals (Sweden)

    H.J. Narváez

    2010-06-01

    Full Text Available Avaliou-se o efeito do ibuprofeno administrado uma hora antes da inovulação de embriões bovinos, com o objetivo de melhorar a taxa de prenhez. Após a avaliação da resposta ao protocolo de sincronização do estro, 76 fêmeas selecionadas como receptoras de embriões foram distribuídas em três grupos (G experimentais: G1 (n=25 receptoras usadas como controle, G2 (n=30 receptoras que receberam ibuprofeno 5mg/kg, I.M, uma hora antes da inovulação dos embriões, e G3 (n=21 receptoras que receberam uma matriz polimérica de liberação controlada de ibuprofeno administrado por via subcutânea. As taxas de prenhez foram de 16% (4/25, 43,3% (13/30 e 14,2% (3/21, para G1, G2 e G3, respectivamente. Observou-se diferença (PThe effect of the administered ibuprofen was evaluated one hour before the embryo transfer of bovine embryos in order to improve pregnancy rates. After evaluating the response to protocol synchronization of estrus, 76 Females selected as the recipients of embryos were distributed into three experimental groups: G1 (n = 25 surrogate cows used as control, G2 (n = 30 surrogate cows that received 5mg/kg ibuprofen, IM, one hour before the embryo transfer, and G3 (n = 20 surrogate cows that received an array polymeric release of controlled ibuprofen subcutaneously administered. The pregnancy rates were 16% (4/25, 43.3% (13/30, and 14.2% (3/21 for G1, G2, and G3, respectively. There was statistical difference (P<0.024 on pregnancy rate of G2, in comparison with those of G1 and G3. The administration of ibuprofen intramuscularly one hour before the embryo transfer resulted in better pregnancy rate in Nellore surrogate cows.

  19. Estádios de desenvolvimento embrionário e localização do embrião zigótico em sementes de citros Embryo development stage and the location of embryo zygotic in the seed of citrine

    Directory of Open Access Journals (Sweden)

    Valtemir Gonçalves Ribeiro

    1999-08-01

    Full Text Available Objetivou-se estudar o comportamento de embriões zigóticos e nucelares aos 120, 130, 140 e 150 dias após serem efetuadas hibridações controladas entre a laranjeira 'Natal' (Citrus sinensis Osb. e o parental masculino Poncirus trifoliata (L. Raf. Em cada data, as sementes foram removidas, e os embriões excisados foram caracterizados em estádios de desenvolvimento (globular, cordiforme, torpedo e cotiledonar; coloração (clorofilado ou não; e localização na semente (próximo à micrópila ou mais interiormente. A partir dessas características, foram construídas tabelas de contingência para testar hipóteses de independência entre elas, mediante o teste exato de Fisher e chi² (qui-quadrado. Relações de dependência foram verificadas entre as características: estádios de desenvolvimento embrionário com a localização na semente; estádios de desenvolvimento embrionário com a natureza da plântula (zigótica ou nucelar; e entre natureza da plântula com a localização do embrião na semente. Verificou-se que os embriões zigóticos excisados de frutos com 130 a 150 dias da hibridação controlada, localizam-se, em grande maioria, próximos à região micropilar da semente, em estádio globular e cordiforme de desenvolvimento.Controlled hybridizations between 'Natal' orange variety (Citrus sinensis Osb. and male parent Poncirus trifoliata (L. Raf. were performed in order to study zygotic and nucellar embryos behaviour at 120, 130, 140 and 150 days. At very date the seeds were removed and the embryos excised to characterize: development stage (globular, cordiform, torpedo and cotyledonal; colour (with or without chlorophyll; and location in the seed (close to the micropyle or more internal in the seed. From these characteristics contingency tables were evaluated to test independency hypothesis through Fisher test and chi² (chi-square. Dependence relationships were observed between the following characteristics: stages of the

  20. Methanol as a cryoprotectant for equine embryos.

    Science.gov (United States)

