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Sample records for vitro fertilized porcine

  1. Osteopontin reduces polyspermy during in vitro fertilization of porcine oocytes.

    Science.gov (United States)

    Hao, Yanhong; Mathialagan, Nagappan; Walters, Eric; Mao, Jiude; Lai, Liangxue; Becker, Donald; Li, Wensheng; Critser, John; Prather, Randall S

    2006-11-01

    This study was designed to determine the role of osteopontin (SPP1) in in vitro fertilization (IVF) in swine. The initial objective was to evaluate the effect of various concentrations of SPP1 (0, 0.001, 0.01, 0.1 and 1 microg/ml) on spermatozoa and oocytes during IVF. The results demonstrate that SPP1 reduced the rate of polyspermy in a dose-dependent manner (P polyspermy was specific to SPP1, a mixture of pregnancy-associated glycoproteins was included in the IVF protocol and shown to have no effect on polyspermy. Furthermore, Western blotting demonstrated that a 50-kDa SPP1 form was present in the oviducts on Days 0, 3, and 5 in pregnant and nonpregnant gilts, and the concentration of SPP1 on Day 0 was higher than on Days 3 and 5. The current study represents the first report to demonstrate that SPP1 plays an important role in the regulation of pig polyspermic fertilization; it decreases polyspermy and increases fertilization efficiency during IVF.

  2. Effect of Dipeptides on In vitro Maturation, Fertilization and Subsequent Embryonic Development of Porcine Oocytes

    Science.gov (United States)

    Tareq, K. M. A.; Akter, Quzi Sharmin; Tsujii, Hirotada; Khandoker, M. A. M. Yahia; Choi, Inho

    2013-01-01

    The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM)-199 supplemented with 10% (v/v) porcine follicular fluid, modified Tyrode’s albumin lactate pyruvate (mTALP) medium, and modified North Carolina State University (mNCSU)-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln)+L-glycyl-L-glutamine (GlyGln) significantly (p<0.05) improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U)-glucose when compared to the control group and other treatment groups. Additionally, 2–4 cell, 8–16 cell, morula and blastocyst development increased significantly (p<0.05) following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media. PMID:25049815

  3. Improving porcine in vitro fertilization output by simulating the oviductal environment

    Science.gov (United States)

    Soriano-Úbeda, Cristina; García-Vázquez, Francisco A.; Romero-Aguirregomezcorta, Jon; Matás, Carmen

    2017-01-01

    Differences between the in vitro and in vivo environment in which fertilization occurs seem to play a key role in the low efficiency of porcine in vitro fertilization (IVF). This work proposes an IVF system based on the in vivo oviductal periovulatory environment. The combined use of an IVF medium at the pH found in the oviduct in the periovulatory stage (pHe 8.0), a mixture of oviductal components (cumulus-oocyte complex secretions, follicular fluid and oviductal periovulatory fluid, OFCM) and a device that interposes a physical barrier between gametes (an inverted screw cap of a Falcon tube, S) was compared with the classical system at pHe 7.4, in a 4-well multidish (W) lacking oviduct biological components. The results showed that the new IVF system reduced polyspermy and increased the final efficiency by more than 48%. This higher efficiency seems to be a direct consequence of a reduced sperm motility and lower capacitating status and it could be related to the action of OFCM components over gametes and to the increase in the sperm intracellular pH (pHi) caused by the higher pHe used. In conclusion, a medium at pH 8.0 supplemented with OFCM reduces polyspermy and improves porcine IVF output.

  4. A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes.

    Science.gov (United States)

    Park, Chi-Hun; Lee, Sang-Goo; Choi, Don-Ho; Lee, Chang-Kyu

    2009-10-01

    The general method of porcine in vitro fertilization (IVF), involving the co-culture of both gametes in a medium drop, is thought to be the main reason for the high incidence of polyspermy. The aim of this study was to reduce the polyspermic fertilization of porcine embryos during IVF by the modified swim-up method, based on general sperm swim-up technique. Within this design, a 70 microm pore sized cell strainer was used to separate the sperm pellet placed at the bottom of a tube from the mature oocytes placed within the upper region. The separation of gametes using this permeable barrier was to ensure that only motile sperm gained access to the oocytes. It was found that the rate of polyspermy was significantly lowered for the sperm preparations from three boar breeds in modified swim-up method when compared with that of the general microdrop method (p<0.05). However, the penetration rates were found to be similar in both methods for two boar breeds. The average occurrence of blastocysts with more total cell number was higher in the modified swim-up method, while no significant difference in blastocyst rates between the two IVF methods was observed. The frequency of normal diploid embryos was also significantly higher in the modified swim-up method and polyploidy was more frequently observed in microdrop method (p<0.05). Our results demonstrated that the modified swim-up IVF method could reduce polyspermic penetration, and consequently produce better quality and karyotypically normal embryos in porcine IVF.

  5. Effects of the porcine oviduct-specific glycoprotein on fertilization, polyspermy, and embryonic development in vitro.

    Science.gov (United States)

    Kouba, A J; Abeydeera, L R; Alvarez, I M; Day, B N; Buhi, W C

    2000-07-01

    This study evaluated the effects of porcine oviduct-specific glycoprotein (pOSP) on in vitro fertilization (IVF), polyspermy, and development to blastocyst. Experiment 1 evaluated the effects of various concentrations (0-100 microgram/ml) of purified pOSP on fertilization parameters, including penetration, polyspermy, male pronuclear formation, and mean number of sperm penetrated per oocyte. Experiment 2 examined the ability of an anti-pOSP immunoglobulin G to inhibit the observed effects of pOSP on fertilization parameters. Experiments 3 and 4 examined various concentrations of pOSP (0-100 microgram/ml) on zona pellucida solubility and sperm binding, respectively. Lastly, experiment 5 assessed the effects of various concentrations of pOSP (0-100 microgram/ml) on the in vitro embryo cleavage rate and development to blastocyst. Pig oocytes matured and fertilized in vitro were used for all experiments. An effect of treatment (P polyspermy, and mean number of sperm per oocyte. Concentrations for pOSP of 0-50 microgram/ml had no effect on sperm penetration rates; however, compared with the control, 100 microgram/ml significantly decreased the penetration rate (74% vs. 41%). Addition of 10-100 microgram/ml significantly reduced the polyspermy rate compared with the control (61% vs. 24-29%). The decrease in polyspermy achieved by addition of pOSP during preincubation and IVF was blocked with a specific antibody to pOSP. No effect of treatment was observed on zona digestion time relative to the control; however, the number of sperm bound to the zona pellucida was significantly decreased by treatment (P polyspermy in pig oocytes, reduces the number of bound sperm, and increases postcleavage development to blastocyst.

  6. In vitro fertilization of porcine oocytes is affected by spermatic coincubation time

    Directory of Open Access Journals (Sweden)

    Guilherme Oberlender

    Full Text Available Abstract: The aim was to study the effects of different gamete coincubation times on porcine in vitro fertilization (IVF, and to verify whether efficiency could be improved by reducing oocyte exposure time to spermatozoa during IVF. In groups of 50, a total of 508 immature cumulus-oocyte complexes (COCs were matured in NCSU-37 medium. The COCs were cultured for 44 hours and then inseminated with in natura semen (2,000 spermatozoa/oocyte. The sperm and oocytes were coincubated according to the following treatments (T: T1 = oocytes exposed to spermatozoa for one hour (173 oocytes, T2 = oocytes exposed to spermatozoa for two hours (170 oocytes, and T3 = oocytes exposed to spermatozoa for three hours (165 oocytes. After these coincubation periods, the oocytes were washed in fertilization medium (TALP medium to remove spermatozoa not bound to the zona pellucida and cultured in another similar medium (containing no sperm. Eighteen to twenty hours after fertilization, the putative zygotes were stained in Hoechst-33342 to evaluate the IVF results. The penetration rate was higher (P0.05 between oocytes exposed to spermatozoa for one (T1 and three hours (T3. However, optimum (P=0.048 results were obtained after two hours of coincubation, when the rate of fertilization performance was 50.16±8.52%. The number of penetrated sperm per oocyte, as well as male pronucleus formation, did not differ (P>0.05 between the treatments evaluated. Under these assay conditions, especially in relation to the sperm concentration used, gamete coincubation for a period of two hours appears to be optimal for monospermy and fertilization performance. Thus, it is the optimal time period for obtaining a large number of pig embryos capable of normal development.

  7. Inhibition of 19S proteasomal regulatory complex subunit PSMD8 increases polyspermy during porcine fertilization in vitro.

    Science.gov (United States)

    Yi, Young-Joo; Manandhar, Gaurishankar; Sutovsky, Miriam; Jonáková, Vera; Park, Chang-Sik; Sutovsky, Peter

    2010-03-01

    The 26S proteoasome is a multi-subunit protease specific to ubiquitinated substrate proteins. It is composed of a 20S proteasomal core with substrate degradation activity, and a 19S regulatory complex that acts in substrate recognition, deubiquitination, priming and transport to the 20S core. Inhibition of proteolytic activities associated with the sperm acrosome-borne 20S core prevents fertilization in mammals, ascidians and echinoderms. Less is known about the function of the proteasomal 19S complex during fertilization. The present study examined the role of PSMD8, an essential non-ATPase subunit of the 19S complex, in sperm-ZP penetration during porcine fertilization in vitro (IVF). Immunofluorescence localized PSMD8 to the outer acrosomal membrane, acrosomal matrix and the inner acrosomal membrane. Colloidal gold transmission electron microscopy detected PSMD8 on the surface of vesicles in the acrosomal shroud, formed as a result of zona pellucida-induced acrosomal exocytosis. Contrary to the inhibition of fertilization by blocking of the 20S core activities, fertilization and polyspermy rates were increased by adding anti-PSMD8 antibody to fertilization medium. This observation is consistent with a possible role of PSMD8 in substrate deubiquitination, a process which when blocked, may actually accelerate substrate proteolysis by the 26S proteasome. Subunit PSMD8 co-immunoprecipitated with acrosomal surface-associated spermadhesin AQN1. This association indicates that the sperm acrosome-borne proteasomes become exposed onto the sperm surface following the acrosomal exocytosis. Since immunological blocking of subunit PSMD8 increases the rate of polyspermy during porcine fertilization, the activity of the 19S complex may be a rate-limiting factor contributing to anti-polyspermy defense during porcine fertilization. Copyright 2009. Published by Elsevier Ireland Ltd.

  8. Carboxyethylgermanium sesquioxide (Ge-132) treatment during in vitro culture protects fertilized porcine embryos against oxidative stress induced apoptosis.

    Science.gov (United States)

    Kim, Eunhye; Hwang, Seon-Ung; Yoon, Junchul David; Jeung, Eui-Bae; Lee, Eunsong; Kim, Dae Young; Hyun, Sang-Hwan

    2017-12-15

    Compared with the in vivo environment, porcine in vitro embryo-culture systems are suboptimal, as they induce oxidative stress via the accumulation of reactive oxygen species (ROS). High ROS levels during early embryonic development cause negative effects, such as apoptosis. In this study, we examined the effects of the antioxidant carboxyethylgermanium sesquioxide (Ge-132) during in vitro culture (IVC) on embryonic development in porcine in vitro fertilization (IVF) embryos. Zygotes were treated with different concentrations of Ge-132 (0, 100, 200 and 400 μg/ml). All of the Ge-132 treatment groups displayed greater total cell numbers after IVC (98.1, 98.5 and 103.4, respectively) compared with the control group (73.9). The 200 μg/ml Ge-132 treatment group exhibited significantly increased intracellular GSH levels compared with the control group, whereas the ROS generation levels decreased in Ge-132 dose-dependent manner (P cultured under Ge-132 treatment may be associated with KEAP1 signaling cascades involved in oxidative stress and apoptosis during porcine preimplantation embryo development.

  9. Anti-hyaluronidase oligosaccharide derived from chondroitin sulfate a effectively reduces polyspermy during in vitro fertilization of porcine oocytes.

    Science.gov (United States)

    Tatemoto, Hideki; Muto, Norio; Yim, Sun-Deok; Nakada, Tadashi

    2005-01-01

    The present study was conducted to examine the effects of chondroitin sulfate A-derived oligosaccharide (ChSAO) on hyaluronidase activity and in vitro fertilization (IVF) parameters. The activity of hyaluronidase extracted from preincubated boar sperm was completely blocked by ChSAO at concentrations of 10 microg/ml or higher. After in vitro maturation of porcine cumulus-oocyte complexes, some oocytes were freed from their cumulus cells, and cumulus-intact or cumulus-free oocytes were inseminated with sperm in IVF medium containing various concentrations of ChSAO (0.1-100 microg/ml). In cumulus-intact oocytes, the penetration and the polyspermy rates (39% and 28%, respectively) were significantly decreased by treatment with 100 microg/ml ChSAO compared with those of oocytes treated without ChSAO (63% and 52%, respectively). On the contrary, in cumulus-free oocytes, the addition of 10-100 microg/ml ChSAO significantly reduced the polyspermy rate compared with the control (25-30% versus 53%, respectively), whereas ChSAO had no effect on sperm penetration. Interestingly, ChSAO added to IVF medium significantly decreased the number of sperm bound to the zona pellucida (ZP) of cumulus-free oocytes in a concentration-dependent manner between 0.1 and 100 microg/ml. However, ChSAO had no effect on the time course change in ZP modification after oocyte activation by electrostimulation and the incidence of the acrosome-reacted sperm. Treatment with 100 microg/ml ChSAO during IVF of cumulus-free oocytes significantly increased the proportion of development to the blastocyst stage after in vitro insemination. Therefore, the present findings indicate that hyaluronidase-inhibitory ChSAO is an efficient probe for promoting normal fertilization process in terms of an effective decrease in the incidence of polyspermy during IVF of porcine oocytes.

  10. Inhibition of boar sperm hyaluronidase activity by tannic acid reduces polyspermy during in vitro fertilization of porcine oocytes.

    Science.gov (United States)

    Tatemoto, Hideki; Tokeshi, Isao; Nakamura, Satoshi; Muto, Norio; Nakada, Tadashi

    2006-11-01

    The present study was conducted to examine the effects of three polyphenols (tannic acid, apigenin and quercetin) on hyaluronidase activity and in vitro fertilization (IVF) parameters. Among them, tannic acid showed by far the strongest potency for blocking hyaluronidase activity extracted from preincubated boar sperm, causing a dose-dependent inhibition over the range of 2-10 microg/ml. When cumulus-intact and cumulus-free oocytes were inseminated in IVF medium containing tannic acid, the penetration and the polyspermy rates were significantly decreased in the presence of 10 microg/ml tannic acid compared with those in the absence of tannic acid, and the addition of 5 microg/ml tannic acid significantly reduced the polyspermy rate (p polyspermy. Interestingly, the incidence of polyspermy was significantly reduced in oocytes inseminated with sperm pretreated with 5 microg/ml tannic acid (p polyspermy after insemination with untreated sperm. Treatment with tannic acid caused neither a protective proteolytic modification of the zona pellucida matrix before fertilization, nor a reduction of the proteolytic activity of acrosomal contents or the number of zona-bound spermatozoa. These data suggest that an appropriate concentration of tannic acid prevents polyspermy through the inhibition of sperm hyaluronidase activity during IVF of porcine oocytes.

  11. Toxicity evaluation of ethanol treatment during in vitro maturation of porcine oocytes and subsequent embryonic development following parthenogenetic activation and in vitro fertilization.

    Science.gov (United States)

    Lee, Sanghoon; Kim, Eunhye; Hyun, Sang-Hwan

    2014-11-01

    Ethanol is frequently used as a solvent in several techniques for in vitro production (IVP). It is also used for the parthenogenetic activation (PA) of oocytes. Although a number of studies have suggested that ethanol has detrimental effects on fibroblasts and neuronal cells, little attention has been paid to the effects of ethanol on porcine oocytes. Thus, the aim of this study was to evaluate the effects of the addition of ethanol to in vitro maturation (IVM) medium. We investigated the effects of ethanol (0, 1 and 3%) on the following parameters: nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, and subsequent embryonic development following PA and in vitro fertilization (IVF). After 44 h of IVM, the 3% group showed a significant (Pethanol group showed significantly (Pethanol group had significantly (P1% ethanol during IVM exerts a toxic effect on the developmental potential of PA and IVF porcine embryos by decreasing the intracellular GSH level, thereby increasing the intracellular ROS level and upregulating the expression of apoptosis‑related genes.

  12. Cryotolerance of porcine blastocysts is improved by treating in vitro matured oocytes with L-carnitine prior to fertilization

    Science.gov (United States)

    LOWE, Jenna L.; BARTOLAC, Louise K.; BATHGATE, Roslyn; GRUPEN, Christopher G.

    2017-01-01

    Sufficient generation of adenosine triphosphate (ATP) by oocytes is critical for fertilization and embryo development. The objective of this study was to determine the effects of supplementing media with L-carnitine, a co-factor required for the metabolism of fatty acids, during the peri-fertilization period on embryo development and energy generation. Firstly, in vitro matured (IVM) porcine oocytes were co-incubated with sperm in IVF medium supplemented with 0‒24 mM L-carnitine. The blastocyst formation rate of the control group was greater than those of the L-carnitine groups (P L-carnitine group. Subsequently, oocytes and/or sperm were treated without or with 3 mM L-carnitine for either the 1 h pre-IVF oocyte incubation; the pre-IVF sperm preparation; the first 30 min of IVF; or the entire 5.5 h of IVF. Despite similar fertilization rates among the groups, the cleavage rate of the pre-IVF oocyte group was significantly greater than those of the other groups, except for the pre-IVF sperm group. Additionally, the oocyte ATP content and the cryotolerance of the resulting blastocysts were examined following the pre-IVF oocyte treatment. Oocyte ATP content was also similar among the groups (P > 0.05). Following vitrification, the post-warming survival rate of blastocysts derived from L-carnitine-treated oocytes was greater than that of blastocysts derived from untreated oocytes (42.4% vs. 24.9%; P L-carnitine immediately prior to insemination enhanced cleavage and improved the cryotolerance of resulting blastocysts. While the findings are suggestive of a lipolytic action, further studies are required to clarify the contributions of lipid metabolism and oxidative mechanisms to the observed effects of the L-carnitine treatment. PMID:28302936

  13. Influence of co-culture with denuded oocytes during in vitro maturation on fertilization and developmental competence of cumulus-enclosed porcine oocytes in a defined system.

    Science.gov (United States)

    Appeltant, Ruth; Somfai, Tamás; Kikuchi, Kazuhiro; Maes, Dominiek; Van Soom, Ann

    2016-04-01

    Co-culture of cumulus-oocyte complexes (COCs) with denuded oocytes (DOs) during in vitro maturation (IVM) was reported to improve the developmental competence of oocytes via oocyte-secreted factors in cattle. The aim of the present study was to investigate if addition of DOs during IVM can improve in vitro fertilization (IVF) and in vitro culture (IVC) results for oocytes in a defined in vitro production system in pigs. The maturation medium was porcine oocyte medium supplemented with gonadotropins, dbcAMP and β-mercaptoethanol. Cumulus-oocyte complexes were matured without DOs or with DOs in different ratios (9 COC, 9 COC+16 DO and 9 COC+36 DO). Consequently; oocytes were subjected to IVF as intact COCs or after denudation to examine if DO addition during IVM would affect cumulus or oocyte properties. After fertilization, penetration and normal fertilization rates of zygotes were not different between all tested groups irrespective of denudation before IVF. When zygotes were cultured for 6 days, no difference could be observed between all treatment groups in cleavage rate, blastocyst rate and cell number per blastocyst. In conclusion, irrespective of the ratio, co-culture with DOs during IVM did not improve fertilization parameters and embryo development of cumulus-enclosed porcine oocytes in a defined system. © 2015 Japanese Society of Animal Science.

  14. Morphologic comparison of ovulated and in vitro-matured porcine oocytes, with particular reference to polyspermy after in vitro fertilization.

    Science.gov (United States)

    Wang, W H; Abeydeera, L R; Prather, R S; Day, B N

    1998-03-01

    This study was conducted to evaluate morphologic differences in pig oocytes matured in vivo and in vitro, with particular reference to the potential relationship between oocyte morphology and the occurrence of polyspermy after in vitro fertilization (IVF). In vivo-matured oocytes were surgically recovered from the oviducts of gilts with ovulated follicles on day 2 of estrus, and in vitro-matured oocytes were obtained by culturing follicular oocytes in a oocyte maturation system that has resulted previously in production of live offspring following IVF. Comparisons were made of the cytoplasm density, the diameter of oocytes with or without zona pellucida (ZP), the thickness of the ZP, the size of the perivitelline space (PVS), ZP dissolution time, and cortical granule (CG) distribution before IVF, and CG exocytosis and polyspermic penetration after IVF. Oviductal oocytes have clear areas in the cytoplasm cortex, while in vitro-matured oocytes have very dense cortex. The diameter of ovulated oocytes with ZPs was significantly (P Polyspermy rate was significantly (P polyspermy in pig oocytes.

  15. Factors affecting efficiency of introducing foreign DNA and RNA into parthenogenetic or in vitro-fertilized porcine eggs by cytoplasmic microinjection.

    Science.gov (United States)

    Liu, Shuai; Liu, XiaoQun; Huang, HaiYan; Liu, QingYou; Su, XiaoPing; Zhu, Peng; Li, HongLi; Cui, KuiQing; Xie, BingKun; Shi, DeShun

    2016-08-01

    Cytoplasmic microinjection (CI) of foreign gene into in vivo fertilized zygotes has emerged as a useful tool for transgenic pig production. In the current study, we investigated factors affecting transgenic efficiency and developmental potential of parthenogenetic (PA) and in vitro-fertilized (IVF) porcine embryos produced by CI. These factors included adding of RNase inhibitor, DNA or RNA concentration, injection time, and different structures of plasmids. Our results showed that adding of 1-4 U/μL of RNase inhibitor did not have negative effect on development potential of CI-PA embryos, and RNase inhibitor injection significantly increased EGFP expressing rate of CI-PA embryos. High injection DNA concentration and long injection interval after PA significantly reduced blastocyst formation. Different molecular structures such as DNA or RNA affected CI-PA embryos development, and RNA had little harmful effect on pig's early embryonic development. EGFP expression rate of CI-IVF embryos was improved following the increase of foreign DNA concentration, but blastocyst formation rate was decreased. Injection time after IVF did not show any significant difference on embryonic development, but longer interval resulted in a significantly lower EGFP expressing rate. Cas9 mRNA and myostatin (GDF-8) sgRNA co-injection indicated that the mutation rate of CI-IVF group was significantly higher than that of CI-PA. The CI-IVF-generated embryos were then transferred to six recipient pigs, but no live piglets were obtained. The following pronuclear formation assessment showed more than 76.1% IVF zygotes were polyspermy. These results demonstrate that CI-PA and CI-IVF were effective methods for production of transgenic pig embryos. However, polyspermic fertilization and poor quality of porcine IVF blastocysts are still the main problem of resulting in pregnancy failure.

  16. A new rolling culture-based in vitro fertilization system capable of reducing polyspermy in porcine oocytes.

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    Kitaji, Hideki; Ookutsu, Shoji; Sato, Masahiro; Miyoshi, Kazuchika

    2015-05-01

    The high incidence of polyspermy is one of the major obstacles during in vitro fertilization (IVF) in pigs. To overcome this, we developed a novel IVF method, which involves constant rotation. Oocytes matured in vitro were mixed with spermatozoa (0.2 × 10(5) sperm/mL) in an IVF medium (200 μL) using a 200 μL PCR tube. This tube was then rotated at 1 rpm for 6 h at 38.5°C in a rotation mixer (experimental group). A second PCR tube was simultaneously cultured without rotation (control group). The rate of polyspermy was evaluated 12 h after insemination and was significantly (P < 0.05; 21.0% vs. 48.3%) lower in the experimental group than in the control group. Sperm penetration rate was similar in oocytes from the experimental and control groups (75.2% vs. 83.1%). However, monospermic fertilization rate of the oocytes was significantly (P < 0.05; 44.8% vs. 21.2%) higher in the experimental group than in the control group. Furthermore, the rate of blastocyst formation (30.1% vs. 20.8%) increased in the experimental group, as compared to the control group. This present system will contribute to increase the efficacy of blastocyst production through reduction of polyspermic penetration. © 2014 Japanese Society of Animal Science.

  17. 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

    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...

  18. 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

    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...

  19. Prediction of porcine male fertility

    NARCIS (Netherlands)

    Broekhuijse, M.L.W.J.

    2012-01-01

    Life starts with fertilisation. Variation in fertility is caused by both farm and sow related parameters and boar and semen related parameters. Therefore, achieving high fertility is not obvious. Predominantly, artificial insemination (AI) is used for breeding pigs. The advantage of AI is that you

  20. The Efficient Derivation of Trophoblast Cells from Porcine In Vitro Fertilized and Parthenogenetic Blastocysts and Culture with ROCK Inhibitor Y-27632.