    Bass, L D; Denniston, D J; Maclellan, L J; McCue, P M; Seidel, G E; Squires, E L

    2004-09-15

    Equine embryos (n=43) were recovered nonsurgically 7-8 days after ovulation and randomly assigned to be cryopreserved in one of two cryoprotectants: 48% (15M) methanol (n=22) or 10% (136 M) glycerol (n=21). Embryos (300-1000 microm) were measured at five intervals after exposure to glycerol (0, 2, 5, 10 and 15 min) or methanol (0, 15, 35, 75 and 10 min) to determine changes (%) in diameter over time (+/-S.D.). Embryos were loaded into 0.25-ml plastic straws, sealed, placed in a programmable cell freezer and cooled from room temperature (22 degrees C) to -6 degrees C. Straws were then seeded, held at -6 degrees C for 10 min and then cooled to -33 degrees C before being plunged into liquid nitrogen. Two or three embryos within a treatment group were thawed and assigned to be either cultured for 12 h prior to transfer or immediately nonsurgically transferred to a single mare. Embryo diameter decreased in all embryos upon initial exposure to cryoprotectant. Embryos in methanol shrank and recovered slightly to 76+/-8 % of their original diameter; however, embryos in glycerol continued to shrink, reaching 57+/-6 % of their original diameter prior to cryopreservation. Survival rates of embryos through Day 16 of pregnancy were 38 and 23%, respectively (P>0.05) for embryos cryopreserved in the presence of glycerol or methanol. There was no difference in pregnancy rates of mares receiving embryos that were cultured prior to transfer or not cultured (P>0.05). Preliminary experiments indicated that 48% methanol was not toxic to fresh equine embryos but methanol provided no advantage over glycerol as a cryoprotectant for equine blastocysts.

  1. Occult abnormal pregnancies after first post-embryo transfer serum beta-human chorionic gonadotropin levels of 1.0-5.0 mIU/mL.

    Science.gov (United States)

    Maslow, Bat-Sheva L; Bartolucci, Alison; Sueldo, Carolina; Engmann, Lawrence; Benadiva, Claudio; Nulsen, John C

    2016-04-01

    To assess the occult pregnancy rate after "negative" first post-embryo transfer (ET) serum β-hCG results. Two-part retrospective cohort study and nested case series. University-based fertility center. A total of 1,571 negative first post-ET serum β-hCG results were included in the study; 1,326 results (primary cohort, June 2009-December 2013) were initially reported as <5 mIU/mL and 245 results (secondary cohort, January 2014-March 2015) were reported as discrete values from 1.0 to 5.0 mIU/mL. None. Rates of occult pregnancy, ectopic pregnancy, and complications after negative first post-ET serum β-hCG results. A total of 88.8% (1,178/1,326) of the negative first post-ET results reported as <5 were actually <1.0 mIU/mL. Occult pregnancy was incidentally identified in 1.2% (12/1,041) of subjects with follow-up. Six had ectopic pregnancies, and seven experienced serious complications; 11 (91.7%) of the 12 occult pregnancies had a first post-ET serum β-hCG level of 1.0-5.0 mIU/mL and 1 (8.3%) <1.0 mIU/mL. All pregnancies with serious complications had initial β-hCG levels of 1.0-5.0 mIU/mL. Of the 245 results reported as discreet values, occult pregnancies were diagnosed in 5.5% (9/163) of subjects with follow-up. One had an ectopic pregnancy, which was treated with methotrexate. There were no serious complications in the secondary cohort. The majority of negative first post-ET serum β-hCG levels are <1.0 mIU/mL. Results from 1.0 to 5.0 mIU/mL may fail to exclude abnormal pregnancy and are associated with poor outcomes compared with β-hCG levels <1.0 mIU/mL. Serial serum β-hCG may be warranted in this population. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  2. Diagnosis of human preimplantation embryo viability.

    Science.gov (United States)

    Gardner, David K; Meseguer, Marcos; Rubio, Carmen; Treff, Nathan R

    2015-01-01

    Transfer of more than a single embryo in an IVF cycle comes with the finite possibility of a multiple gestation. Even a twin pregnancy confers significant risk to both mother and babies. The move to single-embryo transfer for all patients will be greatly facilitated by the ability to quantify embryo viability. Developments in time-lapse incubation systems have provided new insights into the developmental kinetics of the human preimplantation embryo. Advances in molecular methods of chromosomal analysis have created platforms for highly effective screening of biopsied embryos, while noninvasive analysis of embryo physiology reveals more about the embryo than can be determined by morphology alone. Recent developments in time-lapse microscopy, molecular karyotyping and in proteomics and metabolomics have been assessed and presented here in a descriptive review. New algorithms are being created for embryo selection based on their developmental kinetics in culture, and the impact of factors such as patient etiology and treatment are being clarified. Potential links between morphokinetic data and embryo karyotype are being elucidated. The introduction of new molecular methods of determining embryo chromosomal complement is proving to be accurate and reproducible, with the future trending toward CGH arrays or next generation sequencing as a rapid and reliable means of analysis, that should be suitable for each IVF clinic to adopt. A relationship between embryo metabolism and viability is established and is now being considered together with morphokinetic data to create more robust algorithms for embryo selection. Microfluidic devices have the capacity and potential to be used in human IVF clinics for the routine diagnosis of embryo biomarkers. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Fresh versus frozen embryo transfer after gonadotropin-releasing hormone agonist trigger in gonadotropin-releasing hormone antagonist cycles among high responder women: A randomized, multi-center study

    Directory of Open Access Journals (Sweden)

    Abbas Aflatoonian

    2018-02-01

    Full Text Available Background: The use of embryo cryopreservation excludes the possible detrimental effects of ovarian stimulation on the endometrium, and higher reproductive outcomes following this policy have been reported. Moreover, gonadotropin-releasing hormone agonist trigger in gonadotropin-releasing hormone (GnRH antagonist cycles as a substitute for standard human chorionic gonadotropin trigger, minimizes the risk of ovarian hyperstimulation syndrome (OHSS in fresh as well as frozen embryo transfer cycles (FET. Objective: To compare the reproductive outcomes and risk of OHSS in fresh vs frozen embryo transfer in high responder patients, undergoing in vitro fertilization triggered with a bolus of GnRH agonist. Materials and Methods: In this randomized, multi-centre study, 121 women undergoing FET and 119 women undergoing fresh ET were investigated as regards clinical pregnancy as the primary outcome and the chemical pregnancy, live birth, OHSS development, and perinatal data as secondary outcomes. Results: There were no significant differences between FET and fresh groups regarding chemical (46.4% vs. 40.2%, p=0.352, clinical (35.8% vs. 38.3%, p=0.699, and ongoing (30.3% vs. 32.7%, p=0.700 pregnancy rates, also live birth (30.3% vs. 29.9%, p=0.953, perinatal outcomes, and OHSS development (35.6% vs. 42.9%, p=0.337. No woman developed severe OHSS and no one required admission to hospital. Conclusion: Our findings suggest that GnRHa trigger followed by fresh transfer with modified luteal phase support in terms of a small human chorionic gonadotropin bolus is a good strategy to secure good live birth rates and a low risk of clinically relevant OHSS development in in vitro fertilization patients at risk of OHSS.

  4. Bovine in-vitro embryo production and its contribution towards ...

    African Journals Online (AJOL)

    Bovine in-vitro embryo production and its contribution towards improved food security in Kenya. ... African Journal of Food, Agriculture, Nutrition and Development ... be low valued, it can be raised to higher reproductive potential using current reproductive technologies such as in-vitro embryo production and embryo transfer.