    Directory of Open Access Journals (Sweden)

    Dongxia Hou

    Full Text Available Trophoblasts (TR are specialized cells of the placenta and play an important role in embryo implantation. The in vitro culture of trophoblasts provided an important tool to investigate the mechanisms of implantation. In the present study, porcine trophoblast cells were derived from pig in vitro fertilized (IVF and parthenogenetically activated (PA blastocysts via culturing in medium supplemented with KnockOut serum replacement (KOSR and basic fibroblast growth factor (bFGF on STO feeder layers, and the effect of ROCK (Rho-associated coiled-coil protein kinases inhibiter Y-27632 on the cell lines culture was tested. 5 PA blastocyst derived cell lines and 2 IVF blastocyst derived cell lines have been cultured more than 20 passages; one PA cell lines reached 110 passages without obvious morphological alteration. The derived trophoblast cells exhibited epithelium-like morphology, rich in lipid droplets, and had obvious defined boundaries with the feeder cells. The cells were histochemically stained positive for alkaline phosphatase. The expression of TR lineage markers, such as CDX2, KRT7, KRT18, TEAD4, ELF5 and HAND1, imprinted genes such as IGF2, PEG1 and PEG10, and telomerase activity related genes TERC and TERF2 were detected by immunofluorescence staining, reverse transcription PCR and quantitative real-time PCR analyses. Both PA and IVF blastocysts derived trophoblast cells possessed the ability to differentiate into mature trophoblast cells in vitro. The addition of Y-27632 improved the growth of both PA and IVF blastocyst derived cell lines and increased the expression of trophoblast genes. This study has provided an alternative highly efficient method to establish trophoblast for research focused on peri-implantation and placenta development in IVF and PA embryos.

  1. The effect of FF-MAS on porcine cumulus-oocyte complex maturation, fertilization and pronucleus formation in vitro

    DEFF Research Database (Denmark)

    Færge, Inger; Strejcek, Frantisek; Laurincik, Jozef

    2006-01-01

    Follicular fluid meiosis-activating sterol (FF-MAS) has been isolated from the follicular fluid (FF) of several species including man. FF-MAS increases the quality of in vitro oocyte maturation, and thus the developmental potential of oocytes exposed to FF-MAS during in vitro maturaion is improved...

  2. The quality after culture in vitro or in vivo of porcine oocytes matured and fertilized in vitro and their ability to develop to term.

    Science.gov (United States)

    Nakamura, Yoshiyuki; Tajima, Sigeyuki; Kikuchi, Kazuhiro

    2017-12-01

    The quality of porcine blastocysts produced in vitro is poor in comparison with those that develop in vivo. We examined the quality of in vitro-matured and fertilized (IVM/IVF) oocytes, their abilities to develop to blastocysts under in vivo and in vitro conditions, and the potential of the embryos to develop to term after transfer. IVM/IVF oocytes were either transferred and the embryos recovered on Days 5 and 6 (100% and 87.5%, respectively) ('ET-vivo' embryos), or cultured in vitro for 5 or 6 days ('IVC' embryos). The proportion of blastocysts differed significantly between the two groups on Day 5 (20.6% and 8.0%, respectively), but not on Day 6 (23.8% and 21.2%, respectively). The mean number of cells in ET-vivo blastocysts on Days 5 or 6 was significantly higher (72.8 and 78.7, respectively) than that in IVC blastocysts (22.1 and 39.7, respectively). When IVM/IVF oocytes and IVC blastocysts on Day 6 were transferred, all (three and three, respectively) developed to piglets (16 and 16, respectively), without any difference in the rates of development to term (2.1% and 2.6%, respectively). These data suggest that, although blastocyst production differs between the two culture conditions, IVM/IVF oocytes possess the same ability to develop to term. © 2017 Japanese Society of Animal Science.

  3. A new tool for in vitro culture of porcine eggs | Hua | African Journal ...

    African Journals Online (AJOL)

    A new tool for in vitro culture of porcine eggs. ... are needed for the production of porcine embryos for in vitro fertilization or somatic cell nuclear transfer (SCNT). ... rates with two media were not significantly different (77.7 and 72.4%, P﹤ 0.05 ).

  4. IVF-In Vitro Fertilization.

    Science.gov (United States)

    Kieffer, George H.

    1980-01-01

    Issues surrounding the controversial topic of in vitro fertilization and artificial manipulation of reproduction are discussed. The author examines the moral and ethical implications and presents results of a survey of various religious groups. (SA)

  5. Antihyaluronidase action of ellagic acid effectively prevents polyspermy as a result of suppression of the acrosome reaction induced by sperm-zona interaction during in vitro fertilization of porcine oocytes.

    Science.gov (United States)

    Tokeshi, Isao; Yoshimoto, Teppei; Muto, Norio; Nakamura, Satoshi; Ashizawa, Koji; Nakada, Tadashi; Tatemoto, Hideki

    2007-08-01

    The present study was conducted to examine the effects of three tannin relatives (tannic acid, TA; gallic acid, GA; and ellagic acid, EA) on antihyaluronidase and reactive oxygen species (ROS) scavenging activity, in vitro fertilization (IVF) parameters, and the acrosome reaction (AR) induced by sperm-zona interaction. Among the three tannin relatives, TA and EA showed the strongest potency for blocking the hyaluronidase activity of boar sperm, with concentration-dependent inhibition over the range of 2-10 microg/ml. In contrast, ROSs were effectively scavenged by TA and GA, but not EA. When cumulus-free oocytes were inseminated in IVF medium containing 5 microg/ml of the tannin relatives, polyspermy was significantly reduced by TA and EA (32 and 29%, respectively) compared with oocytes treated with or without GA (51 and 69%, respectively) under conditions that maintained a high sperm penetration rate (Ppolyspermy by suppression of AR functionality induced by sperm-zona interaction and that hyaluronidase intervention is therefore required during porcine IVF.

  6. Effect of Dipeptides on Maturation, Fertilization and Subsequent Embryonic Development of Porcine Oocytes

    Directory of Open Access Journals (Sweden)

    K. M. A. Tareq

    2013-04-01

    Full Text Available The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM-199 supplemented with 10% (v/v porcine follicular fluid, modified Tyrode’s albumin lactate pyruvate (mTALP medium, and modified North Carolina State University (mNCSU-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln+L-glycyl-L-glutamine (GlyGln significantly (p<0.05 improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U-glucose when compared to the control group and other treatment groups. Additionally, 2–4 cell, 8–16 cell, morula and blastocyst development increased significantly (p<0.05 following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media.

  7. Pro-apoptotic Effect of Pifithrin-α on Preimplantation Porcine Fertilized Embryo Development

    Directory of Open Access Journals (Sweden)

    Brendan Mulligan

    2012-12-01

    Full Text Available The aim of this study was to investigate the impact of a reported p53 inhibitor, pifithrin-α (PFT-α, on preimplantation porcine in vitro fertilized (IVF embryo development in culture. Treatment of PFT-α was administered at both early (0 to 48 hpi, and later stages (48 to 168 hpi of preimplantation development, and its impact upon the expression of five genes related to apoptosis (p53, bak, bcl-xL, p66Shc and caspase3, was assessed in resulting d 7 blastocysts, using real-time quantitative PCR. Total cell numbers, along with the number of apoptotic nuclei, as detected by the in situ cell death detection assay, were also calculated on d 7 in treated and non-treated control embryos. The results indicate that PFT-α, when administered at both early and later stages of porcine IVF embryo development, increases the incidence of apoptosis in resulting blastocysts. When administered at early cleavage stages, PFT-α treatment was shown to reduce the developmental competence of porcine IVF embryos, as well as reducing the quality of resulting blastocysts in terms of overall cell numbers. In contrast, at later stages, PFT-α administration resulted in marginally increased blastocyst development rates amongst treated embryos, but did not affect cell numbers. However, PFT-α treatment induced apoptosis and apoptotic related gene expression, in all treated embryos, irrespective of the timing of treatment. Our results indicate that PFT-α may severely compromise the developmental potential of porcine IVF embryos, and is a potent apoptotic agent when placed into porcine embryo culture media. Thus, caution should be exercised when using PFT-α as a specific inhibitor of p53 mediated apoptosis, in the context of porcine IVF embryo culture systems.

  8. Effects of griseofulvin on in vitro porcine oocyte maturation and embryo development.

    Science.gov (United States)

    Miao, Yi-Liang; Zhang, Xia; Zhao, Jian-Guo; Spate, Lee; Zhao, Ming-Tao; Murphy, Clifton N; Prather, Randall S; Sun, Qing-Yuan; Schatten, Heide

    2012-08-01

    Griseofulvin is an orally administered antifungal drug that affects microtubule formation in vitro and interferes with microtubule dynamics in vivo as clearly shown for mitotic cells in several cell systems. This article reports the effects of griseofulvin on in vitro maturation of porcine oocytes and subsequent effects on embryo development. Our results revealed a concentration-dependent effect on meiotic spindles with 20-40 μM griseofulvin affecting oocyte maturation, and 40 μM affecting fertilization and embryo development. These concentrations of griseofulvin did not affect mitochondrial and cortical granule distribution that also depend on microtubule and cytoskeletal functions during oocyte maturation. Specific effects on the meiotic spindle included spindle disorganization and aberrant chromosome separation displayed as prominent chromosome clusters in oocytes treated with 40 μM griseofulvin. These results strongly suggested that griseofulvin affected porcine oocyte in vitro maturation and following embryo development by disturbing microtubule dynamics. Copyright © 2012 Wiley Periodicals, Inc.

  9. Combination effects of epidermal growth factor and glial cell line-derived neurotrophic factor on the in vitro developmental potential of porcine oocytes

    DEFF Research Database (Denmark)

    Valleh, Mehdi Vafaye; Rasmussen, Mikkel Aabech; Hyttel, Poul

    2016-01-01

    The developmental potential of in vitro matured porcine oocytes is still lower than that of oocytes matured and fertilized in vivo. Major problems that account for the lower efficiency of in vitro production include the improper nuclear and cytoplasmic maturation of oocytes. With the aim of impro...

  10. Human rights to in vitro fertilization.

    Science.gov (United States)

    Zegers-Hochschild, Fernando; Dickens, Bernard M; Dughman-Manzur, Sandra

    2013-10-01

    The Inter-American Court of Human Rights (the Court) has ruled that the Supreme Court of Costa Rica's judgment in 2000 prohibiting in vitro fertilization (IVF) violated the human right to private and family life, the human right to found and raise a family, and the human right to non-discrimination on grounds of disability, financial means, or gender. The Court's conclusions of violations contrary to the American Convention on Human Rights followed from its ruling that, under the Convention, in vitro embryos are not "persons" and do not possess a right to life. Accordingly, the prohibition of IVF to protect embryos constituted a disproportionate and unjustifiable denial of infertile individuals' human rights. The Court distinguished fertilization from conception, since conception-unlike fertilization-depends on an embryo's implantation in a woman's body. Under human rights law, legal protection of an embryo "from conception" is inapplicable between its creation by fertilization and completion of its implantation in utero. © 2013.

  11. In vitro fertilization with preimplantation genetic screening

    NARCIS (Netherlands)

    Mastenbroek, Sebastiaan; Twisk, Moniek; van Echten-Arends, Jannie; Sikkema-Raddatz, Birgit; Korevaar, Johanna C.; Verhoeve, Harold R.; Vogel, Niels E. A.; Arts, Eus G. J. M.; de Vries, Jan W. A.; Bossuyt, Patrick M.; Buys, Charles H. C. M.; Heineman, Maas Jan; Repping, Sjoerd; van der Veen, Fulco

    2007-01-01

    BACKGROUND: Pregnancy rates in women of advanced maternal age undergoing in vitro fertilization (IVF) are disappointingly low. It has been suggested that the use of preimplantation genetic screening of cleavage-stage embryos for aneuploidies may improve the effectiveness of IVF in these women.

  12. [Coronary angiography of in vitro porcine heart using MSCT].

    Science.gov (United States)

    Wan, Lei; Telet, Siy It; Wei, Hua; Ying, Chong-Liang; Wang, Ya-hui; Deng, Kai-fei; Zou, Dong-Hua; Li, Zheng-done; Zhu, Guang-You

    2014-10-01

    To establish standardized methods and parameters of the isolated heart coronary angiography through the experiment of in vitro porcine heart by MSCT. Based on different perfusion volume (50, 60 and 70 mL) and different perfusion-imaging time (5, 10 and 20 min), the in vitro porcine coronary artery was injected liposoluble and water-soluble contrast agents using remodel angiography equipment and scanned by MSCT. And the 3D image results were compared. The images were recorded and evaluated by 2 radiologists and analyzed by statistical software. Liposoluble contrast agent affected the images by damaging and infiltrating the fats around the coronary artery, while the water-soluble contrast agent didn't affect the images. The groups with 60 mL or 70 mL perfusion and 5 min perfusion-imaging time had the best images. The suitable parameters of the angiography lay the foundation of postmortem coronary angiography.

  13. Interphase chromosome positioning in in vitro porcine cells and ex vivo porcine tissues

    Directory of Open Access Journals (Sweden)

    Foster Helen A

    2012-11-01

    Full Text Available Abstract Background In interphase nuclei of a wide range of species chromosomes are organised into their own specific locations termed territories. These chromosome territories are non-randomly positioned in nuclei which is believed to be related to a spatial aspect of regulatory control over gene expression. In this study we have adopted the pig as a model in which to study interphase chromosome positioning and follows on from other studies from our group of using pig cells and tissues to study interphase genome re-positioning during differentiation. The pig is an important model organism both economically and as a closely related species to study human disease models. This is why great efforts have been made to accomplish the full genome sequence in the last decade. Results This study has positioned most of the porcine chromosomes in in vitro cultured adult and embryonic fibroblasts, early passage stromal derived mesenchymal stem cells and lymphocytes. The study is further expanded to position four chromosomes in ex vivo tissue derived from pig kidney, lung and brain. Conclusions It was concluded that porcine chromosomes are also non-randomly positioned within interphase nuclei with few major differences in chromosome position in interphase nuclei between different cell and tissue types. There were also no differences between preferred nuclear location of chromosomes in in vitro cultured cells as compared to cells in tissue sections. Using a number of analyses to ascertain by what criteria porcine chromosomes were positioned in interphase nuclei; we found a correlation with DNA content.

  14. Comparison of physiological and in vitro porcine gastric fluid digestion.

    Science.gov (United States)

    Kopper, Randall A; West, Charles M; Helm, Ricki M

    2006-01-01

    In previous studies, the major peanut allergen Ara h 1 was digested in vitro using pepsin and porcine gastric fluid. The results suggested that in vivo gastric digestion of allergen protein can be modeled accurately by peptic hydrolysis in vitro. In the current investigation, studies were designed to follow the gastrointestinal (GI) digestion of peanut allergens under true physiological conditions. In vitro digestion with porcine gastric fluid was compared with actual physiological digestion of peanut allergens in the porcine digestive tract in vivo. Analysis of physiologic digestion was performed in piglets administered a 20-gram bolus of peanut meal followed by periodic sampling and analysis of GI contents. The pH was monitored, and digesta were analyzed by SDS-PAGE and immunoblot analysis. Peanut meal initially neutralized stomach contents to a pH of approximately 7, which was subsequently acidified by HCl secretion within 30 min. Acidification to pH 2-4 resulted in active pepsin digestion of soluble protein in the stomach. Soluble intact protein/allergens were rapidly degraded to pepsin-resistant peptides in the stomach followed by hydrolysis of these fragments in the small intestine. Particulate material was evident in both the stomach and small intestine that could contribute to continued release of peanut allergens Ara h 1, 2 and 3. Porcine gastric digestion of peanut proteins resembles true physiological digestion only under optimal physiologic conditions. Soluble proteins are rapidly digested and insoluble material continues to release IgE-reactive proteins throughout the GI tract. GI digestion of food allergens can play a prominent role when assessing allergens within the context of a food matrix or meal and during the sensitization phase of IgE-mediated allergy.

  15. In vitro manipulation techniques of porcine embryos

    DEFF Research Database (Denmark)

    Liu, Ying; Li, Juan; Løvendahl, Peter

    2015-01-01

    During the last 17 years, considerable advancements have been achieved in the production of pigs, transgenic and non-transgenic, by methods of somatic cell nuclear transfer, in vitro fertilisation, intracytoplasmic sperm injection, microinjection and sperm-mediated gene transfer by artificial...

  16. SNARE protein mediated secretory events in porcine fertilization

    NARCIS (Netherlands)

    Tsai, P.S.|info:eu-repo/dai/nl/304834742

    2010-01-01

    Fertilization is an event that involves highly dynamic intra- and intercellular processes which enable the merge of two gametes with the aim to ultimately form a new organism. Preparative changes are required in both gametes prior to fertilization since freshly ejaculated sperm cells and germinal

  17. BREAST CANCER AND IN VITRO FERTILIZATION PROGRAM

    Directory of Open Access Journals (Sweden)

    A. A. Parokonnaya

    2008-01-01

    Full Text Available Eleven cases of breast cancer (BC diagnosed during or after in-vitro fertilization (IVF are considered. The study group was observed to have cases of familial BC in 18.2% of the patients and BRCA1 gene mutations in 60%. At the same time 45.5% of cases had local- ly advanced and generalized BC, which suggests that there is a tumor at the initiation of a regular IVF cycle. The authors arrive at the conclusion that it is necessary to include a mammologist's examination, breast ultrasonography, and an oncogeneticist's counseling into the package of compulsory measures during IVF.

  18. "Cell Migration" Is the Ontology Group Differentially Expressed in Porcine Oocytes Before and After In Vitro Maturation: A Microarray Approach.

    Science.gov (United States)

    Kranc, Wiesława; Budna, Joanna; Chachuła, Adrian; Borys, Sylwia; Bryja, Artur; Rybska, Marta; Ciesiółka, Sylwia; Sumelka, Ewa; Jeseta, Michał; Brüssow, Klaus P; Bukowska, Dorota; Antosik, Paweł; Bruska, Małgorzata; Nowicki, Michał; Zabel, Maciej; Kempisty, Bartosz

    2017-04-01

    Maturation of cumulus-oocyte complexes (COCs) is crucial for further successful monospermic fertilization, embryo growth, and implantation. All these events are accompanied by proliferation and differentiation of cumulus cells. The migration of COCs to the oviduct after ovulation and the interaction between female gametes and/or embryos with maternal tissues are still poorly recognized on the molecular level. This study was aimed to first demonstrate the mRNA expression profile of cell migration markers during different stages of porcine oocytes maturation and developmental capability in vitro. The COCs were collected from a total of 45 pubertal crossbred Landrace gilts, brilliant cresyl blue (BCB) stained, and analyzed before (n = 150) or after (n = 150) in vitro maturation (IVM). Using the Affymetrix® Porcine Gene 1.1 ST Array, the expression profile of 12,258 porcine transcripts was examined. We found nine genes involved in cell migration mechanisms, that is, PLD1, KIT, LAMA2, MAP3K1, VEGFA, TGFBR3, INSR, TPM1, and RTN4. These genes were upregulated in porcine oocytes before IVM as compared with post-IVM expression analysis. Moreover, important mechanisms of biological interaction between VEGFA-KIT and VEGFA-INSR were also observed. The upregulation and/or downregulation of selected mRNAs expression after microarray assays was checked and approved by real-time quantitative polymerase chain reaction. We suggest that several genes, including LAMA2 or TPM1, encode proteins participating in the formation of the oocyte's protein architecture such as microtubules and kinetochore reorganization. As the expression of all "migration regulatory genes" investigated in this study was significantly upregulated in oocytes before IVM, we conclude that they may contribute to the maturational capability of porcine oocytes. However, migration potency of COCs is not accompanied by achievement of the MII stage by porcine oocytes in vitro. The investigated genes such as PLD

  19. Increased venous thrombosis incidence in pregnancies after in vitro fertilization

    DEFF Research Database (Denmark)

    Hansen, Anette Tarp; Kesmodel, U S; Juul, S

    2014-01-01

    STUDY QUESTION Is venous thrombosis risk increased in pregnancies after in vitro fertilization? SUMMARY ANSWER The venous thrombosis incidence was significantly increased in pregnancies after in vitro fertilization; especially in the first trimester and in the first 6 weeks post-partum. WHAT...... IS KNOWN ALREADY In vitro fertilization without pregnancy is not associated with increased venous thrombosis incidence. STUDY DESIGN, SIZE, DURATION This national register-based cohort study covered the period from 1995 to 2005. PARTICIPANTS/MATERIALS, SETTING, METHODS All Danish pregnancies conceived...... by in vitro fertilization (n = 18 787) were included. Venous thrombosis incidence rates in pregnancies after in vitro fertilization were compared with venous thrombosis incidence rates in reference pregnancies, by calculating incidence rate ratios. MAIN RESULTS AND THE ROLE OF CHANCE In total, 48 cases were...

  20. Ubiquitin C-terminal hydrolase-activity is involved in sperm acrosomal function and anti-polyspermy defense during porcine fertilization.

    Science.gov (United States)

    Yi, Young-Joo; Manandhar, Gaurishankar; Sutovsky, Miriam; Li, Rongfeng; Jonáková, Vera; Oko, Richard; Park, Chang-Sik; Prather, Randall S; Sutovsky, Peter

    2007-11-01

    The 26S proteasome, which is a multi-subunit protease with specificity for substrate proteins that are postranslationally modified by ubiquitination, has been implicated in acrosomal function and sperm-zona pellucida (ZP) penetration during mammalian fertilization. Ubiquitin C-terminal hydrolases (UCHs) are responsible for the removal of polyubiquitin chains during substrate priming for proteasomal proteolysis. The inhibition of deubiquitination increases the rate of proteasomal proteolysis. Consequently, we have hypothesized that inhibition of sperm acrosome-borne UCHs increases the rate of sperm-ZP penetration and polyspermy during porcine in vitro fertilization (IVF). Ubiquitin aldehyde (UA), which is a specific nonpermeating UCH inhibitor, significantly (P polyspermy during porcine IVF and reduced (P polyspermy during IVF, consistent with the UA-induced polyspermy surge. In the oocyte, UCHL3 was primarily associated with the meiotic spindle. Sperm-borne UCHL3 was localized to the acrosomal surface and coimmunoprecipitated with a peripheral acrosomal membrane protein, spermadhesin AQN1. Recombinant UCHs, UCHL3, and isopeptidase T reduced polyspermy when added to the fertilization medium. UCHL1 was detected in the oocyte cortex but not on the sperm surface, and was partially degraded 6-8 h after fertilization. Enucleated oocyte-somatic cell electrofusion caused polarized redistribution of cortical UCHL1. We conclude that sperm-acrosomal UCHs are involved in sperm-ZP interactions and antipolyspermy defense. Modulation of UCH activity could facilitate the management of polyspermy during IVF and provide insights into male infertility.

  1. The effect of oviductal epithelial cell co-culture during in vitro maturation on sow oocyte morphology, fertilization and embryo development

    NARCIS (Netherlands)

    Kidson, A.; Schoevers, E.; Langendijk, P.; Verheijden, J.; Colenbrander, B.; Bevers, M.

    2003-01-01

    In vitro embryo production in the sow is challenged by poor cytoplasmic maturation, low sperm penetration and low normal fertilization, leading to the development of poor quality blastocysts containing a small number of nuclei. In prepubertal gilt oocytes, the presence of porcine oviductal

  2. The secretions of oviduct epithelial cells increase the equine in vitro fertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?

    OpenAIRE

    Mugnier, Sylvie; Kervella, Morgane; Douet, C?cile; Canepa, Sylvie; Pascal, G?raldine; Deleuze, Stefan; Duchamp, Guy; Monget, Philippe; Goudet, Ghyl?ne

    2009-01-01

    Abstract Background Oviduct epithelial cells (OEC) co-culture promotes in vitro fertilization (IVF) in human, bovine and porcine species, but no data are available from equine species. Yet, despite numerous attempts, equine IVF rates remain low. Our first aim was to verify a beneficial effect of the OEC on equine IVF. In mammals, oviductal proteins have been shown to interact with gametes and play a role in fertilization. Thus, our second aim was to identify the proteins involved in fertiliza...

  3. "Bone Development" Is an Ontology Group Upregulated in Porcine Oocytes Before In Vitro Maturation: A Microarray Approach.