  5. Ultrastructural changes in goat interspecies and intraspecies reconstructed early embryos

    DEFF Research Database (Denmark)

    Tao, Yong; Gheng, Lizi; Zhang, Meiling

    2008-01-01

    The low efficiency of somatic cell nuclear transfer may be related to the ultrastructural deviations of reconstructed embryos. The present study investigated ultrastructural differences between in vivo-produced and cloned goat embryos, including intra- and interspecies embryos. Goat ear fibroblas...

  6. COMPARAÇÃO ENTRE DOIS MEIOS PARA TRANSFERÊNCIA DE EMBRIÕES EM ÉGUAS DA RAÇA MANGALARGA MARCHADOR OMPARISON BETWEEN TWO MEDIUM FOR EMBRYO TRANSFER IN MARES FROM MANGALARGA MARCHADOR BREED

    Directory of Open Access Journals (Sweden)

    José Renato Costa Caiado

    2009-09-01

    Full Text Available

    Diferentes tampões são utilizados nos meios de lavagem e manutenção embrionária durante os procedimentos de transferência de embriões em equinos (TEE. Os mais usados são carbonato, fosfato e zwitteriônico. Durante os procedimentos de transferência é comum que o embrião permaneça por um tempo maior que o previsto em espera no meio de manutenção utilizado. Este experimento foi realizado com o objetivo de comparar o custo–benefício de dois meios contendo tampões diferentes DPBS (fosfato e Embriocare® (zwitteriônico, na transferência de embriões em éguas da raça Mangalarga Marchador. Dividiram-se os embriões aleatoriamente em dois tratamentos – zwitteriônico e fosfato –, permanecendo em descanso por 0, 30, 60 ou 120 minutos em ambos os meios. Os resultados indicam que esses tampões são igualmente eficientes (P > 0,05 nos processos de rasteamento, lavagem, manutenção à temperatura ambiente por diferentes períodos de tempo (0 a 120 minutos, sendo que as taxas de prenhez obtidas em ambiente tropical foram de 65% e de 69,7%, respectivamente. Porém, obteve-se a maior taxa de prenhez encontrada com DPBS com um tempo de manutenção do embrião por sessenta minutos no meio (P < 0,05. No tampão zwitteriônico, a maior taxa de prenhez foi alcançada com trinta minutos no meio (P < 0,05. Como a aquisição do Embriocare® foi mais onerosa, o DPBS apresentou o melhor custo–benefício.

    PALAVRAS-CHAVES: Equino, Mangalarga Marchador, transferência de embriões, tampão fosfato e zwitteriônico.

    Palavras-chave: eqüino, Mangalarga Marchador, transferência de embriões, tampão fosfato e zwitteriõnico. Different buffers are used in the solution for washing and embryo maintenance during the procedures of equine embryo transfer

  7. Which factors are most predictive for live birth after in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) treatments? Analysis of 100 prospectively recorded variables in 8,400 IVF/ICSI single-embryo transfers.

    Science.gov (United States)

    Vaegter, Katarina Kebbon; Lakic, Tatevik Ghukasyan; Olovsson, Matts; Berglund, Lars; Brodin, Thomas; Holte, Jan

    2017-03-01

    To construct a prediction model for live birth after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment and single-embryo transfer (SET) after 2 days of embryo culture. Prospective observational cohort study. University-affiliated private infertility center. SET in 8,451 IVF/ICSI treatments in 5,699 unselected consecutive couples during 1999-2014. A total of 100 basal patient characteristics and treatment data were analyzed for associations with live birth after IVF/ICSI (adjusted for repeated treatments) and subsequently combined for prediction model construction. Live birth rate (LBR) and performance of live birth prediction model. Embryo score, treatment history, ovarian sensitivity index (OSI; number of oocytes/total dose of FSH administered), female age, infertility cause, endometrial thickness, and female height were all independent predictors of live birth. A prediction model (training data set; n = 5,722) based on these variables showed moderate discrimination, but predicted LBR with high accuracy in subgroups of patients, with LBR estimates ranging from 40%. Outcomes were similar in an internal validation data set (n = 2,460). Based on 100 variables prospectively recorded during a 15-year period, a model for live birth prediction after strict SET was constructed and showed excellent calibration in internal validation. For the first time, female height qualified as a predictor of live birth after IVF/ICSI. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. Transcervical Embryo recovery by transcervical technique in ...