    Science.gov (United States)

    Budna, Joanna; Bryja, Artur; Celichowski, Piotr; Kranc, Wiesława; Ciesiółka, Sylwia; Borys, Sylwia; Rybska, Marta; Kolecka-Bednarczyk, Agata; Jeseta, Michal; Bukowska, Dorota; Antosik, Paweł; Brüssow, Klaus P; Bruska, Małgorzata; Nowicki, Michał; Zabel, Maciej; Kempisty, Bartosz

    2017-08-01

    Mammalian cumulus-oocyte complexes (COCs) reach full developmental capability during folliculogenesis and oogenesis. It is well recognized that only gametes achieving MII stage after in vivo or in vitro maturation (IVM) are successfully fertilized by a single spermatozoon. Although the process of oocyte nuclear and/or cytoplasmic maturation in pigs is well determined, there exist many differences that promote these processes in vivo and in vitro. Therefore, this study aimed to investigate the differences in RNA expression profiles between porcine oocytes before and after IVM using microarray and real-time quantitative polymerase chain reaction (RT-qPCR) assays. Experiments were performed on oocytes isolated from 55 pubertal crossbred Landrace gilts. The oocytes were analyzed both before and after IVM and only Brilliant Cresyl Blue (BCB)-positive gametes were used for subsequent microarray analysis (Affymetrix) and RT-qPCR analysis. The microarray assay, which measures expression of 12,258 transcripts, revealed 419 differentially expressed transcripts in porcine oocytes, from which 379 were downregulated and 40 were upregulated before IVM compared to those analyzed after IVM. After DAVID analysis, we found eight different transcripts, including IHH, BMP1, WWTR1, CHRDL1, KLF10, EIF2AK3, MMP14, and STC1. Their expression is related to the "bone development" ontology group and was further subjected to hierarchical clusterization. Using RT-qPCR analysis, we confirmed the results of the microarray assay, showing increased expression of the eight genes in oocytes before IVM compared to oocytes after maturation in vitro. It has been suggested that "bone development" belongs to one ontological group involving genes substantially upregulated in porcine oocytes before IVM. We suggest that the gamete mRNA expression profile before IVM may comprise stored transcripts, which are templates for protein biosynthesis following fertilization. We also hypothesize that these mRNAs may

  4. X-linked gene transcription patterns in female and male in vivo, in vitro and cloned porcine individual blastocysts.

    Directory of Open Access Journals (Sweden)

    Chi-Hun Park

    Full Text Available To determine the presence of sexual dimorphic transcription and how in vitro culture environments influence X-linked gene transcription patterns in preimplantation embryos, we analyzed mRNA expression levels in in vivo-derived, in vitro-fertilized (IVF, and cloned porcine blastocysts. Our results clearly show that sex-biased expression occurred between female and male in vivo blastocysts in X-linked genes. The expression levels of XIST, G6PD, HPRT1, PGK1, and BEX1 were significantly higher in female than in male blastocysts, but ZXDA displayed higher levels in male than in female blastocysts. Although we found aberrant expression patterns for several genes in IVF and cloned blastocysts, similar sex-biased expression patterns (on average were observed between the sexes. The transcript levels of BEX1 and XIST were upregulated and PGK1 was downregulated in both IVF and cloned blastocysts compared with in vivo counterparts. Moreover, a remarkable degree of expression heterogeneity was observed among individual cloned embryos (the level of heterogeneity was similar in both sexes but only a small proportion of female IVF embryos exhibited variability, indicating that this phenomenon may be primarily caused by faulty reprogramming by the somatic cell nuclear transfer (SCNT process rather than in vitro conditions. Aberrant expression patterns in cloned embryos of both sexes were not ameliorated by treatment with Scriptaid as a potent HDACi, although the blastocyst rate increased remarkably after this treatment. Taken together, these results indicate that female and male porcine blastocysts produced in vivo and in vitro transcriptional sexual dimorphisms in the selected X-linked genes and compensation of X-linked gene dosage may not occur at the blastocyst stage. Moreover, altered X-linked gene expression frequently occurred in porcine IVF and cloned embryos, indicating that X-linked gene regulation is susceptible to in vitro culture and the SCNT process

  5. Porcine oocyte maturation in vitro: role of cAMP and oocyte-secreted factors – A practical approach

    Science.gov (United States)

    APPELTANT, Ruth; SOMFAI, Tamás; MAES, Dominiek; VAN SOOM, Ann; KIKUCHI, Kazuhiro

    2016-01-01

    Polyspermy or the penetration of more than one sperm cell remains a problem during porcine in vitro fertilization (IVF). After in vitro culture of porcine zygotes, only a low percentage of blastocysts develop and their quality is inferior to that of in vivo derived blastocysts. It is unknown whether the cytoplasmic maturation of the oocyte is sufficiently sustained in current in vitro maturation (IVM) procedures. The complex interplay between oocyte and cumulus cells during IVM is a key factor in this process. By focusing on this bidirectional communication, it is possible to control the coordination of cumulus expansion, and nuclear and cytoplasmic maturation during IVM to some extent. Therefore, this review focuses on the regulatory mechanisms between oocytes and cumulus cells to further the development of new in vitro embryo production (IVP) procedures, resulting in less polyspermy and improved oocyte developmental potential. Specifically, we focused on the involvement of cAMP in maturation regulation and function of oocyte-secreted factors (OSFs) in the bidirectional regulatory loop between oocyte and cumulus cells. Our studies suggest that maintaining high cAMP levels in the oocyte during the first half of IVM sustained improved oocyte maturation, resulting in an enhanced response after IVF and cumulus matrix disassembly. Recent research indicated that the addition of OSFs during IVM enhanced the developmental competence of small follicle-derived oocytes, which was stimulated by epidermal growth factor (EGF) via developing EGF-receptor signaling. PMID:27349308

  6. Porcine oocyte maturation in vitro: role of cAMP and oocyte-secreted factors - A practical approach.

    Science.gov (United States)

    Appeltant, Ruth; Somfai, Tamás; Maes, Dominiek; VAN Soom, Ann; Kikuchi, Kazuhiro

    2016-10-18

    Polyspermy or the penetration of more than one sperm cell remains a problem during porcine in vitro fertilization (IVF). After in vitro culture of porcine zygotes, only a low percentage of blastocysts develop and their quality is inferior to that of in vivo derived blastocysts. It is unknown whether the cytoplasmic maturation of the oocyte is sufficiently sustained in current in vitro maturation (IVM) procedures. The complex interplay between oocyte and cumulus cells during IVM is a key factor in this process. By focusing on this bidirectional communication, it is possible to control the coordination of cumulus expansion, and nuclear and cytoplasmic maturation during IVM to some extent. Therefore, this review focuses on the regulatory mechanisms between oocytes and cumulus cells to further the development of new in vitro embryo production (IVP) procedures, resulting in less polyspermy and improved oocyte developmental potential. Specifically, we focused on the involvement of cAMP in maturation regulation and function of oocyte-secreted factors (OSFs) in the bidirectional regulatory loop between oocyte and cumulus cells. Our studies suggest that maintaining high cAMP levels in the oocyte during the first half of IVM sustained improved oocyte maturation, resulting in an enhanced response after IVF and cumulus matrix disassembly. Recent research indicated that the addition of OSFs during IVM enhanced the developmental competence of small follicle-derived oocytes, which was stimulated by epidermal growth factor (EGF) via developing EGF-receptor signaling.

  7. In-vitro Fertilization, Gamete Donation and Surrogacy: Perceptions ...

    African Journals Online (AJOL)

    AJRH Managing Editor

    ART) offered in Nigeria include in-vitro fertilization (IVF), gamete donation and surrogacy. This cross-sectional questionnaire study aimed at assessing the acceptability of ART to women seeking infertility treatment at the University College Hospital ...

  8. In vitro fertilization of banana prawn Penaeus merguiensis de Man

    Digital Repository Service at National Institute of Oceanography (India)

    Nair, S.R.S.

    Successful in Vitro fertilization of the commercially important prawn, Penaeus merguiensis has been achieved. Suspensions of sperms and ova were mixed and agitated for five minutes. Healthy nauplii were observed on following day and developed...

  9. [Ectopic pregnancy following in vitro fertilization and embryo transfer].

    Science.gov (United States)

    Sudik, R; Fliess, F R; Bernt, W D; Meissner, J; Kunkel, S

    1984-01-01

    A report is given about one case of ectopic pregnancy after in vitro fertilization and transfer of three embryos. Possible causes of ectopic pregnancies following embryo transfer and conclusions are discussed.

  10. In-Vitro Fertilization Practice: Awareness and Perceptions ...

    African Journals Online (AJOL)

    BACKGROUND: In-vitro fertilization(IVF) is a procedure that is one of the options in managing infertility. It involves fertilizing the egg from the female with the spermatozoa from the male outside the human body. It is a common practice in the developed world. However it is still a new phenomenon in the developing countries.

  11. Alternative approaches to ovarian stimulation for in vitro fertilization

    NARCIS (Netherlands)

    D. de Jong (Diederick)

    2003-01-01

    markdownabstract__Abstract__ In vitro fertilization (IVF), literally meaning ‘fertilization in glass’, is characterized by co-culture of aspired oocytes with spermatozoa and subsequent transfer of the embryo(s) into the uterine cavity. The original indication for this treatment was

  12. Beneficial effects of serum supplementation during in vitro production of porcine embryos on their ability to survive cryopreservation by open pulled straw vitrification.

    Science.gov (United States)

    Men, Hongsheng; Agca, Yuksel; Critser, Elizabeth S; Critser, John K

    2005-10-01

    The ability of porcine blastocysts produced in vitro, in the presence or absence of serum, to survive cryopreservation was investigated in this experiment. Porcine oocytes were matured, fertilized and cultured in vitro using serum-free culture systems. Starting at Day 4 of in vitro embryo culture (Day 0 = fertilization), the culture medium was supplemented with 10% fetal bovine serum (FBS). Embryos were cultured under these conditions until Day 6. Embryos cultured with only BSA supplementation served as serum-free controls. Day 6 blastocysts and expanded blastocysts of excellent quality were vitrified using the open pulled straw method. After warming, blastocysts were cultured in the presence of 10% FBS for an additional 18 h to recover. Portions of blastocysts from both groups, without cryopreservation, were also cultured under the same conditions to serve as non-vitrified controls. To further investigate the influence of FBS on the quality of embryos produced, the total cell numbers in Day 6 blastocysts from both groups were compared. In addition, the ratio of viable to total cells in fully recovered blastocysts at each group was examined. Blastocysts produced in the presence of FBS had an increased ability to survive cryopreservation and also had a higher cell number compared to those produced in serum-free systems (P vitro production of porcine embryos is beneficial to the ability of a blastocyst to survive cryopreservation.

  13. Identification of maturation and protein synthesis related proteins from porcine oocytes during in vitro maturation

    Directory of Open Access Journals (Sweden)

    Seo Kang

    2011-06-01

    Full Text Available Abstract Background In vitro maturation (IVM of mammalian oocytes is divided into the GV (germinal vesicle stage, MI (metaphase I stage and MII (metaphase II stage stages, and only fully mature oocytes have acquired the ability to be fertilized and initiate zygotic development. These observations have been mostly based on morphological evaluations, but the molecular events governing these processes are not fully understood. The aim of the present study was to better understand the processes involved in the molecular regulation of IVM using 2-DE analysis followed by mass spectrometry to identify proteins that are differentially expressed during oocyte IVM. Result A total of 16 up-regulated and 12 down-regulated proteins were identified. To investigate the IVM process, we specifically focused on the proteins that were up-regulated during the MII stage when compared with the GV stage, which included PRDX 2, GST, SPSY, myomegalin, PED4D, PRKAB 1, and DTNA. These up-regulated proteins were functionally involved in redox regulation and the cAMP-dependent pathway, which are essential for the intracellular signaling involved in oocyte maturation. Interestingly, the PDE4D and its partner, myomegalin, during the MII stage was consistently confirmed up-regulation by western blot analyses. Conclusion These results could be used to better understand some aspects of the molecular mechanisms underlying porcine oocyte maturation. This study identified some regulatory proteins that may have important roles in the molecular events involved in porcine oocyte maturation, particularly with respect to the regulation of oocyte meiotic resumption, MII arrest and oocyte activation. In addition, this study may have beneficial applications not only to basic science with respect to the improvement of oocyte culture conditions but also to mammalian reproductive biotechnology with potential implications.

  14. Factors affecting the development of in vitro fertilization in camelids

    Directory of Open Access Journals (Sweden)

    Trasorras VL

    2016-12-01

    Full Text Available In any program of in vitro embryo production, the ultimate goal is to develop high quality embryos being able to get a normal pregnancy and finally resulting in the birth of a healthy offspring, goal not reach yet in camelids. The application of assisted reproductive techniques, such as in vitro fertilization and subsequent in vitro embryo culture, can extend the knowledge of early embryonic development and make possible the increase of the population of genetically superior animals.

  15. Optical clearing of porcine skin tissue in vitro studied by Raman microspectroscopy

    Science.gov (United States)

    Huang, Deqiu; Zhang, Wen; Zhong, Huiqing; Xiong, Honglian; Guo, Xi; Guo, Zhouyi

    2012-01-01

    In present work, we studied the effect of optical clearing on porcine skin in vitro with glycerol by Raman microspectroscopy, denoted as RM, at various time intervals of 0, 15, 30, 45, 60, and 75 min respectively. The results showed that the addition of glycerol significantly improved the depth of RM measurement, and enhanced the recovery of skin tissue Raman spectra that were not overlapped with the glycerol Raman spectra over time. Moreover, it was found that the Raman signals resembled the native spectrum of the molecules in porcine skin with a negligible frequency shift. Furthermore, we evaluated the extent of optical clearing in porcine skin by utilizing various concentrations of 40%, 60%, and 80% glycerol solution. The results demonstrated that with the increase of concentration of glycerol, the optical clearing of porcine skin was much improved.

  16. Perspectives on access to in vitro fertilization in Portugal

    Directory of Open Access Journals (Sweden)

    S. Silva

    2012-04-01

    Full Text Available OBJECTIVE: To analyze users' reasons for choosing in vitro fertilization treatment in public or private services and to identify their suggestions for improving fertility treatment. METHODS: A qualitative study using an interpretative approach was conducted. Fifteen semi-structured interviews were conducted with patients undergoing in vitro fertilization treatment (nine women, one man and five couples at home or at their workplace in the districts of Viana do Castelo, Braga, Porto and Lisbon, Portugal, between July 2005 and February 2006. RESULTS: Users evaluated access to in vitro fertilization treatment in public and private services based mainly on their individual experiences and called for more access to less costly, faster and friendlier care with suitable facilities, appropriate time management and caring medical providers. These perceptions were also associated with views on the need for fighting stigmatization of infertility, protecting children's rights and guaranteeing sustainability of health care system. Interviewees sought to balance reduced waiting time and more attentive care with costs involved. The choice of services depended on the users' purchase power and place of residence and availability of attentive care. CONCLUSIONS: Current national policies on in vitro fertilization treatment meet user's demands of promoting access to, and quality, availability and affordability of in vitro fertilization treatment. However, their focus on legal regulation and technical-scientific aspects contrasts with the users' emphasis on reimbursement, insurance coverage and focus on emotional aspects of the treatment. The study showed these policies should ensure insurance coverage, participation of user representatives in the National Council for Assisted Reproductive Technology, promotion of infertility research and certification of fertility laboratories.

  17. Deleted in malignant brain tumour 1 (DMBT1) is secreted in the oviduct and involved in the mechanism of fertilization in equine and porcine species

    DEFF Research Database (Denmark)

    Ambruosi, Barbara; Accogli, Gianluca; Douet, Cecile

    2013-01-01

    fertilization (IVF) in porcine and equine species that represent divergent IVF models. We first performed IVF after pre-incubation of oocytes with or without oviductal fluid supplemented or not with antibodies directed against DMBT1. We showed that oviductal fluid induces an increase of the monospermic...... fertilization rate, and that this effect is cancelled by the addition of antibodies, in both porcine and equine species. Moreover, pre-incubation of oocytes with recombinant DMBT1 induces an increase of the monospermic fertilization rate in the pig, confirming an involvement of DMBT1 in the fertilization...... in the zona pellucida and cytoplasm of equine and porcine oocytes was observed using immunofluorescence analysis and confocal microscopy. Moreover, we showed an interaction between DMBT1 and porcine spermatozoa using surface plasmon resonance studies. Finally, a bioinformatics and phylogenetic analysis...

  18. In Vitro Fermentation of Porcine Milk Oligosaccharides and Galacto-oligosaccharides Using Piglet Fecal Inoculum

    NARCIS (Netherlands)

    Difilippo, Elisabetta; Pan, Feipeng; Logtenberg, Madelon; Willems, Rianne; Braber, Saskia; Fink-Gremmels, Johanna; Schols, Henk A.; Gruppen, Harry

    2016-01-01

    In this study, the in vitro fermentation by piglet fecal inoculum of galacto-oligosaccharides (GOS) and porcine milk oligosaccharides (PMOs) was investigated to identify possible preferences for individual oligosaccharide structures by piglet microbiota. First, acidic PMOs and GOS with degrees of

  19. In Vitro Fermentation of Porcine Milk Oligosaccharides and Galacto-oligosaccharides Using Piglet Fecal Inoculum

    NARCIS (Netherlands)

    Difilippo, Elisabetta; Pan, Feipeng; Logtenberg, Madelon; Willems, Rianne H A M; Braber, Saskia; Fink-Gremmels, Johanna; Schols, Henk A; Gruppen, Harry

    In this study, the in vitro fermentation by piglet fecal inoculum of galacto-oligosaccharides (GOS) and porcine milk oligosaccharides (PMOs) was investigated to identify possible preferences for individual oligosaccharide structures by piglet microbiota. First, acidic PMOs and GOS with degrees of

  20. In-Vitro Fertilization Practice: Awareness and Perceptions Among ...

    African Journals Online (AJOL)

    vitro fertilization (IVF) is a procedure in which eggs from the woman's ovary are removed, and are mixed with sperm in an ... However it is still a new phenomenon in the developing countries. This is due to the .... rate could be as a result of previous untreated sexually transmitted diseases, post abortion complications and. 10.

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

    African Journals Online (AJOL)

    Assisted reproductive technology (ART) has restored hope to millions of infertile couples globally, with In vitro fertilization (IVF) and embryo transfer offered in virtually all. ART units. This development in medicine lead to the delivery of the first test tube baby named Louise Brown on July 25,. 1978 in Oldham, England after a ...

  2. In-Vitro Fertilization Practice: Awareness and Perceptions Among ...

    African Journals Online (AJOL)

    unprotected sexual intercourse. Africa has the highest rate of infertility, mostly due to tubal occlusion. Paradoxically, however, it records the lowest rate of assisted reproductive technology (ART) treatment. In vitro fertilization (IVF) is a procedure in which eggs from the woman's ovary are removed, and are mixed with sperm in ...

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

    African Journals Online (AJOL)

    The number of embryos transferred after in vitro fertilization (IVF) have been a topic of debate for over a decade now. Due to the risk associated with multiple pregnancy, there has been a global effort at reducing the multiple pregnancy rates to a minimum while maintaining an acceptable level of successful IVF pregnancy ...

  4. Perceptions of Infertility and In Vitro Fertilization Treatment among ...

    African Journals Online (AJOL)

    This study thus explored the perceptions of infertility and in vitro fertilization (IVF) and how to enhance the use of IVF treatment among married couples. This was a cross-sectional survey in Anambra State, Nigeria involving household respondents (married couples) and hospital respondents (couples undergoing infertility ...

  5. Miliary tuberculosis after in vitro fertilization and embryo ...

    African Journals Online (AJOL)

    Background: With the development of assisted reproductive technology, more patients with infertility prefer to get pregnant by in vitro fertilization and embryo transplantation (IVF-ET). But the indications of IVF-ET must be strictly controlled by the clinicians. Case report: We described a case of a 29-year-old pregnant Chinese ...

  6. The role of management in an in vitro fertilization practice.

    Science.gov (United States)

    Masler, Steve; Strickland, Robert R

    2013-05-01

    An in vitro fertilization (IVF) practice is an enterprise. Like any enterprise, it has management that plays a major role, forming the structure, framework, and components that make the practice viable. Management of an IVF practice consists of two key teams: the fertility team and the management team. Management activities of the teams fall into eight core areas: business operations, financial, human resources, information technology, organizational governance, risk management, patient care systems, and quality management. Shady Grove Fertility Centers and Huntington Reproductive Center are two examples of professionally managed large fertility practices, one managed mostly centrally and the other largely managed in a decentralized way. Management is what takes a physician's IVF practice and converts it to a professional enterprise. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  7. Recent cadmium exposure among male partners may affect oocyte fertilization during in vitro fertilization (IVF)

    Science.gov (United States)

    Kim, Keewan; Fujimoto, Victor Y.; Parsons, Patrick J.; Steuerwald, Amy J.; Browne, Richard W.

    2010-01-01

    Purpose We recently reported evidence suggesting associations between urine cadmium concentrations, reflecting long-term exposure, measured in 25 female patients (relative risk = 1.41, P = 0.412) and 15 of their male partners (relative risk = 0.19, P = 0.097) and oocyte fertilization in vitro. Blood cadmium concentrations reflect more recent exposure. Methods We here incorporate those measures into our prior data set and employ multivariable log-binomial regression models to generate hypotheses concerning the relative effects of long-term and recent cadmium exposure on oocyte fertilization in vitro. Results No association is indicated for blood cadmium from women and oocyte fertilization, adjusted for urine cadmium and creatinine, blood lead and mercury, age, race/ethnicity and cigarette smoking (relative risk = 0.88, P = 0.828). However, we suggest an inverse adjusted association between blood cadmium from men and oocyte fertilization (relative risk = 0.66, P = 0.143). Conclusions These results suggest that consideration of long-term and recent exposures are both important for assessing the effect of partner cadmium levels on oocyte fertilization in vitro. PMID:20508982

  8. Psychological distress and in vitro fertilization outcome

    Science.gov (United States)

    Pasch, Lauri A.; Gregorich, Steven E.; Katz, Patricia K.; Millstein, Susan G.; Nachtigall, Robert D.; Bleil, Maria E.; Adler, Nancy E.

    2016-01-01

    Objective To examine whether psychological distress predicts IVF treatment outcome as well as whether IVF treatment outcome predicts subsequent psychological distress. Design Prospective cohort study over an 18-month period. Setting Five community and academic fertility practices. Patients Two hundred and two women who initiated their first IVF cycle. Interventions Women completed interviews and questionnaires at baseline and at 4, 10, and 18 months follow-up. Main Outcome Measures IVF cycle outcome and psychological distress. Results Using a binary logistic model including covariates (woman’s age, ethnicity, income, education, parity, duration of infertility, and time interval), pre-treatment depression and anxiety were not significant predictors of the outcome of the first IVF cycle. Using linear regression models including covariates (woman’s age, income, education, parity, duration of infertility, assessment point, time since last treatment cycle, and pre-IVF depression or anxiety), experiencing failed IVF was associated with higher post-IVF depression and anxiety. Conclusions IVF failure predicts subsequent psychological distress, but pre-IVF psychological distress does not predict IVF failure. Instead of focusing efforts on psychological interventions specifically aimed at improving the chance of pregnancy, these findings suggest that attention be paid to helping patients prepare for and cope with treatment and treatment failure. PMID:22698636

  9. Evaluation of Zona Pellucida Function for Sperm Penetration During In Vitro Fertilization in Pigs

    Science.gov (United States)

    TANIHARA, Fuminori; NAKAI, Michiko; KANEKO, Hiroyuki; NOGUCHI, Junko; OTOI, Takeshige; KIKUCHI, Kazuhiro

    2013-01-01

    Abstract In porcine oocytes, the function of the zona pellucida (ZP) with regard to sperm penetration or prevention of polyspermy is not well understood. In the present study, we investigated the effects of the ZP on sperm penetration during in vitro fertilization (IVF). We collected in vitro-matured oocytes with a first polar body (ZP+ oocytes). Some of them were freed from the ZP (ZP− oocytes) by two treatments (pronase and mechanical pipetting), and the effects of these treatments on sperm penetration parameters (sperm penetration rate and numbers of penetrated sperm per oocyte) were evaluated. There was no evident difference in the parameters between the two groups. Secondly, we compared the sperm penetration parameters of ZP+ and ZP− oocytes using frozen-thawed epididymal spermatozoa from four boars. Sperm penetration into ZP+ oocytes was found to be accelerated relative to ZP− oocytes. Thirdly, we evaluated the sperm penetration of ZP+ and ZP− oocytes at 1−10 h after IVF (3 h gamete co-incubation). The proportions of oocytes penetrated by sperm increased significantly with time in both groups; however, the number of penetrated sperm per oocyte did not increase in ZP− oocytes. Finally, we performed IVF using ZP− oocytes divided into control (3 h) and prolonged gamete co-incubation (5 h) groups. Greater numbers of sperm penetrated in the 5 h group than in the control group. These results suggest that the ZP and oolemma are not competent factors for prevention of polyspermy in our present porcine IVF system. However, it appears that ZP removal is one of the possibilities for reducing polyspermic penetration in vitro in pigs. PMID:23666494

  10. Utilization of third-party in vitro fertilization in the United States.

    Science.gov (United States)

    Kushnir, Vitaly A; Darmon, Sarah K; Shapiro, Alice J; Albertini, David F; Barad, David H; Gleicher, Norbert

    2017-03-01

    The use of in vitro fertilization that includes third-party in vitro fertilization is increasing. However, the relative contribution of third-party in vitro fertilization that includes the use of donor oocytes, sperm, or embryo and a gestational carrier to the birth cohort after in vitro fertilization is unknown. The purpose of this study was to examine the contribution of third-party in vitro fertilization to the in vitro fertilization birth cohort over the past decade. This retrospective analysis investigated 1,349,874 in vitro fertilization cycles that resulted in 421,525 live births and 549,367 liveborn infants in the United States from 2004-2013. Cycles were self-reported by fertility centers to a national registry: Society for Assisted Reproductive Technologies Clinic Outcome Reporting System. Third-party in vitro fertilization accounted for 217,030 (16.1%) of all in vitro fertilization cycles, 86,063 (20.4%) of all live births, and 115,024 (20.9%) of all liveborn infants. Overall, 39.7% of third-party in vitro fertilization cycles resulted in a live birth, compared with 29.6% of autologous in vitro fertilization cycles. Use of third-party in vitro fertilization increased with maternal age and accounted for 42.2% of all in vitro fertilization cycles and 75.3% of all liveborn infants among women >40 years old. Oocyte donation was the most common third-party in vitro fertilization technique, followed by sperm donation. Over the study period, annual cycle volume and live birth rates gradually increased for both autologous in vitro fertilization and third-party in vitro fertilization (Pvitro fertilization modalities were used, followed by oocyte donation. Third-party in vitro fertilization use and efficacy have increased over the past decade, now comprising >20% of the total in vitro fertilization birth cohort. In women who are >40 years old, third-party in vitro fertilization has become the dominant treatment. Copyright © 2016 Elsevier Inc

  11. Optimizing Inflammatory Bowel Disease for Successful In Vitro Fertilization.

    Science.gov (United States)

    Pan, Jason J; Cannon, Mora; Kane, Edward; Konijeti, Gauree

    2016-08-01

    We present a nulliparous woman with mild to moderate ulcerative colitis (UC) and multiple failed cycles of in vitro fertilization (IVF) in whom we achieved a successful, viable pregnancy following clinical and endoscopic UC remission. Infertile patients with inflammatory bowel disease who have failed multiple cycles of IVF should try to achieve clinical remission and mucosal healing (absence of erosions or ulcers) prior to reattempting conception. Furthermore, deficiencies in vitamin B12, vitamin D, and iron should be addressed.