    African Journals Online (AJOL)

    Vicente Freitas

    1970s that similar technologies were adopted in goat breeding. Since then, embryos from thousands of goats have been collected and transferred. Three countries, Australia, New Zealand and South Africa, are particularly prominent in this field ...

  9. ANÁLISE DA PRODUÇÃO DE EMBRIÕES NA FERTILIZAÇÃO IN VITRO E TRANSFERÊNCIA DE EMBRIÕES PARA DOADORAS NELORE ANALYSIS OF EMBRYO PRODUCTION IN VITRO FERTILIZATION AND EMBRYO TRANSFER TO NELLORE DONORS

    Directory of Open Access Journals (Sweden)

    Renato Travassos Beltrame

    2010-04-01

    Full Text Available

    Ajustou-se uma função de densidade probabilidade para o
    número de embriões viáveis produzidos após fertilização in vitro
    em doadoras da raça Nelore, a partir de dados fornecidos pela Associação Brasileira de Criadores de Zebu (ABCZ, referente à análise de 20.619 doadoras, 71.602 aspirações e um total de 509.643 embriões. Modelou-se a densidade probabilidade do número de embriões viáveis mediante a função exponencial, executando-se a determinação dos parâmetros por meio da máxima verossimilhança, em um método de gradiente não linear. O nível de precisão obtido foi de RMSE = 0,040 e R2 = 0,98, para a representação da probabilidade do número de embriões viáveis produzidos por doadoras Nelore na técnica de fertilização in vitro(FIV. Para comparar os modelos (curvas de probabilidade de transferência de embriões ajustada por Beltrame, em 2006, e de FIV, neste trabalho, aplicou-se a técnica de comparação de curvas com o teste F (Silva e Azevedo, 2002. Não foram encontradas diferenças entre as curvas do número de embriões viáveis obtidos após coleta e produzidos após aspiração de doadoras na raça Nelore. Ainda, sugere-se a existência de um fator único limitante que afete biologicamente a produção de embriões nas técnicas de transferência de embriões e fertilização in vitro.

    PALAVRAS-CHAVES: Banco de dados, densidade probabilidade, doadoras, simulação.

    Aprobability density function for the number of viable embryos produced after an in vitro fertilization program in Nellore donors  was adjusted through data provided by the Brazilian Association of Zebu breeders. Results were based on 20,619 donors, 71,602 aspirations and the total of 509,643 embryos. The probability density function of the number of viable embryos was modeled using exponential distribution. Parameters fitting were carried out for the maximum likelihood using a non-linear gradient method. The

  10. Desempenho de diferentes estádios embrionários no cultivo in vitro de embriões de 'Pêra Rio' x 'Poncã' Perfomance of different embryo stage of sweet orange x mandarin cultivated in vitro

    Directory of Open Access Journals (Sweden)

    Edvan Alves Chagas

    2003-12-01

    Full Text Available Objetivou-se verificar qual o melhor estádio embrionário para o cultivo de embriões imaturos oriundos de frutos provenientes de hibridação entre 'Pêra Rio' x 'Poncã' , bem como o efeito de diferentes concentrações do meio de cultura MT. Os embriões em diferentes estádios de desenvolvimento (globulares, torpedo e cordiforme foram excisados e inoculados em tubos de ensaio contendo 15 mL do meio MT com diferentes concentrações (0; 50; 100 e 150% da composição original e acrescido de 50 g.L-1 de sacarose. Após a inoculação, os embriões foram incubados à 27±1ºC, fotoperíodo de 16 horas e irradiância de 32 mmol.m-2.s-1. Após 90 dias, avaliou-se o comprimento da parte aérea e do sistema radicular, massa fresca e número de folhas das plântulas. Melhor desenvolvimento dos embriões imaturos foi obtido em estádio cotiledonar e com a concentração de 150% do meio MT.Embryos obtained from Citrus sinensis Osbeck 'Pêra Rio' x Citrus reticulata Blanco 'Poncã' crossings were excised in diverse phases of development and cultivated in different medium concentrations. Embryos in globular, torped and cotiledonary stage were excised and inoculated individually in test tubes with 15 mL in distinct MT medium concentrations (0; 50; 100 and 150% of original composition and added sucrose (50 g.L-1. After inoculation, the embryos were maintained in growth room at 27±1ºC, 16 hour photoperiod and 32 umol.m-2.s-1 irradiance. After 90 days, the length of the aerial part and roots system, fresh mass and number of leaves, were evaluated. The best results were obtained by incubating embryos in the cotiledonary stage, in MT medium with concentrations of 150%.

  11. Cryopreservation of embryos and oocytes in human assisted reproduction.

    Science.gov (United States)

    Konc, János; Kanyó, Katalin; Kriston, Rita; Somoskői, Bence; Cseh, Sándor

    2014-01-01

    Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification) of human embryos and oocytes are summarized.

  12. Cryopreservation of Embryos and Oocytes in Human Assisted Reproduction

    Directory of Open Access Journals (Sweden)

    János Konc

    2014-01-01

    Full Text Available Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification of human embryos and oocytes are summarized.

  13. Dynamics of progesterone, TNF-a and a metabolite of PGF2a in blood plasma of beef cows following embryo transfer

    Science.gov (United States)

    Lactating beef cows received an embryo along with no treatment (control; n = 16), controlled internal drug releasing device (CIDR; n = 16), human chorionic gonadotropin (hCG; n = 15), or gonadotropin releasing hormone (GnRH; n = 15) to assess the effectiveness of these treatments in increasing blood...

  14. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-12-01

    Full Text Available http://dx.doi.org/10.5902/1980509812343Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germinationand that makes its natural propagation difficult. The aim of this study was to establish a protocol ofregeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculatedon WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein orglutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D(22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed caseinor glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture mediumcontaining NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction(8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein andthe development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promotedin WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containinghydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. Duringthe maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages.The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histologicalstudies.

  15. SOMATIC EMBRYOGENESIS AND MORPHOANATOMY OF Ocotea porosa SOMATIC EMBRYOS

    Directory of Open Access Journals (Sweden)

    Luciana Luiza Pelegrini

    2013-01-01

    Full Text Available Ocotea porosa seeds have strong tegument dormancy, recalcitrant behavior, low and irregular germination and that makes its natural propagation difficult. The aim of this study was to establish a protocol of regeneration of Ocotea porosa from somatic embryogenesis. Immature embryonic axes were inoculated on WPM culture medium supplemented with 2.4-D (200 μM combined or not with hydrolyzed casein or glutamine (0.5 or 1 g l-1, during 90 days. The repetitive embryogenesis was induced on medium with 2.4-D (22.62 μM combined with 2-iP (2.46 μM followed by transfer to culture medium with hydrolyzed casein or glutamine (1 g l-1 during 90 days. The maturation of somatic embryos was tested in culture medium containing NAA (0.5 μM and 2-iP (5; 10 and 20 μM. The highest percentage of somatic embryos induction (8.3% was observed in WPM culture medium containing 200 μM 2.4-D and 1 g L-1 hydrolyzed casein and the development of somatic embryos occurred indirectly. Repetitive somatic embryogenesis was promoted in WPM medium containing hydrolyzed casein or glutamine. However, the culture medium containing hydrolyzed casein promoted the maintenance of embryogenic capacity for more than two years. During the maturity phase, there was a low progression of globular embryos to cordiform and torpedo stages. The different ontogenetic stages of somatic embryos of Ocotea porosa were characterized by histological studies.