  12. In vitro fertilization/intracytoplasmic sperm injection for male infertility

    OpenAIRE

    Rubina Merchant; Goral Gandhi; Allahbadia, Gautam N.

    2011-01-01

    Progress in the field of assisted reproduction, and particularly micromanipulation, now heralds a new era in the management of severe male factor infertility, not amenable to medical or surgical correction. By overcoming natural barriers to conception, in vitro fertilization and embryo transfer (IVF-ET), subzonal sperm insemination, partial zona dissection, and intracytoplasmatic injection of sperm (ICSI) now offer couples considered irreversibly infertile, the option of parenting a genetical...

  13. Effects of extracellular potassium concentrations on acrosome reaction and polyspermy during in vitro fertilization and subsequent development in vitro in the pig.

    Science.gov (United States)

    Lim, J G; Kim, N H; Lee, H T; Chung, K S

    1997-10-01

    Potassium (K(+)) concentration in the mammalian oviduct and uterus is particularly interesting due to its unusually high concentration (12 to 25 mM) compared with that in the blood stream (3 to 6 mM). In this study we examined the effects of various K(+) concentrations in the fertilization medium on polyspermy and subsequent in vitro development of porcine oocytes. In the absence of K(+) in the fertilization medium, sperm penetration was not observed. The incidence of polyspermy was significantly higher in the fertilization medium that contained 6 or 12 mM K(+) as compared with 3 mM K(+). The mean number of sperm penetrated in oocytes in medium with 6 and 12 mM K(+) was higher than in medium with 3 mM K(+). The addition of 3, 6 or 12 mM K(+) to the fertilization medium did not significantly affect the proportion of zygotes that developed to the blastocyst stage (14.1, 12.4 and 15.0%, respectively). Chlorotetracycline (CTC) analysis was used to determine the capacitation and acrosome reaction of spermatozoa incubated for 3 h with various concentrations of K(+). The number of acrosome reacted spermatozoa decreased with increasing K(+) concentration. These results suggest that extracellular K(+) in the fertilization medium affects sperm acrosome reaction which may be related to the sperm penetration.

  14. Effect of punicalagin on proliferation of porcine ovarian granulosa cells in vitro

    OpenAIRE

    Dagmara Packová; Adriana Kolesárová

    2016-01-01

    Punicalagin is a major component responsible for pomegranate's (Punica granatum) antioxidant properties. Punicalagin is the predominant ellagitannin of Punica granatum and present in two isomeric forms: punicalagin α and β. Punicalagin is metabolised to ellagic acid (antioxidant) and microorganisms present in colon can metabolize ellagic acid to urolithins. The aim of in vitro study was to examine the effect of punicalagin on mitochondrial activity and markers of proliferation in porcine ovar...

  15. Enhanced delivery of ketobemidone through porcine buccal mucosa in vitro via more lipophilic ester prodrugs

    DEFF Research Database (Denmark)

    Hansen, L.B.; Christrup, Lona Louring; Bundgaard, H.

    1992-01-01

    The in vitro penetration of ketobemidone and various ester prodrugs through porcine buccal mucosa in a modified Ussing chamber was investigated in order to support the selection of a prodrug derivative with optimal buccal absorption. The nine esters studied included carboxylic acid and carbonate...... a plateau or a slight decrease occurred. No toxic effects of ketobemidone or the prodrugs on the buccal membrane were observed as judged from monitoring of the electrical properties of the membrane....

  16. Effect of coculturing spermatozoa with oviductal cells on the incidence of polyspermy in pig in vitro fertilization.

    Science.gov (United States)

    Dubuc, A; Sirard, M A

    1995-07-01

    It is known that oviductal cells play an important role in fertilization in vivo. We were interested in the effect of those cells on spermatozoa and their influence on the incidence of polyspermy in pig in vitro fertilization (IVF) when cocultured with spermatozoa. Oviductal cells are believed to select a highly fecund population of spermatozoa. By coculturing spermatozoa with oviductal cells it is possible to reduce the number of spermatozoa confronting the eggs at fertilization, reducing the incidence of polyspermy. In our study, spermatozoa were cocultured with oviductal cells for 30 min so that they could bind to the oviductal cells. Both bound and unbound spermatozoa were used for fertilization. The results show that the spermatozoa that were bound to the oviductal cells were capable of fertilizing the eggs, but the nonbound spermatozoa had a reduced penetration incidence. With the bound sperm, polyspermy incidence decreased and the two-pronuclei proportion increased. We also found that with time the spermatozoa released from the cells had better motility than those that did not bind. Therefore, it is our belief that oviductal cells can be used for porcine IVF resulting in a lower polyspermy incidence and higher pronuclei incidence.

  17. Low concentrations of MEM vitamins during in vitro maturation of porcine oocytes improves subsequent parthenogenetic development.

    Science.gov (United States)

    Naruse, Kenji; Kim, Hong Rye; Shin, Young Min; Chang, Suk Min; Lee, Hye Ran; Park, Chang Sik; Jin, Dong Il

    2007-01-15

    To investigate the effects of water-soluble vitamin supplementation for IVM/IVC of porcine oocytes and evaluate maturation and developmental capacity in vitro, porcine cumulus oocyte complexes (COCs) was matured in NCSU-23-based medium with water-soluble vitamins for 44 h and then cultured in PZM-3 for 7 days following activation. The COCs were allocated into five treatment groups and matured in various concentrations of MEM vitamins (control, 0.05, 0.1, 0.2, 0.4, and 1x). Metaphase II plates of the cumulus-free oocytes were observed following Hoechest 33258 staining. The COCs were allocated into four treatment groups, matured in various concentrations of MEM vitamins (control, 0.05, 0.1, 0.2, and 0.4x) and cultured in PZM-3 following activation. Also, COCS were matured without MEM vitamins and cultured in PZM-3 with various concentrations (control, 0.1, 0.4, 1.0, and 2.0 x) of MEM vitamins. Furthermore, 2 x 2 factorial (IVM/IVC) experiments were performed in IVM medium with or without 0.05 x MEM vitamins and IVC medium with or without 0.4x MEM vitamins to examine the in vitro development of parthenogenetic embryos. Maturation rates of COCs treated with MEM vitamins did not differ significantly among groups. However, compared to the control group, oocytes matured with the addition of 0.05 x MEM vitamins developed to blastocysts at a higher percentage (PMEM vitamins. Total cell number of blastocysts was significantly higher in the 0.05 x group. Addition of 0.4x MEM vitamins decreased (PMEM vitamins-treated group. In contrast, addition of vitamins to PZM-3 medium for in vitro culture of activated porcine oocytes did not affect development. In conclusion, addition of a low concentration of MEM vitamins to IVM medium for porcine oocytes enhanced subsequent development and improved embryo quality.

  18. Calcium-sensing receptor (CASR) is involved in porcine in vitro fertilisation and early embryo development.

    Science.gov (United States)

    Liu, C; Liu, Y; Larsen, K; Hou, Y P; Callesen, H

    2017-07-17

    It has been demonstrated that extracellular calcium is necessary in fertilisation and embryo development but the mechanism is still not well understood. The present study mainly focussed on the extracellular calcium effector called the calcium-sensing receptor (CASR) and examined its expression in porcine gametes and embryos and its function during fertilisation and early embryo development. By using reverse transcription polymerase chain reaction, CASR was found to be expressed in porcine oocytes, spermatozoa and embryos at different developmental stages. Functionally, medium supplementation with a CASR agonist or an antagonist during in vitro fertilisation (IVF) and in vitro culture (IVC) was tested. During fertilisation, the presence of a CASR agonist increased sperm penetration rate and decreased polyspermy rate leading to an increased normal fertilisation rate. During embryo development, for the IVF embryos, agonist treatment during IVC significantly increased cleavage rate and blastocyst formation rate compared with the control group. Furthermore, parthenogenetically activated embryos showed similar results with lower cleavage and blastocyst formation rates in the antagonist group than in the other groups. It was concluded that CASR, as the effector of extracellular calcium, modulates porcine fertilisation and early embryo development.

  19. Hair mercury concentrations and in vitro fertilization (IVF) outcomes among women from a fertility clinic

    Science.gov (United States)

    Ehrlich, Shelley; Smith, Kristen; Williams, Paige L.; Chavarro, Jorge E.; Batsis, Maria; Toth, Thomas L.; Hauser, Russ

    2015-01-01

    Total hair mercury (Hg) was measured among 205 women undergoing in vitro fertilization (IVF) treatment and the association with prospectively collected IVF outcomes (229 IVF cycles) was evaluated. Hair Hg levels (median=0.62 ppm, range: 0.03-5.66 ppm) correlated with fish intake (r=0.59), and exceeded the recommended EPA reference of 1ppm in 33% of women. Generalized linear mixed models with random intercepts accounting for within-woman correlations across treatment cycles were used to evaluate the association of hair Hg with IVF outcomes adjusted for age, body mass index, race, smoking status, infertility diagnosis, and protocol type. Hair Hg levels were not related to ovarian stimulation outcomes (peak estradiol levels, total and mature oocyte yields) or to fertilization rate, embryo quality, clinical pregnancy rate or live birth rate. PMID:25601638

  20. RNA profiles of porcine embryos during genome activation reveal complex metabolic switch sensitive to in vitro conditions.

    Directory of Open Access Journals (Sweden)

    Olga Østrup

    Full Text Available Fertilization is followed by complex changes in cytoplasmic composition and extensive chromatin reprogramming which results in the abundant activation of totipotent embryonic genome at embryonic genome activation (EGA. While chromatin reprogramming has been widely studied in several species, only a handful of reports characterize changing transcriptome profiles and resulting metabolic changes in cleavage stage embryos. The aims of the current study were to investigate RNA profiles of in vivo developed (ivv and in vitro produced (ivt porcine embryos before (2-cell stage and after (late 4-cell stage EGA and determine major metabolic changes that regulate totipotency. The period before EGA was dominated by transcripts responsible for cell cycle regulation, mitosis, RNA translation and processing (including ribosomal machinery, protein catabolism, and chromatin remodelling. Following EGA an increase in the abundance of transcripts involved in transcription, translation, DNA metabolism, histone and chromatin modification, as well as protein catabolism was detected. The further analysis of members of overlapping GO terms revealed that despite that comparable cellular processes are taking place before and after EGA (RNA splicing, protein catabolism, different metabolic pathways are involved. This strongly suggests that a complex metabolic switch accompanies EGA. In vitro conditions significantly altered RNA profiles before EGA, and the character of these changes indicates that they originate from oocyte and are imposed either before oocyte aspiration or during in vitro maturation. IVT embryos have altered content of apoptotic factors, cell cycle regulation factors and spindle components, and transcription factors, which all may contribute to reduced developmental competence of embryos produced in vitro. Overall, our data are in good accordance with previously published, genome-wide profiling data in other species. Moreover, comparison with mouse and

  1. Treatment of porcine oocytes with MEM vitamins during in vitro maturation improves subsequent blastocyst development following nuclear transfer.

    Science.gov (United States)

    Naruse, Kenji; Quan, Yan-Shi; Choi, Su-Min; Park, Chang-Sik; Jin, Dong-Il

    2007-06-01

    This study was carried out to investigate the effects of minimum essential medium (MEM) vitamins during in vitro maturation (IVM)/in vitro culture (IVC) of porcine nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. Porcine cumulus-oocyte complexes (COCs) were divided into five groups, matured for 44 h in maturation medium with various concentrations of MEM vitamins (0, 0.05, 0.1, 0.2 and 0.4%), and observed for maturation rate. Also, COCs were matured in NUSU-23 media without MEM vitamins for 44 h and cultured in PZM-3 media with various concentrations of MEM vitamins (0, 0.05, 0.4 and 1.0%) for 6 days following nuclear transfer. Factorial (IVM/IVC) experiments were also performed in NCSU-23 medium with or without 0.05% MEM vitamins and PZM-3 medium with or without 0.4% MEM vitamins. They were then tested by examining in vitro development of the porcine reconstructed embryos. The maturation rates of the COCs treated with the MEM vitamins did not differ significantly among the MEM vitamin-treated groups. Addition of vitamins to culture medium did not affect development of porcine reconstructed embryos in vitro. However, addition of low concentrations of MEM vitamins only to maturation medium increased (PMEM vitamins to IVC medium did not enhance the developmental rate compared with the control group. Thus, addition of MEM vitamins to IVM medium could improve subsequent blastocyst development of porcine NT embryos.

  2. Quality management systems for your in vitro fertilization clinic′s laboratory: Why bother?

    National Research Council Canada - National Science Library

    Jan I Olofsson; Manish R Banker; Late Peter Sjoblom

    2013-01-01

    Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF...

  3. Quality management systems for your in vitro fertilization clinic's laboratory: Why bother?

    National Research Council Canada - National Science Library

    Olofsson, Jan I; Banker, Manish R; Sjoblom, Late Peter

    2013-01-01

    Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF...

  4. Ochratoxin A promotes porcine circovirus type 2 replication in vitro and in vivo.

    Science.gov (United States)

    Gan, Fang; Zhang, Zheqian; Hu, Zhihua; Hesketh, John; Xue, Hongxia; Chen, Xingxiang; Hao, Shu; Huang, Yu; Cole Ezea, Patience; Parveen, Fahmida; Huang, Kehe

    2015-03-01

    Ochratoxin A (OTA), a worldwide mycotoxin found in food and feeds, is a potent nephrotoxin in animals and humans. Porcine circovirus-associated disease (PCVAD), including porcine dermatitis and nephropathy syndrome, is a worldwide swine disease. To date, little is known concerning the relationship between OTA and porcine circovirus type 2 (PCV2), the primary causative agent of PCVAD. The effects of OTA on PCV2 replication and their mechanisms were investigated in vitro and in vivo. The results in vitro showed that low doses of OTA significantly increased PCV2 DNA copies and the number of infected cells. Maximum effects were observed at 0.05 μg/ml OTA. The results in vivo showed that PCV2 replication was significantly increased in serum and tissues of pigs fed 75 μg/kg OTA compared with the control group and pigs fed 150 μg/kg OTA. In addition, low doses of OTA significantly depleted reduced glutathione and mRNA expression of NF-E2-related factor 2 and γ-glutamylcysteine synthetase; increased reactive oxygen species, oxidants, and malondialdehyde; and induced p38 and ERK1/2 phosphorylation in PK15 cells. Adding N-acetyl-L-cysteine reversed the changes induced by OTA. Knockdown of p38 and ERK1/2 by their respective specific siRNAs or inhibition of p38 and ERK1/2 phosphorylation by their respective inhibitors (SB203580 and U0126) eliminated the increase in PCV2 replication induced by OTA. These data indicate that low doses of OTA promoted PCV2 replication in vitro and in vivo via the oxidative stress-mediated p38/ERK1/2 MAPK signaling pathway. This suggests that low doses of OTA are potentially harmful to animals, as they enhance virus replication, and partly explains why the morbidity and severity of PCVAD vary significantly in different pig farms. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. 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.

  6. In Vitro and In Vivo Study of a Novel Porcine Collagen Membrane for Guided Bone Regeneration

    Directory of Open Access Journals (Sweden)

    Eisner Salamanca

    2016-11-01

    Full Text Available For years, in order to improve bone regeneration and prevent the need of a second stage surgery to remove non-resorbable membranes, biological absorbable membranes have gradually been developed and applied in guided tissue regeneration (GTR. The present study’s main objective was to achieve space maintenance and bone regeneration using a new freeze-dried developed porcine collagen membrane, and compare it with an already commercial collagen membrane, when both were used with a bovine xenograft in prepared alveolar ridge bone defects. Prior to surgery, the membrane’s vitality analysis showed statistically significant higher cell proliferation in the test membrane over the commercial one. In six beagle dogs, commercial bone xenograft was packed in lateral ridge bone defects prepared in the left and right side and then covered with test porcine collagen membrane or commercial collagen membrane. Alveolar height changes were measured. Histomorphometric results, in vitro and in vivo properties indicated that the new porcine collagen membrane is biocompatible, enhances bone xenograft osteoconduction, and reduces the alveolar ridge height reabsorption rate.

  7. Sexual function and fertility quality of life in women using in vitro fertilization.

    Science.gov (United States)

    Smith, Nicole K; Madeira, Jody; Millard, Heather R

    2015-04-01

    Women undergoing fertility treatment are likely to report negative changes in sexual function. With the rapid increase in the number of women pursuing in vitro fertilization (IVF), there is a need to better understand how IVF impacts a woman's sexual experiences and quality of life. This study has three purposes: (i) test the psychometric properties of the Sexual Functioning Questionnaire (SFQ) in a sample of U.S. women undergoing IVF; (ii) compare sexual function of women undergoing IVF with a nonclinical sample of U.S. women; and (iii) identify the aspects of sexual function most related to fertility quality of life (FertiQoL). A total of 136 women who had recently undergone or who were currently undergoing IVF completed a web-based, cross-sectional survey about sexual experiences and quality of life. Data were collected on infertility diagnosis, length of infertility, number of IVF cycles, pregnancy, and birth outcomes. Six domains of sexual function as well as the medical impact of IVF were assessed using the SFQ. A validated instrument was used to measure FertiQoL. Reliability analyses for the SFQ indicate sufficiently strong fit (Cronbach's alpha = 0.79 to 0.89). Compared with a nonclinical sample, women undergoing IVF scored significantly lower in sexual interest, desire, orgasm, satisfaction, sexual activity, and overall sexual function (P Women undergoing IVF may be at particular risk for sexual problems. Sexual function issues may markedly impact overall quality of life during fertility treatment and should be addressed as an important component of comprehensive care. © 2015 International Society for Sexual Medicine.

  8. Oxidative stress induced by zearalenone in porcine granulosa cells and its rescue by curcumin in vitro.

    Directory of Open Access Journals (Sweden)

    Xunsi Qin

    Full Text Available Oxidative stress (OS, as a signal of aberrant intracellular mechanisms, plays key roles in maintaining homeostasis for organisms. The occurrence of OS due to the disorder of normal cellular redox balance indicates the overproduction of reactive oxygen species (ROS and/or deficiency of antioxidants. Once the balance is broken down, repression of oxidative stress is one of the most effective ways to alleviate it. Ongoing studies provide remarkable evidence that oxidative stress is involved in reproductive toxicity induced by various stimuli, such as environmental toxicants and food toxicity. Zearalenone (ZEA, as a toxic compound existing in contaminated food products, is found to induce mycotoxicosis that has a significant impact on the reproduction of domestic animals, especially pigs. However, there is no information about how ROS and oxidative stress is involved in the influence of ZEA on porcine granulosa cells, or whether the stress can be rescued by curcumin. In this study, ZEA-induced effect on porcine granulosa cells was investigated at low concentrations (15 μM, 30 μM and 60 μM. In vitro ROS levels, the mRNA level and activity of superoxide dismutase, glutathione peroxidase and catalase were obtained. The results showed that in comparison with negative control, ZEA increased oxidative stress with higher ROS levels, reduced the expression and activity of antioxidative enzymes, increased the intensity of fluorogenic probes 2', 7'-Dichlorodihydrofluorescin diacetate and dihydroethidium in flow cytometry assay and fluorescence microscopy. Meanwhile, the activity of glutathione (GSH did not change obviously following 60 μM ZEA treatment. Furthermore, the underlying protective mechanisms of curcumin on the ZEA-treated porcine granulosa cells were investigated. The data revealed that curcumin pre-treatment significantly suppressed ZEA-induced oxidative stress. Collectively, porcine granulosa cells were sensitive to ZEA, which may induce

  9. In vitro fertilization/intracytoplasmic sperm injection for male infertility

    Directory of Open Access Journals (Sweden)

    Rubina Merchant

    2011-01-01

    Full Text Available Progress in the field of assisted reproduction, and particularly micromanipulation, now heralds a new era in the management of severe male factor infertility, not amenable to medical or surgical correction. By overcoming natural barriers to conception, in vitro fertilization and embryo transfer (IVF-ET, subzonal sperm insemination, partial zona dissection, and intracytoplasmatic injection of sperm (ICSI now offer couples considered irreversibly infertile, the option of parenting a genetically related child. However, unlike IVF, which necessitates an optimal sperm number and function to successfully complete the sequence of events leading to fertilization, micromanipulation techniques, such as ICSI, involving the direct injection of a spermatozoon into the oocyte, obviate all these requirements and may be used to alleviate severe male factor infertility due to the lack of sperm in the ejaculate due to severely impaired spermatogenesis (non-obstructive azoospermia or non-reconstructable reproductive tract obstruction (obstructive azoospermia. ICSI may be performed with fresh or cryopreserved ejaculate sperm where available, microsurgically extracted epididymal or testicular sperm with satisfactory fertilization, clinical pregnancy, and ongoing pregnancy rates. However, despite a lack of consensus regarding the genetic implications of ICSI or the application and efficacy of preimplantation genetic diagnosis prior to assisted reproductive technology (ART, the widespread use of ICSI, increasing evidence of the involvement of genetic factors in male infertility and the potential risk of transmission of genetic disorders to the offspring, generate major concerns with regard to the safety of the technique, necessitating a thorough genetic evaluation of the couple, classification of infertility and adequate counseling of the implications and associated risks prior to embarking on the procedure. The objective of this review is to highlight the indications

  10. In vitro fertilization/intracytoplasmic sperm injection for male infertility.

    Science.gov (United States)

    Merchant, Rubina; Gandhi, Goral; Allahbadia, Gautam N

    2011-01-01

    Progress in the field of assisted reproduction, and particularly micromanipulation, now heralds a new era in the management of severe male factor infertility, not amenable to medical or surgical correction. By overcoming natural barriers to conception, in vitro fertilization and embryo transfer (IVF-ET), subzonal sperm insemination, partial zona dissection, and intracytoplasmatic injection of sperm (ICSI) now offer couples considered irreversibly infertile, the option of parenting a genetically related child. However, unlike IVF, which necessitates an optimal sperm number and function to successfully complete the sequence of events leading to fertilization, micromanipulation techniques, such as ICSI, involving the direct injection of a spermatozoon into the oocyte, obviate all these requirements and may be used to alleviate severe male factor infertility due to the lack of sperm in the ejaculate due to severely impaired spermatogenesis (non-obstructive azoospermia) or non-reconstructable reproductive tract obstruction (obstructive azoospermia). ICSI may be performed with fresh or cryopreserved ejaculate sperm where available, microsurgically extracted epididymal or testicular sperm with satisfactory fertilization, clinical pregnancy, and ongoing pregnancy rates. However, despite a lack of consensus regarding the genetic implications of ICSI or the application and efficacy of preimplantation genetic diagnosis prior to assisted reproductive technology (ART), the widespread use of ICSI, increasing evidence of the involvement of genetic factors in male infertility and the potential risk of transmission of genetic disorders to the offspring, generate major concerns with regard to the safety of the technique, necessitating a thorough genetic evaluation of the couple, classification of infertility and adequate counseling of the implications and associated risks prior to embarking on the procedure. The objective of this review is to highlight the indications, advantages

  11. Inositol and In Vitro Fertilization with Embryo Transfer

    Directory of Open Access Journals (Sweden)

    G. Simi

    2017-01-01

    Full Text Available Recently, studies on inositol supplementation during in vitro fertilization program (IVF have gained particular importance due to the effect of this molecule on reducing insulin resistance improving ovarian function, oocyte quality, and embryo and pregnancy rates and reducing gonadotropin amount during stimulation. Inositol and its isoforms, especially myoinositol (MYO, are often used as prestimulation therapy in infertile patients undergoing IVF cycle. Inositol supplementation started three months before ovarian stimulation, resulting in significant improvements in hormonal responses, reducing the amount of FSH necessary for optimal follicle development and serum levels of 17beta-estradiol measured the day of hCG injection. As shown by growing number of trials, MYO supplementation improves oocyte quality by reducing the number of degenerated and immature oocytes, in this way increasing the quality of embryos produced. Inositol can also improve the quality of sperm parameters in those patients affected by oligoasthenoteratozoospermia.