  16. The progress in diagnostic imaging for staging of bladder and prostate cancer. Endorectal magnetic resonance imaging and magnetization transfer contrast

    Energy Technology Data Exchange (ETDEWEB)

    Arima, Kiminobu; Hayashi, Norio; Yanagawa, Makoto; Kawamura, Juichi; Kobayashi, Shigeki; Takeda, Kan [Mie Univ., Tsu (Japan). School of Medicine; Sugimura, Yoshiki

    1999-08-01

    We retrospectively studied the staging accuracy of endorectal magnetic resonance imaging (MRI) in comparison with transrectal ultrasound examination (TRUS) for 71 localized bladder cancers and 19 localized prostate cancers (PC) radically resected. The accuracy of clinical staging for bladder cancer in endorectal MRI and TRUS was 85.9% and 69.2%, respectively. The presence or absence of the continuity of submucosal enhancement on T2-weighted MRI images could be useful for the staging of bladder cancer. The accuracy of the seminal vesicular invasion for prostate cancer in endorectal MRI and TRUS was 95% and 63%, respectively. To determine whether magnetization transfer contrast (MTC) provides additional information in the diagnosis of prostate cancer, the magnetization transfer ratios (MTRs) were calculated in 22 patients with PC, 5 with benign prostatic hyperplasia (BPH) and 4 controls. The mean MTR in the peripheral zone of the normal prostate (8.0%{+-}3.4 [standard deviation]) showed a statistically significant decrease relative to that in the inner zone of the normal prostate (27.4%{+-}3.4, p<0.01), BPH (25.5%{+-}3.7, p<0.01), pre-treatment PC (30.6%{+-}5.9, p<0.01), and PC after hormonal therapy (20.3%{+-}6.3, p<0.01). The mean MTR in pre-treatment PC was significantly higher than that in BPH, or in PC after hormonal therapy (p<0.01). MTC was considered to be useful for conspicuity of prostate cancer lesion. (author)

  17. Mouse embryos cultured in amniotic fluid | Oettle | South African ...

    African Journals Online (AJOL)

    The embryos were cultured for 72 hours at 37°C in 5% carbon dioxide in air. Osmolarity, pH, partial arterial carbon dioxide pressure and HCO; were checked before culture and again at the end of culture. Embryos were assessed according to the stage of development, namely degenerate embryos, morulae and blastocysts.

  18. Xenopus laevis embryos and tadpoles as models for testing for ...

    African Journals Online (AJOL)

    The toxicity of bio available Zn, Cu, Pb, and Cd on the life stages of Xenopus laevis embryos and tadpoles was investigated. Cu and Cd were found to affect the hatching success of the embryos, with a strong negative relationship existing between the increase in Cu concentrations and the hatching of the embryos.

  19. Superovulation Response and In vivo Embryo Production Potential ...

    African Journals Online (AJOL)

    to a search dish containing holding medium and grouped in to embryos and UFO under a stereomicroscope (MotisSMZ 140/143®, Roanoke, USA). The embryos were graded for their developmental stage (from stage 1 = 1-cell to stage 9 = expanded hatched blastocyst) and quality (from quality 1 = excellent to quality 4.

  20. Post-implantation mortality of in vitro produced embryos is associated with DNA methyltransferase 1 dysfunction in sheep placenta.

    Science.gov (United States)

    Ptak, Grazyna Ewa; D'Agostino, Antonella; Toschi, Paola; Fidanza, Antonella; Zacchini, Federica; Czernik, Marta; Monaco, Federica; Loi, Pasqualino

    2013-02-01

    Is DNA methyltransferase 1 (DNMT1) dysfunction involved in epigenetic deregulation of placentae from embryos obtained by assisted reproduction technologies (ARTs)? DNMT1 expression in growing placentae of in vitro produced (IVP) embryos is compromised and associated with pregnancy loss. DNMT1 maintains the methylation profile of genes during cell division. The methylation status of genes involved in placenta development is altered in embryos obtained in vitro. Disturbances in the epigenetic regulation of gene expression during placentogenesis could be involved in the frequent developmental arrest and loss of IVP embryos. Forty sheep were naturally mated (Group 1, CTR). IVP blastocysts (2-4 per ewe) were surgically transferred to the remaining 46 recipient sheep 6 days after oestrus (Group 2). Twenty-one recipients from Group 1 and 27 recipients from Group 2 were allowed to deliver in order to compare embryo survival in both groups at term (150 days). From the remaining recipients (n = 38), fetuses and placentae of both groups were recovered by paramedian laparotomy at Days 20, 22, 24, 26 and 28 of gestation. Immediately after collection, early placental tissues (chorion-allantois) were snap frozen in liquid nitrogen and DNMT1 expression and activity was evaluated. mRNA levels (for DNMT1, HDAC2, PCNA, DMAP1, MEST, IGF2, CDKN1C, H19) and the methylation status of H19 were also analyzed. Furthermore, embryo size and survival rate were measured. Our study shows that DNMT1 expression was reduced in early placentae from sheep IVP embryos. This reduction was associated with growth arrest and subsequent death of the sheep embryos. Conversely, normal levels of DNMT1 and its cofactors were observed in placentae from IVP embryos that survived this developmental bottleneck. Although DNA methylation machinery was severely compromised in IVP placentae only up to Day 24, the low DNMT1 enzymatic activity that persisted after this stage in IVP placentae was not lethal for the

  1. Developmental competence of porcine chimeric embryos produced by aggregation

    DEFF Research Database (Denmark)

    Li, Juan; Jakobsen, Jannik E.; Xiong, Qiang

    2015-01-01

    either by parthenogenetic activation (PA) or handmade cloning (HMC). Results showed that the developmental competence of chimeric embryos, evaluated based on their blastocyst rate and total cell number per blastocyst, was increased when two whole 2-cell stage embryos (PA or HMC) were aggregated....... In comparison, when two blastomeres were aggregated, the developmental competence of the chimeric embryos decreased if the blastomeres were either from PA or from HMC embryos, but not if they were from different sources, i.e. one PA and one HMC blastomere. To evaluate the cell contribution in embryo formation......, aggregation was made with HMC embryos cloned using EGFP transgenic cells; the cell contribution in the formation of the inner cell mass or trophectoderm was random in chimeric blastocysts. Finally, two blastomeres from 2-cell stage embryos were fused to construct tetraploid embryos, and when diploid...

  2. Post-hatching development of the porcine and bovine embryo-defining criteria for expected development in vivo and in vitro

    DEFF Research Database (Denmark)

    Vejlsted, Morten; Du, Yutao; Vajta, Gábor

    2006-01-01

    without the need for transfer to recipient animals. Such a system would require (1) definition of milestones of expected post-hatching embryonic development in vivo; and (2) development of adequate culture systems. We propose a stereomicroscopical staging system for post-hatching embryos defining...

  3. Cloning Endangered Felids by Interspecies Somatic Cell Nuclear Transfer.

    Science.gov (United States)

    Gómez, Martha C; Pope, C Earle

    2015-01-01

    In 2003, the first wild felid was produced by interspecies somatic cell nuclear transfer. Since then other wild felid clone offspring have been produced by using the same technique with minor modifications. This chapter describes detailed protocols used in our laboratory for (1) the isolation, culture, and preparation of fibroblast cells as donor nucleus, and (2) embryo reconstruction with domestic cat enucleated oocytes to produce cloned embryos that develop to the blastocyst stage in vitro and, after transfer into synchronized recipients, establish successful pregnancies.

  4. Risk of ectopic pregnancy lowest with transfer of single frozen blastocyst.

    Science.gov (United States)

    Li, Z; Sullivan, E A; Chapman, M; Farquhar, C; Wang, Y A

    2015-09-01

    What type of transferred embryo is associated with a lower rate of ectopic pregnancy? The lowest risk of ectopic pregnancy was associated with the transfer of blastocyst, frozen and single