  12. Social and ethical aspects of in vitro fertilization.

    Science.gov (United States)

    Heitman, E

    1999-01-01

    In vitro fertilization (IVF) stands out as one of the contemporary period's most extraordinary technologies, and its social and ethical consequences among the most far reaching. Despite its uncertain effectiveness and medical consequences, IVF has contributed significantly to the medicalization of infertility and the increasingly imperative character of reproductive technology. New developments in IVF, particularly oocyte donation, have created new definitions of treatable infertility and new social needs for IVF; when the technology does not result in pregnancy or healthy babies, these developments have created profound new disappointments. IVF and the commodification of the extracorporeal embryo have also confused the social meaning and legal definition of parenthood. Ultimately the relationship between prospective parents, infertility specialists, and the embryos that they create is a highly ambiguous one. This ambiguity is likely to be a long-term characteristic of efforts to develop, use, and assess assisted reproductive technologies.

  13. The Role of Ultrasonography in In-vitro Fertilization And Embryo ...

    African Journals Online (AJOL)

    CONCLUSION: Ultrasonography is now the single most important instrument in in-vitro fertilization programmes and gynaecologists with interest in reproductive medicine need necessarily to obtain a formal training in its use. KEY WORDS: ultrasonography, in-vitro fertilization, infertility, assisted reproduction technology ...

  14. Dropout rates in couples undergoing in vitro fertilization and intrauterine insemination

    NARCIS (Netherlands)

    Bensdorp, Alexandra J.; Tjon-Kon-Fat, Raissa; Verhoeve, Harold; Koks, Carolien; Hompes, Peter; Hoek, Annemieke; de Bruin, Jan Peter; Cohlen, Ben; Hoozemans, Diederik; Broekmans, Frank; van Bomme, Peter; Smeenk, Jesper; Mol, Ben W. J.; van der Veen, Fulco; van Wely, Madelon

    2016-01-01

    Objective: To compare dropout rates in couples undergoing conventional in vitro fertilization with single embryo transfer (IVF-SET), in vitro fertilization in a modified natural cycle (IVF-MNC) or intrauterine insemination with ovarian stimulation (IUI-OS). Study design: Secondary analysis of a

  15. Dropout rates in couples undergoing in vitro fertilization and intrauterine insemination

    NARCIS (Netherlands)

    Bensdorp, Alexandra J.; Tjon-Kon-Fat, Raissa; Verhoeve, Harold; Koks, Carolien; Hompes, Peter; Hoek, Annemieke; de Bruin, Jan Peter; Cohlen, Ben; Hoozemans, Diederik; Broekmans, Frank; van Bomme, Peter; Smeenk, Jesper; Mol, Ben W J; van der Veen, Fulco; van Wely, Madelon

    2016-01-01

    Objective To compare dropout rates in couples undergoing conventional in vitro fertilization with single embryo transfer (IVF-SET), in vitro fertilization in a modified natural cycle (IVF-MNC) or intrauterine insemination with ovarian stimulation (IUI-OS). Study design Secondary analysis of a

  16. Porcine ear skin as a biological substrate for in vitro testing of sunscreen performance.

    Science.gov (United States)

    Sohn, Myriam; Korn, Verena; Imanidis, Georgios

    2015-01-01

    The purpose of the study was to examine the use of skin from porcine ears as a biological substrate for in vitro testing of sunscreens in order to overcome the shortcomings of the presently used polymethylmethacrylate (PMMA) plates that generally fail to yield a satisfactory correlation between sun protection factors (SPF) in vitro and in vivo. Trypsin-separated stratum corneum and heat-separated epidermis provided UV-transparent substrates that were laid on quartz or on PMMA plates. These were used to determine surface roughness by chromatic confocal imaging and to measure SPF in vitro of 2 sunscreens by diffuse transmission spectroscopy. The recovered skin layers showed a lower roughness than full-thickness skin but yielded SPF in vitro values that more accurately reflected the SPF determined in vivo by a validated procedure than PMMA plates, although the latter had in part roughness values identical to those of intact skin. Combination of skin tissue with a high roughness PMMA plate also provided accurate SPF in vitro. Besides roughness, the improved affinity of the sunscreen to the skin substrate compared to PMMA plates may explain the better in vitro prediction of SPF achieved with the use of a biological substrate. © 2014 S. Karger AG, Basel.

  17. Thiouracil-Forming Bacteria Identified and Characterized upon Porcine In Vitro Digestion of Brassicaceae Feed

    Science.gov (United States)

    Kiebooms, Julie A. L.; Wauters, Jella; Vanden Bussche, Julie; Houf, Kurt; De Vos, Paul; Van Trappen, Stefanie; Cleenwerck, Ilse

    2014-01-01

    In recent years, the frequent detection of the banned thyreostat thiouracil (TU) in livestock urine has been related to endogenous TU formation following digestion of glucosinolate-rich Brassicaceae crops. Recently, it was demonstrated that, upon in vitro digestion of Brassicaceae, fecal bacteria induce TU detection in livestock (porcine livestock > bovines). Therefore, the present study was intended to isolate and identify bacteria involved in this intestinal TU formation upon Brassicaceae digestion and to gain more insight into the underlying mechanism in porcine livestock. Twenty porcine fecal inocula (gilts and multiparous sows) were assessed through static in vitro colonic-digestion simulations with rapeseed. After derivatization and extraction of the fecal suspensions, TU was analyzed using liquid chromatography-tandem mass spectrometry (LC-MS2). On average, lower TU concentrations were observed in fecal colonic simulations in gilts (8.35 ng g−1 rapeseed ± 3.42 [mean ± standard deviation]) than in multiparous sows (52.63 ng g−1 ± 16.17), which correlates with maturation of the gut microbial population with age. Further exploration of the mechanism showed cell-dependent activity of the microbial conversion and sustained TU-forming activity after subjection of the fecal inoculum to moderate heat over a time span of up to 30 min. Finally, nine TU-producing bacterial species were successfully isolated and identified by a combination of biochemical and molecular techniques as Escherichia coli (n = 5), Lactobacillus reuteri (n = 2), Enterococcus faecium (n = 1), and Salmonella enterica subsp. arizonae (n = 1). This report demonstrates that endogenous formation of TU is Brassicaceae induced and occurs under colonic conditions most likely through myrosinase-like enzyme activity expressed by different common intestinal bacterial species. PMID:25261511

  18. EFFECT OF QUERCETIN AND T-2 TOXIN ON ANTIOXIDANT PARAMETERS OF PORCINE BLOOD IN VITRO

    Directory of Open Access Journals (Sweden)

    Peter Petruška

    2012-10-01

    Full Text Available T-2 toxin, a trichothecene mycotoxin, is considered to be a one of the most toxic compounds that is produced by molds, particularly the Fusarium species. Mycotoxins can contaminate a large variety of feed mixtures, and could cause serious health problems to domestic livestock and humans when consumed. The aim of the present study was to investigate the effect of quercetin and T-2 toxin on some antioxidants parameters (superoxide dismutase - SOD, glutathione peroxidase - GPx in porcine blood in vitro. Application of quercetin in different doses (1, 10, 100 mg.ml-1 and T-2 toxin (1000 ng.ml-1 has caused significant decrease of SOD activity. Differences among the groups in the case of GPx activity remained insignificant (P<0.05. In this study the additions of quercetin and T-2 toxin in different doses in porcine blood in vitro caused significantly (P<0.05 lower SOD activity in all experimental groups in comparison with the control group.

  19. Expression of nucleolar-related proteins in porcine preimplantation embryos produced in vivo and in vitro

    DEFF Research Database (Denmark)

    Bjerregaard, Bolette; Wrenzycki, Christine; Strejcek, Frantisek

    2004-01-01

    embryonic mRNA transcription. Localization of proteins involved in the rRNA transcription (upstream binding factor [UBF], topoisomerase I, RNA polymerase I [RNA Pol I], and the RNA Pol I-associated factor PAF53) and processing (fibrillarin, nucleophosmin, and nucleolin) was assessed by immunocytochemistry....... Moreover, immunolocalization of RNA Pol I, but not of UBF, and the mRNA expression of PAF53 and UBF were significantly reduced or absent after culture with alpha-amanitin, indicating that RNA Pol I, PAF53, and presumably, UBF are derived from de novo embryonic transcription. Embryonic genomic activation...... was delayed in porcine embryos produced in vitro compared to the in vivo-derived counterparts with respect to mRNAs encoding PAF53 and UBF. Moreover, differences existed in the mRNA expression patterns of pRb between in vivo- and in vitro-developed embryos. These findings show, to our knowledge for the first...

  20. Karyotype characterization of in vivo- and in vitro-derived porcine parthenogenetic cell lines.

    Science.gov (United States)

    Liu, Qiang; Zhang, Manling; Hou, Dongxia; Han, Xuejie; Jin, Yong; Zhao, Lihua; Nie, Xiaowei; Zhou, Xin; Yun, Ting; Zhao, Yuhang; Huang, Xianghua; Hou, Daorong; Yang, Ning; Wu, Zhaoqiang; Li, Xueling; Li, Rongfeng

    2014-01-01

    Mammalian haploid cell lines provide useful tools for both genetic studies and transgenic animal production. To derive porcine haploid cells, three sets of experiments were conducted. First, genomes of blastomeres from 8-cell to 16-cell porcine parthenogenetically activated (PA) embryos were examined by chromosome spread analysis. An intact haploid genome was maintained by 48.15% of blastomeres. Based on this result, two major approaches for amplifying the haploid cell population were tested. First, embryonic stem-like (ES-like) cells were cultured from PA blastocyst stage embryos, and second, fetal fibroblasts from implanted day 30 PA fetuses were cultured. A total of six ES-like cell lines were derived from PA blastocysts. No chromosome spread with exactly 19 chromosomes (the normal haploid complement) was found. Four cell lines showed a tendency to develop to polyploidy (more than 38 chromosomes). The karyotypes of the fetal fibroblasts showed different abnormalities. Cells with 19-38 chromosomes were the predominant karyotype (59.48-60.91%). The diploid cells were the second most observed karyotype (16.17%-22.73%). Although a low percentage (3.45-8.33%) of cells with 19 chromosomes were detected in 18.52% of the fetus-derived cell lines, these cells were not authentic haploid cells since they exhibited random losses or gains of some chromosomes. The haploid fibroblasts were not efficiently enriched via flow cytometry sorting. On the contrary, the diploid cells were efficiently enriched. The enriched parthenogenetic diploid cells showed normal karyotypes and expressed paternally imprinted genes at extremely low levels. We concluded that only a limited number of authentic haploid cells could be obtained from porcine cleavage-stage parthenogenetic embryos. Unlike mouse, the karyotype of porcine PA embryo-derived haploid cells is not stable, long-term culture of parthenogenetic embryos, either in vivo or in vitro, resulted in abnormal karyotypes. The porcine PA

  1. Karyotype characterization of in vivo- and in vitro-derived porcine parthenogenetic cell lines.

    Directory of Open Access Journals (Sweden)

    Qiang Liu

    Full Text Available Mammalian haploid cell lines provide useful tools for both genetic studies and transgenic animal production. To derive porcine haploid cells, three sets of experiments were conducted. First, genomes of blastomeres from 8-cell to 16-cell porcine parthenogenetically activated (PA embryos were examined by chromosome spread analysis. An intact haploid genome was maintained by 48.15% of blastomeres. Based on this result, two major approaches for amplifying the haploid cell population were tested. First, embryonic stem-like (ES-like cells were cultured from PA blastocyst stage embryos, and second, fetal fibroblasts from implanted day 30 PA fetuses were cultured. A total of six ES-like cell lines were derived from PA blastocysts. No chromosome spread with exactly 19 chromosomes (the normal haploid complement was found. Four cell lines showed a tendency to develop to polyploidy (more than 38 chromosomes. The karyotypes of the fetal fibroblasts showed different abnormalities. Cells with 19-38 chromosomes were the predominant karyotype (59.48-60.91%. The diploid cells were the second most observed karyotype (16.17%-22.73%. Although a low percentage (3.45-8.33% of cells with 19 chromosomes were detected in 18.52% of the fetus-derived cell lines, these cells were not authentic haploid cells since they exhibited random losses or gains of some chromosomes. The haploid fibroblasts were not efficiently enriched via flow cytometry sorting. On the contrary, the diploid cells were efficiently enriched. The enriched parthenogenetic diploid cells showed normal karyotypes and expressed paternally imprinted genes at extremely low levels. We concluded that only a limited number of authentic haploid cells could be obtained from porcine cleavage-stage parthenogenetic embryos. Unlike mouse, the karyotype of porcine PA embryo-derived haploid cells is not stable, long-term culture of parthenogenetic embryos, either in vivo or in vitro, resulted in abnormal karyotypes. The

  2. Differential expression of key subunits of SWI/SNF chromatin remodeling complexes in porcine embryos derived in vitro or in vivo.

    Science.gov (United States)

    Cabot, Birgit; Tseng, Yu-Chun; Crodian, Jennifer S; Cabot, Ryan

    2017-12-01

    In vitro embryo production is an established method for both humans and animals, but is fraught with inferior development and health issues in offspring born after in vitro fertilization procedures. Analysis of epigenetic changes caused by exposure to in vitro conditions should shed light on potential sources of these phenotypes. Using immunocytochemistry, we investigated the localization and relative abundance of components associated with the SWI/SNF (Switch/Sucrose non-fermentable) chromatin-remodeling complex-including BAF155, BAF170, BAF180, BAF53A, BAF57, BAF60A, BAF45D, ARID1A, ARID1B, ARID2, SNF5, and BRD7-in oocytes and in in vitro-produced and in vivo-derived porcine embryos. Differences in the localization of BAF155, BAF170, BAF60A, and ARID1B among these sources indicate that improper timing of chromatin remodeling and cellular differentiation might occur in early preimplantation embryos produced and cultured in vitro. © 2017 The Authors. Molecular Reproduction and Development Published by Wiley Periodicals Inc.

  3. Metformin enhances the action of insulin on porcine granulosa-lutein cells in vitro.

    Science.gov (United States)

    Lee, Myeong Seop; Kim, Sang Hwan; Kim, Dae Seung; Min, Kwan Sik; Yoon, Jong Taek

    2012-12-01

    Metformin is an oral antidiabetic drug extensively used to treat the polycystic ovary syndrome in women. Metformin increases insulin-stimulated glucose uptake and has direct effects on ovarian steroidogenesis in humans. However, the molecular mechanisms of metformin' action on the ovary are not clear. To investigate the effects of this drug on the insulin-signaling pathway in porcine granulosa cells as an alternative model for human research, we examined the mRNA expressions of porcine insulin receptor (INSR), insulin-like growth factor-1 receptor (IGF-1R), insulin receptor substrate-1 (IRS-1), and the protein activity (activation and phosphorylation) of downstream targets including Raf, mitogen-activated protein kinase (MEK)1/2, extracellular signal regulated kinase (ERK), phosphoinositide-dependent 1 kinase (PDK1), mammalian target of rapamycin (TOR), p70, and nuclear factor-κB (NF-κB) in a primary culture system consisting of porcine granulosa-lutein cells (pGLs) incubated with 10(-5)M metformin and/or 100ng/ml insulin for 24h in a serum-free medium. We also investigated the luciferase activity of transcription factors activator protein-1 (AP-1) and NF-κB. Metformin with insulin significantly increased mRNA expressions of INSR, IGF-1R, and IRS-1, while metformin alone had no significant effect. And metformin with insulin had the significant effect on the protein activity (activation and phosphorylation) of downstream targets of INSR signaling pathway. Metformin with insulin significantly elicited an induction of luciferase activity in the transfection of AP-1 and NF-κBreporter, while metformin alone did not. In conclusion, we examined the activity of metformin and insulin on pGLS in vitro and metformin enhanced the action of insulin on the intracellular signaling pathways. These results suggest that metformin could change the function of ovarian granulosa cells. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Phenotypic and Functional Properties of Porcine Dedifferentiated Fat Cells during the Long-Term Culture In Vitro

    Directory of Open Access Journals (Sweden)

    Xuewu Peng

    2015-01-01

    Full Text Available It has been proved that terminally differentiated mature adipocytes possess abilities to dedifferentiate into fibroblast-like progeny cells with self-renewal and multiple differentiation, termed dedifferentiated fat (DFAT cells. However, the biological properties of DFAT cells during long-term culture in vitro have not been elucidated. Here, we obtained fibroblast-like morphology of porcine DFAT cells by ceiling culture. During the dedifferentiation process, round mature adipocytes with single large lipid droplets changed into spindle-shaped cells accompanied by the adipogenic markers PPARγ, aP2, LPL, and Adiponectin significant downregulation. Flow cytometric analysis showed that porcine DFAT cells displayed similar cell-surface antigen profile to mesenchymal stem cells (MSCs. Furthermore, different passages of porcine DFAT cells during long-term culture in vitro retained high levels of cell viabilities (>97%, efficient proliferative capacity including population doubling time ranged from 20 h to 22 h and population doubling reached 47.40±1.64 by 58 days of culture. In addition, porcine DFAT cells maintained the multiple differentiation capabilities into adipocytes, osteoblasts, and skeletal myocytes and displayed normal chromosomal karyotypes for prolonged passaging. Therefore, porcine DFAT cells may be a novel model of stem cells for studying the functions of gene in the different biological events.

  5. Involvement of bicarbonate-induced radical signaling in oxysterol formation and sterol depletion of capacitating mammalian sperm during in vitro fertilization.

    Science.gov (United States)

    Boerke, Arjan; Brouwers, Jos F; Olkkonen, Vesa M; van de Lest, Chris H A; Sostaric, Edita; Schoevers, Eric J; Helms, J Bernd; Gadella, Barend M

    2013-01-01

    This study demonstrates for the first time that porcine and mouse sperm incubated in capacitation media supplemented with bicarbonate produce oxysterols. The production is dependent on a reactive oxygen species (ROS) signaling pathway that is activated by bicarbonate and can be inhibited or blocked by addition of vitamin E or vitamin A or induced in absence of bicarbonate with pro-oxidants. The oxysterol formation was required to initiate albumin dependent depletion of 30% of the total free sterol and >50% of the formed oxysterols. Incubation of bicarbonate treated sperm with oxysterol-binding proteins (ORP-1 or ORP-2) caused a reduction of >70% of the formed oxysterols in the sperm pellet but no free sterol depletion. Interestingly, both ORP and albumin treatments led to similar signs of sperm capacitation: hyperactivated motility, tyrosin phosphorylation, and aggregation of flotillin in the apical ridge area of the sperm head. However, only albumin incubations led to high in vitro fertilization rates of the oocytes, whereas the ORP-1 and ORP-2 incubations did not. A pretreatment of sperm with vitamin E or A caused reduced in vitro fertilization rates with 47% and 100%, respectively. Artificial depletion of sterols mediated by methyl-beta cyclodextrin bypasses the bicarbonate ROS oxysterol signaling pathway but resulted only in low in vitro fertilization rates and oocyte degeneration. Thus, bicarbonate-induced ROS formation causes at the sperm surface oxysterol formation and a simultaneous activation of reverse sterol transport from the sperm surface, which appears to be required for efficient oocyte fertilization.

  6. Neonatal risks from in vitro fertilization and delayed motherhood.

    Science.gov (United States)

    Bellieni, Carlo Valerio

    2012-12-08

    Delayed childbearing (DC) is common in most Western countries. The average age of first-time mothers increased in United States from 21.4 years in 1970 to 25.0 years in 2006 and from 25.4 to 30.8 years in Australia in the same period. It is commonly believed that this has no ominous consequences. But several negative consequences of this behavior are described: stillbirth, prematurity, twins, birth anomalies. Age also decreases women's fertility, thus many couples undergo in vitro fertilization. And we highlight a paradox: medical reproduction techniques decreases their effectiveness with maternal age, but their availability can be an incentive to postpone parenthood. Of course the risks of delayed parenthood involve a minority of cases, but are parents entitled to accept any risk on the behalf of their baby A complete information would make people cautious before deciding to postpone childbearing, though this is often an obliged rather that a free choice: the consumerist society pressure and the difficulty to find an employment have their heavy weight in this choice. But if this choice is not really free, people's interest is to overcome these pressures and to claim for a real broad choice on when becoming parent, despite the pressures made by their cultural environment to postpone parenthood. Moreover, even reproductive techniques have some risks. Unfortunately, mass media often praise and endorse DC, disregarding the increase of premature babies born because of DC, a real alarm for public health. Pediatricians should discourage the culture that makes DC a normal event.

  7. Sperm penetration through the human zona pellucida as a predictor of in vitro fertilization.

    Science.gov (United States)

    Morales, P; Vantman, D; Madariaga, M

    1999-05-01

    The aim of this study was to determine the predictive value of sperm penetration into the perivitelline space of human cadaveric oocytes on in vitro fertilization outcome. Forty-two patients with tubal infertility undergoing ovarian stimulation with gonadotropin for in vitro fertilization and embryo transfer participated in the study. The number of spermatozoa bound to the human zona pellucida, the percentage of cadaveric oocytes with one or more spermatozoa in the perivitelline space, and the in vitro fertilization outcome were evaluated. Spermatozoa from 37 of 42 patients were able to penetrate the perivitelline space of cadaveric oocytes as well as to fertilize human oocytes in vitro. In three individuals, no penetration of the perivitelline space of cadaveric oocytes was observed and no in vitro fertilization was detected. Only two patients were able to fertilize the couple's oocytes without penetration of the cadaveric oocytes. Based on these results the specificity and the sensitivity of the assay to predict in vitro fertilization was 100% and 94.1%, respectively. Accordingly, these results suggest that sperm-zona penetration is a useful bioassay to predict male fertility potential in IVF outcome.

  8. Effect of punicalagin on proliferation of porcine ovarian granulosa cells in vitro

    Directory of Open Access Journals (Sweden)

    Dagmara Packová

    2016-12-01

    Full Text Available Punicalagin is a major component responsible for pomegranate's (Punica granatum antioxidant properties. Punicalagin is the predominant ellagitannin of Punica granatum and present in two isomeric forms: punicalagin α and β. Punicalagin is metabolised to ellagic acid (antioxidant and microorganisms present in colon can metabolize ellagic acid to urolithins. The aim of in vitro study was to examine the effect of punicalagin on mitochondrial activity and markers of proliferation in porcine ovarian granulosa cells. The cells were cultivated during 24h without (control group and with various doses (0.01, 0.1, 1, 10 and 100 μg*ml-1 of pomegranate compound – punicalagin. MTT assay and immunocytochemistry were used in this study. Stimulatory influence of punicalagin on the mitochondrial activity of ovarian granulosa cells at concentrations 1 μg*ml-1 was found. Punicalagin (at 1 μg*ml-1 had a significant (P < 0.05 impact on the presence of proliferative markers cyclin B1 (increase and PCNA - proliferating cell nuclear antigen (decrease in porcine ovarian granulosa cells. These results suggest dose-dependent effect of punicalagin on cell proliferation. Further verification of possible role of punicalagin in proliferation is therefore needed.

  9. 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

    The onset of ribosomal RNA (rRNA) synthesis occurs during the second half of the third cell cycle, that is, at the four-cell stage, in porcine embryos developed in vivo. In the present study the onset of rRNA synthesis was investigated in porcine embryos produced in vitro (IVP) or by somatic cell...... nuclear transfer (SCNT) using fluorescence in situ hybridization (FISH) with an rDNA probe and subsequent visualization of the nucleolar proteins by silver staining. In the 205 IVP embryos investigated, all two-cell embryos (n = 34) were categorized as transcriptionally inactive. At the late four...

  10. [In vitro fertilization program in therapy of the sterile marriage. International status and personal experiences].

    Science.gov (United States)

    Sudik, R; Fliess, F R; Wilken, H

    1984-01-01

    A review is given about in vitro fertilization (IVF/ET) and embryo transfer in the treatment of sterile couples. Programmes of in vitro fertilization of successful international groups are analysed and discussed, especially the improvements of the method. These international reports were the basis of the programme of in vitro fertilization at women's hospital of the Wilhelm Pieck University at Rostock. The organization of the programme depends on local conditions. Treatment is performed in some so-called IVF-series. The results of the first and second series with 52 patients are reported. The only pregnancy during the second series was an ectopic pregnancy.

  11. Keratoconus Progression Induced by In Vitro Fertilization Treatment.

    Science.gov (United States)

    Yuksel, Erdem; Yalinbas, Duygu; Aydin, Bahri; Bilgihan, Kamil

    2016-01-01

    To evaluate patients with keratoconus who manifested progression after in vitro fertilization (IVF) treatment. Patients with keratoconus who received IVF treatment were included in this study. None of the patients became pregnant as a result of the IVF treatment. Progression of keratoconus was determined by changes in corrected distance visual acuity and/or topographic changes and subjective assessments. Three patients with keratoconus received IVF treatment and keratoconus progression was detected in all 6 eyes of the patients. The mean age of the patients was 32.3 ± 3.6 years (range: 28 to 36 years) and the mean follow-up duration was 15.6 ± 3.2 months (range: 12 to 18 months). The mean and the maximum keratometry values increased and corrected distance visual acuity decreased after 2.3 IVF treatments. Drugs used in IVF treatment increase estrogen levels, which may affect corneal biomechanics and induce progression of keratoconus. Corneal cross-linking treatment could be offered to minimize the risk of keratoconus progression before IVF treatment. Copyright 2016, SLACK Incorporated.

  12. Isolation, Genome Phylogenetic Analysis and In vitro Rescue of a Newly Emerging Porcine Circovirus Type 2

    Directory of Open Access Journals (Sweden)

    Weijuan Zhu and Xiaofeng Ren*

    2012-05-01

    Full Text Available Porcine circovirus type 2 (PCV2 is the major causative agent of post-weaning multisystemic wasting syndrome (PMWS. Infection by PCV2 may cause heavy losses in pig industry. In this study, we report the isolation of a newly emerging PCV2 from northeastern China. The complete genome of the PCV2 isolate named PCV2-LJR contains 1766 nucleotides and was compared with reference sequences published in GenBank followed by topology analysis of the resulting phylogenetic tree. The data indicated that the prevalent PCV2 isolates in the northeastern China had close relationship, although various genotypes of PCV2 existed. In addition, by gene recombination and transfection techniques, the PCV2 infectious clone was achieved and was able to rescue virus in vitro determined by indirect immunofluorescence assay and PCR. The obtained biological materials may be used for biological characterization of PCV2.

  13. The fertilization ability and developmental competence of bovine oocytes grown in vitro.

    Science.gov (United States)

    Makita, Miho; Ueda, Mayuko; Miyano, Takashi

    2016-08-25

    In vitro growth culture systems for oocytes are being developed in several mammalian species. In these growth culture systems, in vitro grown oocytes usually have lower blastocyst formation than in vivo grown oocytes after in vitro fertilization. Furthermore, there have been a few reports that investigated the fertilization ability of in vitro grown oocytes in large animals. The purpose of this study was to investigate the fertilization process and developmental competence of bovine oocytes grown in vitro. Oocyte-granulosa cell complexes collected from bovine early antral follicles (0.4-0.7 mm in diameter) were cultured for growth with 17β-estradiol and androstenedione for 14 days and matured in vitro. These oocytes were then inseminated for 6 or 12 h, and further cultured for development up to 8 days in vitro. After growth culture, oocytes grew from 95 µm to around 120 µm and acquired maturation competence (79%). Although fertilization rates of in vitro grown oocytes were low after 6 h of insemination, 34% of in vitro grown oocytes fertilized normally after 12 h of insemination, having two polar bodies and two pronuclei with a sperm tail, and 22% of these oocytes developed into blastocysts after 8 days of culture. The fertilization and blastocyst formation rates were similar to those of in vivo grown oocytes. In addition, blastocyst cell numbers were also similar between in vitro and in vivo grown oocytes. In conclusion, in vitro grown bovine oocytes are similar to in vivo grown oocytes in fertilization ability and can develop into blastocysts.

  14. The fertilization ability and developmental competence of bovine oocytes grown in vitro

    Science.gov (United States)

    MAKITA, Miho; UEDA, Mayuko; MIYANO, Takashi

    2016-01-01

    In vitro growth culture systems for oocytes are being developed in several mammalian species. In these growth culture systems, in vitro grown oocytes usually have lower blastocyst formation than in vivo grown oocytes after in vitro fertilization. Furthermore, there have been a few reports that investigated the fertilization ability of in vitro grown oocytes in large animals. The purpose of this study was to investigate the fertilization process and developmental competence of bovine oocytes grown in vitro. Oocyte-granulosa cell complexes collected from bovine early antral follicles (0.4−0.7 mm in diameter) were cultured for growth with 17β-estradiol and androstenedione for 14 days and matured in vitro. These oocytes were then inseminated for 6 or 12 h, and further cultured for development up to 8 days in vitro. After growth culture, oocytes grew from 95 µm to around 120 µm and acquired maturation competence (79%). Although fertilization rates of in vitro grown oocytes were low after 6 h of insemination, 34% of in vitro grown oocytes fertilized normally after 12 h of insemination, having two polar bodies and two pronuclei with a sperm tail, and 22% of these oocytes developed into blastocysts after 8 days of culture. The fertilization and blastocyst formation rates were similar to those of in vivo grown oocytes. In addition, blastocyst cell numbers were also similar between in vitro and in vivo grown oocytes. In conclusion, in vitro grown bovine oocytes are similar to in vivo grown oocytes in fertilization ability and can develop into blastocysts. PMID:27151093

  15. Ejaculate and type of freezing extender affect rates of fertilization of horse oocytes in vitro.

    Science.gov (United States)

    Roasa, L M; Choi, Y H; Love, C C; Romo, S; Varner, D D; Hinrichs, K

    2007-09-01

    In vitro fertilization (IVF) was performed on in vitro-matured equine oocytes in three experiments. Frozen-thawed sperm were prepared using swim-up separation and heparin treatment. In Experiment 1, fertilization was achieved with sperm from only one frozen ejaculate of four obtained from the same stallion. Within this ejaculate, fertilization rates were higher with fresh media, as compared to media held for 6-8 days before use (39.6% versus 7.3%, respectively; Pfertilization rates (4% versus 39.6%; Pfertilization rates (range, 0-3%). In Experiment 3, fertilization rates of semen frozen in an extender containing 21.5% egg yolk were lower than fertilization rates of semen from the same ejaculate but frozen with a 3% egg-yolk extender (0% versus 15%, respectively; Pfertilization in this species. These factors may help to explain the great variability in fertilization rates reported with equine IVF, both among and within laboratories.

  16. Porcine sclera as a model of human sclera for in vitro transport experiments: histology, SEM, and comparative permeability

    Science.gov (United States)

    Ferrari, G.; Quarta, M.; Macaluso, C.; Govoni, P.; Dallatana, D.; Santi, P.

    2009-01-01

    Purpose To evaluate porcine sclera as a model of human sclera for in vitro studies of transscleral drug delivery of both low and high molecular weight compounds. Methods Human and porcine scleras were characterized for thickness and water content. The tissue surface was examined by scanning electron microscopy (SEM), and the histology was studied with hematoxylin-eosin staining. Comparative permeation experiments were performed using three model molecules, acetaminophen as the model compound for small molecules; a linear dextran with a molecular weight of 120 kDa as the model compound for high molecular weight drugs; and insulin, which was chosen as the model protein. Permeation parameters such as flux, lag time, and permeability coefficient were determined and compared. Results Human and porcine scleras have a similar histology and collagen bundle organization. The water content is approx 70% for both tissues while a statistically significant difference was found for the thickness, porcine sclera being approximately twofold thicker than human sclera. Differences in thickness produced differences in the permeability coefficient. In fact, human sclera was found to be two to threefold more permeable toward the three molecules studied than porcine sclera. Conclusions The results obtained in the present paper prove that porcine sclera can be considered a good model for human sclera for in vitro permeation experiments of both low and high molecular weight compounds. In fact, if the different tissue thickness is taken into account, comparable permeability was demonstrated. This suggests a possible use of this model in the evaluation of the transscleral permeation of new biotech compounds, which currently represent the most innovative and efficient therapeutic options for the treatment of ocular diseases. PMID:19190734

  17. Influence of cysteamine on in vitro maturation, in vitro and in vivo fertilization of equine oocytes.

    Science.gov (United States)

    Deleuze, S; Dubois, C S; Caillaud, M; Bruneau, B; Goudet, G; Duchamp, G

    2010-02-01

    Contents The effect of cysteamine on in vitro nuclear and cytoplasmic maturation of equine oocytes collected by transvaginal ultrasound guided follicular aspiration was assessed. Oocytes were matured in vitro with (cysteamine group) or without (control group) cysteamine. The nuclear stage after DNA Hoechst staining, penetration rates after two different in vitro fertilization (IVF) techniques (IVF media with ionophore and Hepes buffer with heparin) and the embryo yield following oocyte intra-oviductal transfer were used as a criterion for assessing nuclear and cytoplasmic maturation, respectively. Contrary to the data described in other domestic species, there was no effect of cysteamine on in vitro nuclear maturation, IVF or in vivo embryonic development under our conditions. Ovum pick up yields (52%) and maturation rates (control group: 47% and cysteamine group: 55%) were similar to those previously reported. From 57 oocytes transferred to the oviduct in each group, the number of embryos collected was 10 (17%) in the control group and five in the cysteamine group (9%). Those two percentages were not statistically different (p > 0.05). No effect of IVF technique was seen on the success rate (6%) in each group.

  18. In vivo and in vitro Anti-fertility and Anti-implantation Properties of ...

    African Journals Online (AJOL)

    The anti-fertility effect of the aqueous and ethanol extracts of the leaves and roots of Leonotis ocymifolia were studied both in vivo and in vitro. The anti-implantation and anti-fertility activities of the ethanol leaves extract were 37% and 20%, respectively. The body weight recorded for 19 days starting from day 1 of pregnancy ...

  19. Preterm and premature rupture of membranes in pregnancies after in vitro fertilization

    OpenAIRE

    Tabs Dunja; Vejnović Tihomir; Radunović Nebojša

    2005-01-01

    Women conceiving by assisted reproduction are at higher risk for preterm and premature rupture of membranes. The aim of our study was to estimate and compare incidence of preterm premature rupture of membranes in singleton pregnancies of women who conceived by intrauterine insemination and in vitro fertilization, from 1999 to 2003. We investigated 87 women from the intrauterine insemination, and 102 from the in vitro fertilization program. There were no statistically significant differences i...

  20. Poor ovarian response to stimulation for in vitro fertilization

    Directory of Open Access Journals (Sweden)

    Spremović-Rađenović Svetlana

    2015-01-01

    Full Text Available The term “poor respond (POR patients” is used for the group of women who respond badly to usual doses of gonadotropins in in vitro fertilization (IVF treatments; the consequence is low pregnancy rate. A consensus was reached on the minimal criteria needed to define POR. At least two of the following three features must be present: 1. advanced maternal age (40 years or more 2. previous POR (3 or less oocytes with a conventional stimulation protocol 3. abnormal ovarian reserve (AMH 0.5-1.1 ng/ml or AFC 5-7. The aim is to find better therapeutic options for these patients. Increased levels of day 3 follicle stimulating hormone (FSH and estradiol (E2, as well as decreased levels of anti-Mьllerian hormone (AMH and antral follicle count (AFC, can be used to assess ovarian reserve, as indirect predictive tests. A larger number of well designed, large scale, randomized, controlled trials are needed to assess the efficacy of different management strategies for poor responders: flare up gonadotropin releasing hormone (GnRH agonist protocols, modified long GnRH agonist mini-dose protocols, luteal initiation GnRH agonist stop protocol, pretreatment with estradiol - GnRH antagonist in luteal phase, natural cycle aspiration or natural cycle aspiration GnRH antagonist controlled, adjuvant therapy with growth hormone or dehydroepiandrosterone (DHEA. The results of up to now used protocols are unsatisfactory and stimulation of the ovulation in poor responders remains a challenge, especially when bearing in mind that in the majority of cases the patients will be menopausal in relatively short period of time.

  1. Expression analysis of pluripotency factors in the undifferentiated porcine inner cell mass and epiblast during in vitro culture.

    Science.gov (United States)

    Blomberg, Le Ann; Schreier, Lori L; Talbot, Neil C

    2008-03-01

    Limited understanding of the importance of known pluripotency factors in pig embryonic stem cells (ESC) impedes the establishment and validation of porcine ESC lines. This study evaluated the expression of known mouse ESC and human ESC (hESC) pluripotency markers in in vivo inner cell mass (ICM) and in vitro-cultured undifferentiated porcine epiblast cells isolated from 8-day porcine blastocysts, primary cultures of epiblast-derived neuroprogenitor cells, and endoderm cells. The expression profile of common pluripotency markers (POU domain 5 transcript factor 1, SRY-box containing gene 2, and Nanog homeobox), species-specific markers, ESC-associated factors, and differentiation markers was evaluated. The mRNA of uncultured ICMs, cultured epiblast cells, epiblast-derived neuroprogenitor cells, and endoderm cells was amplified prior to expression analysis of candidate genes by real-time RT-PCR. ESC factors whose expression correlated best with the undifferentiated epiblast state were identified by comparative mRNA expression analysis between porcine epiblast-derived somatic cell lines, fetal fibroblasts, and adult tissues. Across tissue types Nanog homeobox exhibited ubiquitous expression, whereas POU domain 5 transcript factor 1, teratocarcinoma-derived growth factor 1, and RNA exonuclease homolog 1 transcript expression was restricted primarily to undifferentiated epiblasts. Our results suggested that expression of pluripotency markers in undifferentiated pig epiblast cells more closely resembled that observed in hESC. Expression alterations of ESC-associated factors in epiblast cells were also observed during in vitro culture. Our data demonstrate the potential use of some pluripotency factors as markers of porcine epiblast stem cells and indicate that the in vitro environment may influence the cultured epiblast's developmental state. Copyright 2007 Wiley-Liss, Inc.

  2. [Relationships among human follicular fluid-induced acrosome reaction, sperm morphology and in vitro fertilization rates].

    Science.gov (United States)

    Li, Jian-ping; Zhong, Ying; Wu, Dong; Ai, Ling; Wang, Sheng; Tan, Chao; Zeng, Wei-qian; Liu, Jing; Ma, Guang-ping

    2006-07-01

    To assess the relationships among human follicular fluid-induced acrosome reaction, sperm morphology and in vitro fertilization rates. The relationships among human follicular fluid-induced acrosome reaction, sperm morphology and in vitro fertilization rates were investigated by Spearman rank correlation in 79 infertile couples. And the sperm morphology analysis was performed by crystal violet staining and based on strict criteria. A significant positive correlation was found between the percentage of human follicular fluid-induced acrosome reaction and that of normal sperm morphology (n = 49, r = 0.3763, P reaction and in vitro fertilization rates or between that of normal sperm morphology and in vitro fertilization rates (n = 21, r = 0.2666, P > 0.05 and n = 50, r = 0.0018, P > 0.05, respectively). There is a significant positive correlation between the percentage of human follicular fluid-induced acrosome reaction and that of normal sperm morphology, but no such correlation either between the percentage of human follicular fluid-induced acrosome reaction and in vitro fertilization rates or between that of normal sperm morphology and in vitro fertilization rates.

  3. EFFECT OF DEOXYNIVALENOL ON SOME HAEMATOLOGICAL, BIOCHEMICAL AND ANTIOXIDANT PARAMETERS OF PORCINE BLOOD IN VITRO

    Directory of Open Access Journals (Sweden)

    Katarína Zbyňovská

    2013-02-01

    Full Text Available The most important and the most common Fusarium mycotoxin is deoxynivalenol (DON. It occurs predominantly in grains such as wheat, barley, and maize and less often in oats, rice, rye, sorghum and triticale. It has adverse effects on humans, animals, and crops that result in illnesses and economic losses. The aim of the present study was to investigate the effect of DON on some haematological (red blood cells - RBC, white blood cells - WBC, platelets - PLT, haemoglobin - HGB, packed cell volume - PCV and lymphocyte - LYM, biochemical (cholesterol, triglycerides, total protein, urea, calcium and phosphorus and anti- and pro-oxidants parameters (superoxide dismutase - SOD, glutathione peroxidase - GPx and ROS – reactive oxygen species in porcine blood in vitro. Significantly decreased content of total protein in the group with dose of 1000 ng.l-1 DON was observed compared with the control group. In other groups (E1 with 10 ng.l-1 DON and E2 with 100 ng.l-1 slightly lower values were measures in comparison with the control group. PLT significantly decreased in the experimental group E3 when compared with E1, E2, and the control group. Concentration of GPx in porcine blood significantly (P < 0.05 decreased in E1 against the control group. Concentration of SOD significantly (P < 0.05 decreased in group E2 in comparison with E1 group. The highest value of ROS was in E2 group. Other parameters were not influenced by The most important and the most common Fusarium mycotoxin is deoxynivalenol (DON. It occurs predominantly in grains such as wheat, barley, and maize and less often in oats, rice, rye, sorghum and triticale. It has adverse effects on humans, animals, and crops that result in illnesses and economic losses. The aim of the present study was to investigate the effect of DON on some haematological (red blood cells - RBC, white blood cells - WBC, platelets - PLT, haemoglobin - HGB, packed cell volume - PCV and lymphocyte - LYM, biochemical

  4. Nuclear organization during in vitro differentiation of porcine mesenchymal stem cells (MSCs) into adipocytes.

    Science.gov (United States)

    Stachecka, Joanna; Walczak, Agnieszka; Kociucka, Beata; Ruszczycki, Błażej; Wilczyński, Grzegorz; Szczerbal, Izabela

    2018-02-01

    Differentiation of progenitor cells into adipocytes is accompanied by remarkable changes in cell morphology, cytoskeletal organization, and gene expression profile. Mature adipocytes are filled with a large lipid droplet and the nucleus tends to move to the cell periphery. It was hypothesized that the differentiation process is also associated with changes of nuclear organization. The aim of this study was to determine the number and distribution of selected components of nuclear architecture during porcine in vitro adipogenesis. The pig is an important animal model sharing many similarities to humans at the anatomical, physiological, and genetic levels and has been recognized as a good model for human obesity. Thus, understanding how cellular structures important for fundamental nuclear processes may be altered during adipocyte differentiation is of great importance. Mesenchymal stem cells (MSCs) were derived from bone marrow (BM-MSCs) and adipose tissue (AD-MSCs) and were cultured for 7 days in the adipogenic medium. A variable differentiation potential of these cell populations towards adipogenic lineage was observed, and for further study, a comparative characteristic of the nuclear organization in BM-MSCs and AD-MSCs was performed. Nuclear substructures were visualized by indirect immunofluorescence (nucleoli, nuclear speckles, PML bodies, lamins, and HP1α) or fluorescence in situ hybridization (telomeres) on fixed cells at 0, 3, 5, and 7 days of differentiation. Comprehensive characterization of these structures, in terms of their number, size, dynamics, and arrangement in three-dimensional space of the nucleus, was performed. It was found that during differentiation of porcine MSCs into adipocytes, changes of nuclear organization occurred and concerned: (1) the nuclear size and shape; (2) reduced lamin A/C expression; and (3) reorganization of chromocenters. Other elements of nuclear architecture such as nucleoli, SC-35 nuclear speckles, and telomeres

  5. Do Needleless Knots have Similar Strength as the Krackow Suture? An In Vitro Porcine Tendon Study.

    Science.gov (United States)

    Hong, Chih-Kai; Kuo, Ting-Hsuan; Yeh, Ming-Long; Jou, I-Ming; Lin, Cheng-Li; Su, Wei-Ren

    2017-02-01

    Numerous needleless techniques for tendon graft fixation that feature several advantages have been reported. However, there are few studies that have compared the holding strength between the needleless techniques (rolling hitch and modified rolling hitch) and traditional suture methods. To compare the tendon graft-holding strength of the rolling hitch and modified rolling hitch with the Krackow stitch in an in vitro porcine biomechanical model. Thirty fresh-frozen porcine flexor profundus tendons were randomly divided into three groups of 10 specimens. The experimental procedure was designed to assess elongation of the suture-tendon construct across the needleless tendon-grasping techniques and the Krackow stitch. All suture configurations were completed with a braided nonabsorbable suture. Each tendon was pretensioned to 100 N for three cycles, cyclically loaded from 50 to 200 N for 200 cycles, and then finally loaded to failure. Elongation, load to failure, and mode of failure were recorded for each specimen. Five of the 10 rolling hitch specimens failed during cyclic loading. With the numbers available, elongation after cyclic loading was not different among the successful rolling hitch specimens (19% [1.19 cm/6.17 cm] ± 6%), modified rolling hitch fixations (19% [1.11 cm/ 5.93 cm] ± 6%), and Krackow stitch fixations (26% [1.41 cm/5.43 cm] ± 6%); ultimate failure loads also were not different among the rolling hitch fixations (316 ± 35 N), modified rolling hitch fixations (342 ± 14 N), and Krackow stitches (327 ± 33 N) with the numbers available. With the numbers available, the rolling hitch, modified rolling hitch, and Krackow stitch techniques were not different in terms of elongation after cyclic loading and to failure in this in vitro biomechanical evaluation. Based on the biomechanical properties from this in vitro animal study, the modified rolling hitch may be an attractive alternative for tendon graft fixation in ligament-reconstruction surgery

  6. Constructing Human Skin Equivalents on Porcine Acellular Peritoneum Extracellular Matrix for In Vitro Irritation Testing.

    Science.gov (United States)

    Tsai, Pei-Chin; Zhang, Zheng; Florek, Charles; Michniak-Kohn, Bozena B

    2016-01-01

    The irritancy of topical products has to be investigated to ensure the safety and compliance. Although several reconstructed human epidermal models have been adopted by the Organization for Economic Cooperation and Development (OECD) to replace in vivo animal irritation testing, these models are based on a single cell type and lack dermal components, which may be insufficient to reflect all of the components of irritation. In our study, we investigated the use of acellular porcine peritoneum extracellular matrix as a substrate to construct full-thickness human skin equivalents (HSEs) for use as irritation screening tool. The acellular peritoneum matrix (APM) exhibited excellent skin cell attachment (>80%) and proliferation for human dermal fibroblasts (HDF) and immortalized human keratinocytes (HaCaT). APM-HSEs based on coculture of HDF and HaCaT were prepared. Increased HDF seeding density up to 5 × 10(4)/cm(2) resulted in APM-HSEs with a thicker and more organized epidermis. The epidermis of APM-HSEs expressed keratin 15, a keratinocyte proliferation marker, and involucrin, a differentiation marker, respectively. To assess the use of APM-HSEs for irritation testing, six proficiency chemicals, including three nonirritants (phosphate-buffered saline, polyethylene glycol 400, and isopropanol) and three irritants (1-bromohexane, heptanol, and sodium dodecyl sulfate) were applied. The APM-HSEs were able to discriminate nonirritants from irritants based on the viability. Levels of cytokines (interleukin [IL]-1α, IL-1ra, IL-6, IL-8, and granulocyte macrophage colony-stimulating factor [GM-CSF]) in these treatment groups further assisted the irritancy ranking. In conclusion, we have developed partially differentiated full-thickness APM-HSEs based on acellular porcine peritoneum matrix, and these APM-HSEs demonstrated utility as an in vitro irritation screening tool.

  7. The secretions of oviduct epithelial cells increase the equine in vitro fertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?

    Science.gov (United States)

    Mugnier, Sylvie; Kervella, Morgane; Douet, Cécile; Canepa, Sylvie; Pascal, Géraldine; Deleuze, Stefan; Duchamp, Guy; Monget, Philippe; Goudet, Ghylène

    2009-11-19

    Oviduct epithelial cells (OEC) co-culture promotes in vitro fertilization (IVF) in human, bovine and porcine species, but no data are available from equine species. Yet, despite numerous attempts, equine IVF rates remain low. Our first aim was to verify a beneficial effect of the OEC on equine IVF. In mammals, oviductal proteins have been shown to interact with gametes and play a role in fertilization. Thus, our second aim was to identify the proteins involved in fertilization in the horse. In the first experiment, we co-incubated fresh equine spermatozoa treated with calcium ionophore and in vitro matured equine oocytes with or without porcine OEC. We showed that the presence of OEC increases the IVF rates. In the subsequent experiments, we co-incubated equine gametes with OEC and we showed that the IVF rates were not significantly different between 1) gametes co-incubated with equine vs porcine OEC, 2) intact cumulus-oocyte complexes vs denuded oocytes, 3) OEC previously stimulated with human Chorionic Gonadotropin, Luteinizing Hormone and/or oestradiol vs non stimulated OEC, 4) in vivo vs in vitro matured oocytes. In order to identify the proteins responsible for the positive effect of OEC, we first searched for the presence of the genes encoding oviductin, osteopontin and atrial natriuretic peptide A (ANP A) in the equine genome. We showed that the genes coding for osteopontin and ANP A are present. But the one for oviductin either has become a pseudogene during evolution of horse genome or has been not well annotated in horse genome sequence. We then showed that osteopontin and ANP A proteins are present in the equine oviduct using a surface plasmon resonance biosensor, and we analyzed their expression during oestrus cycle by Western blot. Finally, we co-incubated equine gametes with or without purified osteopontin or synthesized ANP A. No significant effect of osteopontin or ANP A was observed, though osteopontin slightly increased the IVF rates. Our study

  8. The secretions of oviduct epithelial cells increase the equine in vitro fertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?

    Directory of Open Access Journals (Sweden)

    Canepa Sylvie

    2009-11-01

    Full Text Available Abstract Background Oviduct epithelial cells (OEC co-culture promotes in vitro fertilization (IVF in human, bovine and porcine species, but no data are available from equine species. Yet, despite numerous attempts, equine IVF rates remain low. Our first aim was to verify a beneficial effect of the OEC on equine IVF. In mammals, oviductal proteins have been shown to interact with gametes and play a role in fertilization. Thus, our second aim was to identify the proteins involved in fertilization in the horse. Methods & results In the first experiment, we co-incubated fresh equine spermatozoa treated with calcium ionophore and in vitro matured equine oocytes with or without porcine OEC. We showed that the presence of OEC increases the IVF rates. In the subsequent experiments, we co-incubated equine gametes with OEC and we showed that the IVF rates were not significantly different between 1 gametes co-incubated with equine vs porcine OEC, 2 intact cumulus-oocyte complexes vs denuded oocytes, 3 OEC previously stimulated with human Chorionic Gonadotropin, Luteinizing Hormone and/or oestradiol vs non stimulated OEC, 4 in vivo vs in vitro matured oocytes. In order to identify the proteins responsible for the positive effect of OEC, we first searched for the presence of the genes encoding oviductin, osteopontin and atrial natriuretic peptide A (ANP A in the equine genome. We showed that the genes coding for osteopontin and ANP A are present. But the one for oviductin either has become a pseudogene during evolution of horse genome or has been not well annotated in horse genome sequence. We then showed that osteopontin and ANP A proteins are present in the equine oviduct using a surface plasmon resonance biosensor, and we analyzed their expression during oestrus cycle by Western blot. Finally, we co-incubated equine gametes with or without purified osteopontin or synthesized ANP A. No significant effect of osteopontin or ANP A was observed, though

  9. Radiation-induced cleavage delay in mouse zygotes fertilized in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, T.; Takiuchi, H.; Okuyama, K.; Ohyama, H.

    1986-05-01

    Recently a complete culture system for the in vitro fertilized mouse eggs to the expanded blastocyst. With such an in vitro fertilization system, zygotes can be selected at a known stage, irradiated at a precise time after fertilization and their development observed periodically through the expanded blastocyst stage. In the present study, the authors examined the quantitative correlation between the first cleavage delay and the radiation dose using the same experimental system. The mouse zygotes fertilized in vitro (BC3F/sub 1/ eggs x ICR sperm) were irradiated with X-rays (10-100 R) at the pronuclear stage (4 hr after insemination). The zygotes were observed periodically at 30 min intervals from 16 to 24 hr after insemination. The results show that the first cleavage was delayed depending linearly on the X-ray dose.

  10. [Correlation between polyspermy and the outcome of in vitro fertilization-embryo transfer

    Science.gov (United States)

    Ye, Ying-Hui; Xing, Lan-Feng; Jin, Fan; Xu, Chen-Ming

    2002-06-01

    OBJECTIVE: To explore the influence of polyspermy on IVF outcomes in in vitro fertilization and embryo transfer(IVF-ET). METHODS: The data from 496 IVF-ET cycles and 5349 oocytes were analyzed retrospectively. A comparison of a number of fertility parameters with and without polyspermy was done. The fertility parameters were the number of oocytes retrieved, percentage of mature oocytes, fertilization rate, cleavage rate, occytes for ET, pregnancy rate. RESULTS: The percentage of mature occytes, fertilization rate, cleavage rate was 67.0 %,76.7 %and 95.6 %, respectively( Ppolyspermy(23.6 %),but with no statistical significance ( P>0.05). CONCLUSION: Polyspermic fertilization is correlated with improved oocyte receptibility to sperm and could be considered as an encouraging sign for the success of IVF.

  11. Culture of porcine luteal cells as a substrate for in vitro maturation of porcine cumulus oocyte complexes. Establishment and characterization

    Directory of Open Access Journals (Sweden)

    Teplitz MA

    2016-12-01

    Full Text Available The aim of this study was to establish and characterize the porcine luteal cells (PLC culture for the subsequent coculture with porcine COC. The final purpose is to promote the oocyte maturation. The PLC was established using corpora lutea obtained from slaughterhouse ovaries. Corpora lutea were dissected and luteal tissue submitted to a mechanical and enzymatic digestion with collagenase IV. The cell suspension was filtered and centrifuged and the cells obtained were diluted in 15 mL of DMEM-F12 supplemented media. Diluted cells were seeded in 3 culture flasks T25, staying in a controlled environment and changing the medium every 2 days. For the analysis and characterization, the cells were assessed by the Nile red staining to detect intracellular lipids, immunocytochemistry (ICC for 3β-hydroxy steroid dehidrogenase (3β-HSD and ELISA for P4 determination. We observed the presence of lipid intracellular droplets. Also, we observed an increase of P4 concentration at 48, 96 y 144 h of primary culture and almost all the cells were positive to the ICC evaluation for 3β-HSD, showing the steroidogenic capacity of the culture cells.

  12. Heat stress during in vitro fertilization decreases fertilization success by disrupting anti-polyspermy systems of the oocytes.

    Science.gov (United States)

    Sakatani, Miki; Yamanaka, Kenichi; Balboula, Ahmed Z; Takenouchi, Naoki; Takahashi, Masashi

    2015-01-01

    Low pregnancy rates during the summer are due, in part, to reduced fertilization. Given that elevated temperature is associated with this season, we investigated the effect of heat stress during fertilization using an in vitro model. Three experiments were performed to determine the mechanism by which exposure to elevated temperature disrupts fertilization. Oocytes were fertilized for 6 hr at 38.5°C or 41.0°C or 40.0°C with non-pre-incubated sperm, or for 6 hr at 38.5°C with sperm that had been pre-incubated at 38.5°C or 41.0°C for 4 hr. In each experiment, zygotes were cultured at 38.5°C in 5% CO(2) and 5% O(2). Rates of cleavage and blasocyst formation were reduced when fertilization occurs at elevated temperatures. The percent of sperm classified as alive, using fluorescein diacetate labeling, was decreased by pre-incubation and fertilization at 40.0°C. Although no difference was observed in sperm penetration rate, polyspermy tended to be increased by heat stress during fertilization. The zona pellucidae of zygotes formed following fertilization at 40.0°C for 6 hr were more sensitive to digestion with pronase. Furthermore, these zygotes exhibited higher hydrogen peroxide levels, measured by 2,7-dihydrodichlorofluorescein diacetate staining, and showed increased transcript abundance for HSPA1A, a gene involved in the heat-shock response, but decreased transcript abundance for UCHL1, a gene involved in preventing polyspermy. Results indicate that heat stress during fertilization is lethal to sperm, and causes oxidative stress, altered transcript abundance, and a defective block to polyspermy in the zygote. Thus, an increase in polyspermy is likely one cause of the reduced competency of zygotes fertilized under elevated temperatures to develop to the blastocyst stage. © 2014 Wiley Periodicals, Inc.

  13. EFFECT OF dbcAMP ON PROLIFERATION AND APOPTOSIS OF PORCINE GRANULOSA CELLS in vitro

    Directory of Open Access Journals (Sweden)

    Richard Alexa

    2013-02-01

    Full Text Available Cyclic nucleotide cAMP and its target protein kinase A (PKA dependent intracellular mechanisms can play an important role in regulation of ovarian cell function and in mediating gonadotropin action on these cells. The aim of the present study was to examine the effect of cAMP analogue, dibutyryl cyclic adenosine monophosphate (dbcAMP (0; 0.1; 1 and 10 µg/ml or FSH (0; 0,01; 1 IU/ml on proliferation and apoptosis of porcine granulosa cells in vitro. Indices of cell apoptosis (expression of apoptotic peptide bax and proliferation (expression of proliferation-associated peptide PCNA within ovarian granulosa cells were analysed by immunocytochemistry. It was observed that accumulation of PCNA was increased by dbcAMP and FSH at all doses added. The occurrence of bax was also stimulated by dbcAMP after exposition (at 0,1 and 1 µg/ml, but not at dose 10 µg/ml and by FSH (at all doses added. The stimulatory effect of both dbcAMP and FSH on both ovarian cell apoptosis and proliferation suggest, that these substances may promote ovarian follicular cell turnover. The similarity of dbcAMP and FSH effect may indicate that FSH can affect ovarian functions via cAMP-dependent intracellular mechanisms. The present data may provide new tools to regulate human and animal reproductive processes via cAMP-dependent mechanisms.

  14. DOSE-DEPENDENT EFFECT OF MOLYBDENUM ON PORCINE BLOOD CELLS: IN VITRO ASSESSMENT

    Directory of Open Access Journals (Sweden)

    Marcela Capcarová

    2013-10-01

    Full Text Available The aim of this study was to examine the effect of molybdenum (Mo on selected haematological parameters of porcine blood in vitro. The samples of blood were treated with an ammonium molybdate (NH46.Mo7O24.4H2O for 4 hours at 37oC in the concentrations 10, 100, and 1000 µg.ml-1 (E1, E2, and E3 group. Blood without Mo addition served as the control (C. Selected haematological parameters (WBC – total white blood cell count, LYM – lymphocyte count, GRA – granulocyte count, RBC – red blood cell count, HGB – haemoglobin, HCT – haematocrit and PLT – platelet count were measured using haematological analyser Abacus junior VET. Significant decrease in WBC, LYM, and GRA in the group with the highest dose of Mo (E3 against other groups (C, E1, and E2 was observed. Other results were not influenced by Mo exposure. Our results suggest that Mo can cause the changes and imbalance in immune system.

  15. Experimental factors affecting in vitro absorption of six model compounds across porcine skin.

    Science.gov (United States)

    Karadzovska, Daniela; Brooks, James D; Riviere, Jim E

    2012-10-01

    This comparative study evaluated the effect of several experimental variables on the absorption of six model [(14)C]-labeled compounds (caffeine, cortisone, diclofenac sodium, mannitol, salicylic acid, and testosterone) through porcine skin. Using static and flow-through diffusion cells, finite or infinite, saturated or unsaturated doses were applied in one of three vehicles: propylene glycol, water, and ethanol following a full factorial experimental design. The flux of each compound into the receptor phase, with or without bovine serum albumin (BSA), was monitored over 24 h. Levels of radioactivity were also determined in the stratum corneum by tape stripping and in the remaining skin. Apparent permeability coefficients (Kp) and dose absorbed were calculated and compared. The overall results emphasize the importance of experimental design and confirm previous findings that identified dose volume, saturation level and vehicle as the main sources of variation in the in vitro assessment of dermal absorption, whilst diffusion cell model and the presence/absence of BSA in the receptor phase had minimal effect. Although the acquired data do not directly reveal new mechanistic information on dermal absorption, the unique and complete study design has provided a suitable data source for the development of dermal absorption prediction models. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. The secretory products of Trichomonas vaginalis decrease fertilizing capacity of mice sperm in vitro

    Science.gov (United States)

    Roh, Jaesook; Lim, Young-Su; Seo, Min-Young; Choi, Yuri; Ryu, Jae-Sook

    2015-01-01

    Trichomonas vaginalis infection is one of the most prevalent sexually transmitted infections in humans and is now recognized as an important cause of infertility in men. There is little information about the effect of extracellular polymeric substances (EPS) from T. vaginalis on sperm, but previous reports do not provide a conclusive description of the functional integrity of the sperm. To investigate the impact of EPS on the fertilizing capacity of sperm, we assessed sperm motility, acrosomal status, hypo-osmotic swelling, and in vitro fertilization rate after incubating the sperm with EPS in vitro using mice. The incubation of sperm with EPS significantly decreased sperm motility, viability, and functional integrity in a concentration and time-dependent manner. These effects on sperm quality also resulted in a decreased fertilization rate in vitro. This is the first report that demonstrates the direct negative impact of the EPS of T. vaginalis on the fertilization rate of sperm in vitro. However, further study should be performed using human sperm to determine if EPS has similar negative impact on human sperm fertilizing capacity in vitro. PMID:25578937

  17. In vitro fertilizing potential of urethral and epididymal spermatozoa collected from domestic cats (Felis catus).

    Science.gov (United States)

    Prochowska, S; Niżański, W

    2017-03-28

    The aim of this study was to provide a comparative analysis of in vitro fertilizing potential of frozen-thawed urethral and epididymal feline spermatozoa. Both types of semen were collected from 7 cats and cryopreserved in liquid nitrogen. To perform in vitro fertilization, both urethral and epididymal samples from the same individual were thawed and spermatozoa were co-incubated with in vitro matured cat oocytes. Obtained embryos were cultured in vitro for 7 days in a commercial medium. Cleavage rate, morula rate and blastocyst rate were calculated. Experiment was run in 10 replicates. The examined parameters showed no significant differences between urethral and epididymal spermatozoa (p>0.05). Cleavage rate and embryo's development were highly variable between replicates, even for the different sperm samples collected from one individual. There was no significant correlation between fertilizing capacity of two types of spermatozoa collected from the same male. In this study we confirmed that cryopreserved urethral spermatozoa have equally good fertilizing potential as epididymal ones, and both can be successfully used for in vitro fertilization in cats with the use of commercial medium.

  18. Urinary paraben concentrations and in vitro fertilization outcomes among women from a fertility clinic

    Science.gov (United States)

    Mínguez-Alarcón, Lidia; Chiu, Yu-Han; Messerlian, Carmen; Williams, Paige L.; Sabatini, Mary E.; Toth, Thomas L.; Ford, Jennifer B.; Calafat, Antonia M.; Hauser, Russ

    2015-01-01

    Objective To explore the relationship between urinary paraben concentrations and IVF outcomes among women attending an academic fertility center. Design Prospective cohort study. Setting Fertility clinic in a hospital setting. Patient(s) A total of 245 women contributing 356 IVF cycles. Intervention(s) None. Quantification of urinary concentrations of parabens by isotope-dilution tandem mass spectrometry, and assessment of clinical endpoints of IVF treatments abstracted from electronic medical records at the academic fertility center. Main Outcome Measure(s) Total and mature oocyte counts, proportion of high quality embryos, fertilization rates, and rates of implantation, clinical pregnancy and live births. Results The geometric mean of the urinary concentrations of methyl (MP), propyl (PP) and butyl paraben (BP) in our study population were 133, 24 and 1.5 µg/L, respectively. In models adjusted for age, body mass index, race/ethnicity, smoking status and primary infertility diagnosis, urinary MP, PP and BP concentrations were not associated with IVF outcomes, specifically total and mature oocyte counts, proportion of high embryo quality and fertilization rates. Moreover, no significant associations were found between urinary paraben concentrations and rates of implantation, clinical pregnancy and live births. Conclusion(s) Urinary paraben concentrations were not associated with IVF outcomes among women undergoing infertility treatments. PMID:26654974

  19. Urinary paraben concentrations and in vitro fertilization outcomes among women from a fertility clinic.

    Science.gov (United States)

    Mínguez-Alarcón, Lidia; Chiu, Yu-Han; Messerlian, Carmen; Williams, Paige L; Sabatini, Mary E; Toth, Thomas L; Ford, Jennifer B; Calafat, Antonia M; Hauser, Russ

    2016-03-01

    To explore the relationship between urinary paraben concentrations and IVF outcomes among women attending an academic fertility center. Prospective cohort study. Fertility clinic in a hospital setting. A total of 245 women contributing 356 IVF cycles. None. Quantification of urinary concentrations of parabens by isotope-dilution tandem mass spectrometry, and assessment of clinical endpoints of IVF treatments abstracted from electronic medical records at the academic fertility center. Total and mature oocyte counts, proportion of high-quality embryos, fertilization rates, and rates of implantation, clinical pregnancy, and live births. The geometric means of the urinary concentrations of methylparaben, propylparaben, and butylparaben in our study population were 133, 24, and 1.5 μg/L, respectively. In models adjusted for age, body mass index, race/ethnicity, smoking status, and primary infertility diagnosis, urinary methylparaben, propylparaben, and butylparaben concentrations were not associated with IVF outcomes, specifically total and mature oocyte counts, proportion of high embryo quality, and fertilization rates. Moreover, no significant associations were found between urinary paraben concentrations and rates of implantation, clinical pregnancy, and live births. Urinary paraben concentrations were not associated with IVF outcomes among women undergoing infertility treatments. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. Assessment of the antiviral properties of recombinant porcine SP-D against various influenza A viruses in vitro.

    Directory of Open Access Journals (Sweden)

    Marine L B Hillaire

    Full Text Available The emergence of influenza viruses resistant to existing classes of antiviral drugs raises concern and there is a need for novel antiviral agents that could be used therapeutically or prophylacticaly. Surfactant protein D (SP-D belongs to the family of C-type lectins which are important effector molecules of the innate immune system with activity against bacteria and viruses, including influenza viruses. In the present study we evaluated the potential of recombinant porcine SP-D as an antiviral agent against influenza A viruses (IAVs in vitro. To determine the range of antiviral activity, thirty IAVs of the subtypes H1N1, H3N2 and H5N1 that originated from birds, pigs and humans were selected and tested for their sensitivity to recombinant SP-D. Using these viruses it was shown by hemagglutination inhibition assay, that recombinant porcine SP-D was more potent than recombinant human SP-D and that especially higher order oligomeric forms of SP-D had the strongest antiviral activity. Porcine SP-D was active against a broad range of IAV strains and neutralized a variety of H1N1 and H3N2 IAVs, including 2009 pandemic H1N1 viruses. Using tissue sections of ferret and human trachea, we demonstrated that recombinant porcine SP-D prevented attachment of human seasonal H1N1 and H3N2 virus to receptors on epithelial cells of the upper respiratory tract. It was concluded that recombinant porcine SP-D holds promise as a novel antiviral agent against influenza and further development and evaluation in vivo seems warranted.

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

    African Journals Online (AJOL)

    Marqueta J, Pareja A, et al. Impact of the Spanish Fertility. Society guidelines on the number of embryos to transfer. Reprod Biomed Online 2010;21:667‑75. 21. Ryan GL, Sparks AE, Sipe CS, Syrop CH, Dokras A, Van. Voorhis BJ. A mandatory single blastocyst transfer policy with educational campaign in a United States ...

  2. High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification

    DEFF Research Database (Denmark)

    Du, Y; Pribenszky, C S; Molnár, M

    2008-01-01

    The purpose of the present study was to improve cryotolerance using high hydrostatic pressure (HHP) pretreatment of porcine in vitro matured (IVM) oocytes, to facilitate their further developmental competence after parthenogenetic activation. A total of 1668 porcine IVM oocytes were used in our...... present study. The pressure tolerance and optimal duration of recovery after HHP treatment were determined. Oocytes were treated with either 20 or 40 MPa (200 and 400 times greater than atmospheric pressure) for 60 min, with an interval of 10, 70, and 130 min between pressure treatment and subsequent...... vitrification under each pressure parameter. Oocytes from all vitrification groups had much lower developmental competence than fresh oocytes (Ppressure, with either 70...

  3. Ethical and Legal Aspects of the Fertilization in Vitro: Case Artavia Murillo Vs. Costa Rica

    Directory of Open Access Journals (Sweden)

    Helena Cristina Aguiar De Paula Vilela

    2016-10-01

    Full Text Available This paper analyzes the Case Fertilization in vitro vs. Costa Rica, judged by the Inter- American Court of Human Rights, from the perspective of bioethics and law. For this, the theories about the beginnings of life were discussed, as the theoretical debate of the Kantian imperative and the Beck's risk society, in order to provide the discussion on human dignity and risks of research in biotechnology and genetics. For the IACHR,to prohibit fertilization in vitro in Costa Rica was discriminatory and violated theduty of not arbitrary interference in private life and the right of family’s formation.

  4. [In vitro fertilization and embryo transfer. Personal experiences in the introduction of laboratory methodology].

    Science.gov (United States)

    Fliess, F R; Sudik, R

    1984-01-01

    Our laboratory technique includes recovery and handling with gametes outside the body, in vitro fertilization, culture of embryos, and finally embryo-transfer. Time-table, culture media and technique of the cultivation as well as laboratory devices are described and than compared with informations about methods from other teams. The necessity of transport of the follicle fluids from operating theatre to laboratory is given caused by local conditions. All dates about number of oocytes (73), quantities of preovulatory oocytes (47) and cleavage rates (55%) are descended from 30 laparoscopic oocyte recoveries of the 2. in vitro fertilization program of the university woman hospital in Rostock, January and February 1984.

  5. Polyspermy in in vitro fertilization of human oocytes: frequency and possible causes.

    Science.gov (United States)

    van der Ven, H H; Al-Hasani, S; Diedrich, K; Hamerich, U; Lehmann, F; Krebs, D

    1985-01-01

    In an IVF program a total of 585 oocytes (180 patients) were examined for the presence of pronuclei 16 to 20 hours after the addition of spermatozoa. The overall fertilization rate was 71%, and in 58 (10%) of the fertilized oocytes, three or more pronuclei, indicating a failure of the block to polyspermy, could be observed. The frequency of polyspermy was related to the maturity of the oocyte, determined according to morphologic criteria. Immature oocytes showed a higher percentage of polyspermic fertilization (32%) compared to that of mature oocytes (6%). Preincubation of oocytes (for 0.5-1.5, 2-4, and 5-8 hours) prior to the addition of spermatozoa increased the fertilization rate (to 67%, 70%, and 83%, respectively). The polyspermy rate, however, was not significantly different between the various preincubation intervals (13%, 14%, and 19%, respectively). The polyspermy rate was affected by the number of spermatozoa used for in vitro fertilization. Insemination with 0.5-0.8, 1.0, or 1.5-2.0 X 10(6) spermatozoa/oocyte resulted in a polyspermy rate of 6%, 20%, and 32%, respectively. The appearance of polyspermic fertilization was not related to the age of the patient (which ranged from 20 to 45 years) nor to the method of ovarian stimulation (clomiphene, hMG, or clomiphene/hMG). Because of the high incidence of polyspermy under in vitro conditions it seems to be important to routinely examine the oocytes in the pronuclear stage. Reduction of the number of spermatozoa used for in vitro fertilization and the exact timing of insemination according to the maturity of the oocyte might reduce the occurrence of polyspermic fertilization.

  6. In vitro measurements of optical properties of porcine brain using a novel compact device

    CSIR Research Space (South Africa)

    Yavari, N

    2005-09-01

    Full Text Available describes measurements of the optical properties of porcine brain tissue using novel instrumentation for simultaneous absorption and scattering characterisation of small turbid samples. Integrating sphere measurements are widely used as a reference method...

  7. In vitro porcine blastocyst development in three-dimentional alginate hydrogels

    Science.gov (United States)

    Appropriate embryonic and fetal development significantly impact pregnancy success and, therefore, the efficiency of swine production. The pre-implantation period of porcine pregnancy is characterized by several developmental hallmarks, which are initiated by the dramatic morphological change that o...

  8. Effect of tendon tensioning: an in vitro study in porcine extensor tendons.

    Science.gov (United States)

    Figueroa, David; Calvo, Rafael; Vaisman, Alex; Meleán, Patricio; Figueroa, Francisco

    2010-06-01

    Graft tensioning is a controversial issue in anterior cruciate ligament reconstruction (ACLR) that has not achieved consensus between peers. The purpose of this study is to determine if after tensioning tendon length and resistance to maximal load changes. We performed an in vitro study with 50 porcine extensors tendons. The first group (P=25) was tensioned with 80 N (19.97 lb) for 10 min, using an ACL graft preparation board. The second group (C=25) was used as control and was not tensioned. The average initial (groups P and C) and post tensioning tendon length (group C) were measured; the average initial and post tensioning tendon diameter were measured as well. All samples were fixated in a tube-clamp system connected to a tension sensor. The samples were stressed with continuous and progressive tension until ultimate failure at maximum load (UFML) occurs. The initial mean length was: P before tensioning=13.4 mm+/-1.4 mm (range 10.5-16.5); P after tensioning=13.8 mm+/-1.4 mm (range 11.5-16.5); C=13 mm+/-1.35 mm (p=0.005). The mean diameter was: P=5.6 mm (4.5-6); C=5.5 mm (range 4.5-6) (p>0.05). The UFML was: P=189.7 N (114-336); C=229.9 N (143-365) (p=0.029). Tendon tensioning with 80 N for 10 min produced 3% average elongation. These could be beneficial in ACLR since tendon tensioning decreases elongation of the graft after fixation. Regardless, tendon tensioning is not innocuous since it diminishes their resistance when continuously stressed until complete failure occurs.

  9. Comparison of solvents for removing pesticides from skin using an in vitro porcine model.

    Science.gov (United States)

    Campbell, J L; Smith, M A; Eiteman, M A; Williams, P L; Boeniger, M F

    2000-01-01

    This study compared four solvents (1-propanol, polyethylene glycol [avg. MW 400], 10% Ivory Liquid and water, and D-TAM) for their ability to remove selected pesticides from an in vitro porcine skin model using a solvent-moistened wipe. Wipes were performed 90 min after pesticide was applied to the skin. The four pesticides selected (glyphosate, alachlor, methyl parathion, and trifluralin) were chosen because of their differences in water solubility. This study also determined whether pretreatment of skin with a solvent prior to pesticide application would either increase or decrease recovery of the pesticide. Recovery efficiencies for all solvents and pesticides were affected by the amount of contaminant on the skin. Although pesticide recoveries from all four solvents were similar (range: 45-57%), on average 1-propanol had significantly higher recoveries, followed by soap and water. There was no significant difference between polyethylene glycol, and D-TAM. When skin was pretreated with any of the four solvents before pesticide application, the recoveries of the more water soluble compounds, glyphosate and alachlor, decreased. When pretreatment with solvent preceded application of trifluralin, the pesticide with the lowest water solubility, recoveries increased. 1-Propanol or soap and water were more effective in removing pesticides from skin than polyethylene glycol or D-TAM, but the amount of pesticide recovered from skin was affected by the chemical characteristics of the pesticide (such as water solubility) and the amount of pesticide originally on the skin. This study provides information useful to the interpretation of skin wipe sample results collected in field studies.

  10. Effects of transferrin on aromatase activity in porcine granulosa cells in vitro.

    Directory of Open Access Journals (Sweden)

    Małgorzata Duda

    2009-01-01

    Full Text Available Proliferating cells have an absolute requirement for iron, which is delivered by transferrin with subsequent intracellular transport via the transferrin receptor. Recent studies have reported that transferrin plays a crucial role in the local regulation of ovarian function, apart from its iron-binding characteristic. Therefore, the present study was undertaken to explore the possible role of transferrin in porcine granulosa cells function by examining its influence on aromatase activity, the most important indicator of follicular cell differentiation. In the first series of studies, pig granulosa cells isolated from small, immature follicles were cultured in the presence of transferrin alone (10 microg/ml or 100 microg/ml or with the addition of FSH (100ng/ml. The second series of studies was undertaken to determine transferrin-stimulated granulosa cells ability to aromatize exogenous testosterone (1x10(-7M. One hour after the establishment of cultures an aromatase inhibitor CGS16949A was added to test its influence on estradiol production. After 48 hours, cultures were terminated and cells were processed for immunocytochemical staining of aromatase. Media were frozen for further estradiol level analysis. Positive immunostaining for aromatase was found in all granulosa cell cultures. The intensity of immunostaining was always stronger in cultures supplemented with FSH whereas the addition of transferrin had no effect. Granulosa cells in vitro synthesized the highest amount of estradiol after the addition of FSH and exogenous testosterone as measured radioimmunologically. Concomitant treatment with FSH and transferrin caused an inhibition of FSH-stimulated aromatase activity. The production of estradiol also declined in the presence of FSH, testosterone and transferrin. This study demonstrates that transferrin had a dose-dependent inhibitory effect on FSH-stimulated aromatase activity, which was confirmed by radioimmunoassay. Our results indicate

  11. Chemical composition and in vitro digestibility of whole-crop maize fertilized with synthetic fertilizer or digestate and harvested at two maturity stages in Boreal growing conditions

    National Research Council Canada - National Science Library

    Mahmoud F. Seleiman; Shaimaa Selim; Seija Jaakkola; Pirjo S.A. Mäkelä

    2017-01-01

    .... Effects of digestate (sludge from biogas of domestic origin) application in comparison with synthetic fertilizer and two maturity stages on chemical composition and in vitro digestibility of whole-crop maize were investigated...

  12. In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer.

    Science.gov (United States)

    Hinrichs, K; Love, C C; Brinsko, S P; Choi, Y H; Varner, D D

    2002-07-01

    Three experiments were conducted to evaluate the effect of oocyte and sperm treatments on rates of in vitro fertilization (IVF) in the horse and to determine the capacity of in vitro-matured horse oocytes to be fertilized in vivo. There was no effect of duration of oocyte maturation (24 vs. 42 h) or calcium ionophore concentration during sperm capacitation (3 microM vs. 7.14 microM) on in vitro fertilization rates. Oocytes matured in 100% follicular fluid had significantly higher fertilization (13% to 24%) than did oocytes matured in maturation medium or in 20% follicular fluid (0% to 12%; P fertilization rate among 3 sperm treatments utilizing 7.14 microM calcium ionophore (12% to 21%). Of in vitro-matured oocytes recovered 40-44 h after transfer to the oviducts of inseminated mares, 77% showed normal fertilization (2 pronuclei to normal cleavage). Cleavage to 2 or more cells was seen in 22% of oocytes matured in follicular fluid and 63% of oocytes matured in maturation medium; this difference was significant (P horse oocytes are capable of being fertilized at high rates in the appropriate environment and that in vitro maturation of oocytes in follicular fluid increases fertilization rate in vitro but reduces embryo development after fertilization in vivo. Further work is needed to determine the optimum environment for sperm capacitation and IVF in the horse.

  13. Release of EPA and DHA from salmon oil - a comparison of in vitro digestion with human and porcine gastrointestinal enzymes.

    Science.gov (United States)

    Aarak, K E; Kirkhus, B; Holm, H; Vogt, G; Jacobsen, M; Vegarud, G E

    2013-10-01

    In the present study, we hypothesised whether in vitro digestion of salmon oil would release different amounts of PUFA depending on the origin of the lipolytic enzymes used. For this purpose, in vitro digestion of salmon oil (SO) was performed using human duodenal juice (HDJ) or a commercial enzyme preparation consisting of porcine pancreatin and bile (PB). The lipolytic effect was determined by measuring the release of fatty acids (FA) using solid-phase extraction and GC-flame ionisation detection, withdrawing samples every 20 min during digestion. The amount of FA released indicated that a plateau was reached after 80 min with approximately similar amounts of FA detected using both HDJ and PB (379 (sd 18) and 352 (sd 23) mg/g SO, respectively). However, the release of 18 : 2, EPA (20 : 5) and DHA (22 : 6) was significantly different during in vitro digestion. At 80 min, HDJ and PB released 43 and 33% of 18 : 2, 14 and 9% of EPA and 11 and 9% of DHA, respectively. Both enzyme preparations released approximately the same amounts of the other FA analysed. The effect of the addition of bile salts (BS) was significantly different in the two enzyme systems, where porcine pancreatin highly responded to the increase in BS concentration, in contrast to HDJ.

  14. New Reproductive Assemblages: Understanding, Managing and ‘Using’ Human In Vitro Fertilization (IVF)

    NARCIS (Netherlands)

    Just, E.M.

    2009-01-01

    This dissertation is a contribution to the ongoing discussion about the body and in vitro fertilization (IVF), also known as assisted reproduction or technologically enhanced reproduction. With help of empirical research on Dutch and Polish IVF-couples, Edyta Just puts into question the meaning of

  15. Effect of antral follicle count in beef heifers on in vitro fertilization/production

    Science.gov (United States)

    Our objective has been to compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers. This is the 4th year of the study with years 1 to 3 reported individually. For this report, we add data for the 4th year and present a combined an...

  16. Prediction of pregnancy success rate through in vitro fertilization based on maternal age

    Directory of Open Access Journals (Sweden)

    Soegiharto Soebijanto

    2009-12-01

    Full Text Available Aim To evaluate the correlation between the success of pregnancy through in vitro fertilization and maternal age. Methods Assessment of pregnancy was performed in eight in vitro fertilization centers in Indonesia: Harapan Kita Pediatric and Obstetric Hospital from 1997 to 2001, and seven in vitro fertilization centers in Indonesia. Follicular induction was performed through the long protocol, short protocol and natural cycle. Insemination was performed through ICSI (intra cytoplasmic sperm injection on petri dish. Spermatozoa were obtained through masturbation, testicular biopsy and epididimical biopsy. A successful pregnancy was indicated chemically, with the presence of fetal heart beat and the birth of a baby (take home baby. Results There was a 34% pregnancy rate for the age group below 30 years, 33.75% for those between 31 and 35 years olds, and 26% for the age group 36 to 40 years old, and 8% for the age group above 40 years. Conclusion The higher the maternal age, the lower pregnancy rate. In other words, the higher the maternal age, the higher the rate of miscarriage. (Med J Indones 2009; 18: 244-8Keywords: pregnancy, in vitro fertilization

  17. In vitro fertilization pregnancy in a patient with proven chronic endometritis

    NARCIS (Netherlands)

    Fatemi, Human Mousavi; Popovic-Todorovic, Biljana; Ameryckx, Linda; Bourgain, Claire; Fauser, Bart; Devroey, Paul

    Objective: To report an in vitro fertilization (IVF) pregnancy in a patient with histologically confirmed chronic endometritis before the IVF treatment without prior antibiotherapy. Design: Case report. Setting: Academic reproductive medicine unit. Patient(s): A30-year-oldwoman with primary

  18. Neurological Condition of Infants Born After In Vitro Fertilization With Preimplantation Genetic Screening

    NARCIS (Netherlands)

    Middelburg, Karin J.; Heineman, Maas J.; Haadsma, Maaike L.; Bos, Arend F.; Kok, Joke H.; Hadders-Algra, Mijna

    Aim of this study was to evaluate the effect of preimplantation genetic screening (PGS) on neurodevelopmental outcome in children. We conducted a prospective follow-up Study of children born to women randomly assigned to in vitro fertilization with or without PGS. Primary outcome was adverse

  19. Neurological Condition of Infants Born After In Vitro Fertilization With Preimplantation Genetic Screening

    NARCIS (Netherlands)

    Middelburg, Karin J.; Jan Heineman, Maas; Haadsma, Maaike L.; Bos, Arend F.; Kok, Joke H.; Hadders-Algra, Mijna

    2010-01-01

    Aim of this study was to evaluate the effect of preimplantation genetic screening (PGS) on neurodevelopmental outcome in children. We conducted a prospective follow-up Study of children born to women randomly assigned to in vitro fertilization with or without PGS. Primary outcome was adverse

  20. A detailed cost analysis of in vitro fertilization and intracytoplasmic sperm injection treatment.

    NARCIS (Netherlands)

    Bouwmans, C.A.; Lintsen, B.M.; Eijkemans, M.J.; Habbema, J.D.; Braat, D.D.M.; Hakkaart, L.

    2008-01-01

    OBJECTIVE: To provide detailed information about costs of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment stages and to estimate the cost per IVF and ICSI treatment cycle and ongoing pregnancy. DESIGN: Descriptive micro-costing study. SETTING: Four Dutch IVF

  1. Ovarian Stimulation for In Vitro Fertilization and Long-term Risk of Breast Cancer

    NARCIS (Netherlands)

    Belt-Dusebout, A.W. van den; Spaan, M.; Lambalk, C.B.; Kortman, M.; Laven, J.S.; Santbrink, E.J. van; Westerlaken, L.A. van der; Cohlen, B.J.; Braat, D.D.M.; Smeenk, J.M.; Land, J.A.; Goddijn, M.; Golde, R.J. van; Rumste, M.M. van; Schats, R.; Jozwiak, K.; Hauptmann, M.; Rookus, M.A.; Burger, C.W.; Leeuwen, F.E. van

    2016-01-01

    IMPORTANCE: Previous studies of breast cancer risk after in vitro fertilization (IVF) treatment were inconclusive due to limited follow-up. OBJECTIVE: To assess long-term risk of breast cancer after ovarian stimulation for IVF. DESIGN, SETTING, AND PARTICIPANTS: Historical cohort (OMEGA study) with

  2. Does Spinal Analgesia have Advantage over General Anesthesia for Achieving Success in In-Vitro Fertilization?

    Science.gov (United States)

    Aghaamoo, Shahrzad; Azmoodeh, Azra; Yousefshahi, Fardin; Berjis, Katayon; Ahmady, Farahnazsadat; Qods, Kamran; Mirmohammadkhani, Majid

    2014-03-01

    Because of high psychological burden and considerable costs of in-vitro fertilization, it is greatly important to identify all factors that may influence its results. In this study, general anesthesia and spinal analgesia used for oocyte retrieval were compared in terms of success in treating infertility among couples who had undergone in-vitro fertilization at an infertility center in Tehran, Iran. This cohort study that was based on analysis of patient records at Mirza Kochak Khan Hospital, Tehran University of Medical Sciences, in 2008-2009. In this study, the status of chemical pregnancy among those who experienced general anesthesia or spinal anesthesia for in-vitro fertilization for the first time were compared, and the possible effects of clinical and laboratory factors using logistic regression models were considered. Considering the number of transferred embryos, underlying cause of infertility and fetus grade, it was found that practicing spinal anesthesia is significantly related to increased chance of chemical pregnancy (adjusted Odds Ratio=2.07; 95% CI: 1.02,4.20; p=0.043). According to analysis of recorded data in an infertility treatment center in Iran, it is recommended to use spinal anesthesia instead of general anesthesia for oocyte retrieval to achieve successful in-vitro fertilization outcome. This can be studied and investigated further via a proper multicentric study in the country.

  3. Economic value of in vitro fertilization in Ukraine, Belarus, and Kazakhstan

    NARCIS (Netherlands)

    E.A. Mandrik (Olena); S. Knies (Saskia); J.L. Severens (Hans)

    2015-01-01

    markdownabstract__Abstract__ Background: An economic value calculation was performed to estimate the lifetime net present value of in vitro fertilization (IVF) in Ukraine, Belarus, and Kazakhstan. Methods: Net lifetime tax revenues were used to represent governmental benefits accruing from a

  4. In vitro permeation and disposition of niacinamide in silicone and porcine skin of skin barrier-mimetic formulations.

    Science.gov (United States)

    Haque, Tasnuva; Lane, Majella E; Sil, Bruno C; Crowther, Jonathan M; Moore, David J

    2017-03-30

    Niacinamide (NIA) is an amide form of vitamin B3 which is used in cosmetic formulations to improve various skin conditions and it has also been shown to increase stratum corneum thickness following repeated application. In this study, three doses (5, 20 and 50μL per cm 2 ) of two NIA containing oil-in-water skin barrier-mimetic formulations were evaluated in silicone membrane and porcine ear skin and compared with a commercial control formulation. Permeation studies were conducted over 24h in Franz cells and at the end of the experiment membranes were washed and niacinamide was extracted. For the three doses, retention or deposition of NIA was generally higher in porcine skin compared with silicone membrane, consistent with the hydrophilic nature of the active. Despite the control containing a higher amount of active, comparable amounts of NIA were deposited in skin for all formulations for all doses; total skin absorption values (permeation and retention) of NIA were also comparable across all formulations. For infinite (50μL) and finite (5μL) doses the absolute permeation of NIA from the control formulation was significantly higher in porcine skin compared with both test formulations. This likely reflects differences in formulation components and/or presence of skin penetration enhancers in the formulations. Higher permeation for the 50 and 20μL dose was also evident in porcine skin compared with silicone membrane but the opposite is the case for the finite dose. The findings point to the critical importance of dose and occlusion when evaluating topical formulations in vitro and also the likelihood of exaggerated effects of excipients on permeation at infinite and pseudo-finite dose applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Paternal urinary concentrations of organophosphate flame retardant metabolites, fertility measures, and pregnancy outcomes among couples undergoing in vitro fertilization.

    Science.gov (United States)

    Carignan, Courtney C; Mínguez-Alarcón, Lidia; Williams, Paige L; Meeker, John D; Stapleton, Heather M; Butt, Craig M; Toth, Thomas L; Ford, Jennifer B; Hauser, Russ

    2018-02-01

    Use of organophosphate flame retardants (PFRs) has increased over the past decade following the phase out of some brominated flame retardants, leading to increased human exposure. We recently reported that increasing maternal PFR exposure is associated with poorer pregnancy outcomes among women from a fertility clinic. Because a small epidemiologic study previously reported an inverse association between male PFR exposures and sperm motility, we sought to examine associations of paternal urinary concentrations of PFR metabolites and their partner's pregnancy outcomes. This analysis included 201 couples enrolled in the Environment and Reproductive Health (EARTH) prospective cohort study (2005-2015) who provided one or two urine samples per IVF cycle. In both the male and female partner, we measured five urinary PFR metabolites [bis(1,3-dichloro-2-propyl) phosphate (BDCIPP), diphenyl phosphate (DPHP), isopropylphenyl phenyl phosphate (ip-PPP), tert-butylphenyl phenyl phosphate (tb-PPP) and bis(1-chloro-2-propyl) phosphate (BCIPP)] using negative electrospray ionization liquid chromatography tandem mass spectrometry (LC-MS/MS). The sum of the molar concentrations of the urinary PFR metabolites was calculated. We used multivariable generalized linear mixed models to evaluate the association of urinary concentrations of paternal PFR metabolites with IVF outcomes, accounting for multiple in vitro fertilization (IVF) cycles per couple. Models were adjusted for year of IVF treatment cycle, primary infertility diagnosis, and maternal urinary PFR metabolites as well as paternal and maternal age, body mass index, and race/ethnicity. Detection rates were high for paternal urinary concentrations of BDCIPP (84%), DPHP (87%) and ip-PPP (76%) but low for tb-PPP (12%) and zero for BCIPP (0%). We observed a significant 12% decline in the proportion of fertilized oocytes from the first to second quartile of male urinary ΣPFR and a 47% decline in the number of best quality embryos

  6. Effect of oviduct epithelial cells on the fertilization and development of sheep oocytes in vitro

    DEFF Research Database (Denmark)

    Holm, Peter; Irvine, Brendon J.; Armstrong, David T.

    1994-01-01

    cells and then transferred to the oviducts of a recipient ewe, 30 h post oestrus, for a 6.5 day period of in vivo culture. Similar rates of fertilization ( 54-58%) and blastocyst development from cleaving zygotes ( 48-69%) were achieved in both experi- ments in vitro with no evident benefit of including...... oviductal cells. In fact, fewer (P= 0 .02) blastocysts developed from cleaved embryos when co-cultured for the 96 h period (Group 4). Whilst the blastocyst development rates obtained in vitro ( 45-58%) were similar to, or higher than (P=0.03 ), those obtained in vivo ( 43%), more than 50% of in vitro cul...

  7. The secretory products of Trichomonas vaginalis decrease fertilizing capacity of mice sperm in vitro

    Directory of Open Access Journals (Sweden)

    Jaesook Roh

    2015-04-01

    Full Text Available Trichomonas vaginalis infection is one of the most prevalent sexually transmitted infections in humans and is now recognized as an important cause of infertility in men. There is little information about the effect of extracellular polymeric substances (EPS from T. vaginalis on sperm, but previous reports do not provide a conclusive description of the functional integrity of the sperm. To investigate the impact of EPS on the fertilizing capacity of sperm, we assessed sperm motility, acrosomal status, hypo-osmotic swelling, and in vitrofertilization rate after incubating the sperm with EPS in vitrousing mice. The incubation of sperm with EPS significantly decreased sperm motility, viability, and functional integrity in a concentration and time-dependent manner. These effects on sperm quality also resulted in a decreased fertilization rate in vitro. This is the first report that demonstrates the direct negative impact of the EPS of T. vaginalis on the fertilization rate of sperm in vitro. However, further study should be performed using human sperm to determine if EPS has similar negative impact on human sperm fertilizing capacity in vitro.

  8. [Is an act of human love the in vitro fertilization? A proposal ethical analysis].

    Science.gov (United States)

    García Sánchez, Emilio

    2014-01-01

    Since 1978, when the first test tube baby, Louis Brown, was born, thousands of children have been born every year through in vitro fertilization. Many families keep attending fertility clinics in order to receive some treatment for their infertility problems and have a child. Children born in this way are worthy human beings. Their parents love them and devote themselves to their children admirably, showing real parental love. However, does this loving kindness justify, from an ethical point of view, any way of desiring and having a son or daughter? Is it really an act of human love to long for a child and satisfy this desire using artificial methods? Is it equally human and worthy to wish them choosing in vitro fertilization than to wish them through an intimate and loving relationship, in which the child emerges as a result of interpersonal donation? I answer these questions by analyzing the ethics proposal formulated by Rhonheimer and Carrasco de Paula. In short, only the intimate and loving sexual union between a man and a woman -as long as it is unconditional love- may be the dignity cause of the existence of a human being. And such union and unconditional requirement are absent in vitro fertilization.

  9. Developmental potential of vitrified holstein cattle embryos fertilized in vitro with sex-sorted sperm.

    Science.gov (United States)

    Xu, J; Guo, Z; Su, L; Nedambale, T L; Zhang, J; Schenk, J; Moreno, J F; Dinnyés, A; Ji, W; Tian, X C; Yang, X; Du, F

    2006-07-01

    In vitro fertilization (IVF) is a feasible way to utilize sex-sorted sperm to produce offspring of a predetermined sex in the livestock industry. The objective of the present study was to examine the effects of various factors on bovine IVF and to systematically improve the efficiency of IVF production using sex-sorted sperm. Both bulls and sorting contributed to the variability among differential development rates of embryos fertilized by sexed sperm. Increased sorting pressures (275.8 to 344.75 kPa) did not have a significant effect on the in vitro fertility of the sorted sperm; neither did an extended period of 9 to 14 h from semen collection to sorting. As few as 600 sorted sperm were used to fertilize an oocyte, resulting in blastocyst development of 33.2%. Postwarming of vitrified sexed IVF embryos resulted in high morphological survival (96.3%) and hatching (84.4%) rates, similar to those fertilized by nonsexed sperm (93.1 and 80.6%, respectively). A 40.9% pregnancy rate was established following the transfer of 3,627 vitrified, sexed embryos into synchronized recipients. This was not different from the rates with nonsexed IVF (41.9%, n = 481), or in vivo-produced (53.1%, n = 192) embryos. Of 458 calves born, 442 (96.5%) were female and 99.6% appeared normal. These technologies (sperm sexing-IVF-vitrification-embryo transfer) provide farmers, as well as the livestock industry, with a valuable option for herd expansion and heifer replacement programs. In summary, calves were produced using embryos fertilized by sex-sorted sperm in vitro and cryopreserved by rapid cooling vitrification.

  10. Pig Spermatozoa Defect in Acrosome Formation Caused Poor Motion Parameters and Fertilization Failure through Artificial Insemination and In vitro Fertilization.

    Science.gov (United States)

    Lee, Won Young; Lee, Ran; Kim, Hee Chan; Lee, Kyung Hoon; Cui, Xiang Shun; Kim, Nam Hyung; Kim, Sang Hyun; Lee, Il Joo; Uhm, Sang Jun; Yoon, Min Jung; Song, Hyuk

    2014-10-01

    The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not

  11. Pig Spermatozoa Defect in Acrosome Formation Caused Poor Motion Parameters and Fertilization Failure through Artificial Insemination and In vitro Fertilization

    Science.gov (United States)

    Lee, Won Young; Lee, Ran; Kim, Hee Chan; Lee, Kyung Hoon; Cui, Xiang Shun; Kim, Nam Hyung; Kim, Sang Hyun; Lee, Il Joo; Uhm, Sang Jun; Yoon, Min Jung; Song, Hyuk

    2014-01-01

    The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not

  12. Full in vitro fertilization laboratory mechanization: toward robotic assisted reproduction?

    Science.gov (United States)

    Meseguer, Marcos; Kruhne, Ulrich; Laursen, Steen

    2012-06-01

    To describe the current efforts made to standardize different steps of assisted reproductive technology processes by the introduction of new technologies for the nonsubjective sperm selection process, oocyte denudation by mechanical removal of cumulus cells, oocyte positioning, sperm motility screening, fertilization, embryo culture, media replacement by microfluidics, and monitoring of embryo development by time-lapse photography, embryo secretions, and/or O(2) consumption. These technologies could be integrated in a unique and fully automated device. Pubmed database and research and development data from authors. University-affiliated private center. None. None. None. Several technologies would be useful for: 1) selection of sperm based on viability; 2) manipulation and removal of the cumulus cells' narrow channel regions combined with microfluidics; 3) advances in oocyte positioning precision through the use of joystick-controlled micromanipulators; 4) microfluidics allowing the gradual change of a culture medium, which might result in better embryo development as well as reduce the amount of embryo manipulation; 5) time-lapse, proteomic, and metabolic scoring of the developing embryo, allowing multiple and optimized selection of the embryos. The technologies described in this review have not yet reported reliable clinical proofs. We already have available some of the technologies described, but we envisage an integrated device, i.e., an IVF lab-on-a-chip, by which oocyte and sperm would be processed to achieve a perfect embryo ready to be delivered into the uterus. With such a device, sample preparation, chemical or biologic reactions, and data collection would be integrated. Copyright © 2012. Published by Elsevier Inc.

  13. In vitro engineering of a palatal mucosa equivalent with acellular porcine dermal matrix.

    Science.gov (United States)

    Xiong, Xuepeng; Zhao, Yifang; Zhang, Wei; Xie, Weiguo; He, Sangang

    2008-08-01

    The objective of this study was to develop a palatal mucosa equivalent composed of multilayered oral keratinocytes grown on the acellular porcine dermal matrix. Acellular porcine dermal matrix was prepared through a series of procedures and assessed by histological, immunohistochemical, and scanning electron microscopy examination. The palatal mucosa equivalent was fabricated by seeding oral keratinocytes, which cultured from human palate mucosa, onto the acellular dermal matrix. After 4 days submerged in medium, this composite was raised to the air-liquid interface for another 7 or 14 days of cultivation. The results demonstrated the processed porcine dermal matrix was totally cell-free. The resultant palatal mucosa equivalent showed a multilayered oral epithelium that had been formed, and the number of cell layers was correlated with the culture period at the air-liquid interface. Oral keratinocytes infiltrated into the empty hair follicles of the acellular porcine dermal matrix and formed an anchor-like structure, which exhibited resemblance to the rete ridges of the native palate mucosa. Immunohistochemical staining for CK10/13, CK19, Ki-67 nuclear antigen, and Heparan sulphate indicated the cultured palatal mucosa equivalent shared the same characteristics with that of the native palate mucosa. In conclusion, our fabricated palatal mucosa equivalent exhibited the characteristics of the native counterpart, and this equivalent might be useful for recovery of the wounds in the palate secondary to palatoplasty.

  14. Nicotine permeability across the buccal TR146 cell culture model and porcine buccal mucosa in vitro

    DEFF Research Database (Denmark)

    Nielsen, Hanne Mørck; Rassing, Margrethe Rømer

    2002-01-01

    comparable for bi-directional and uni-directional transport in the presence of a transmembrane pH gradient. Nicotine concentrations between 10(-5) and 10(-2) M were applied to the apical side of the TR146 cell culture model or the mucosal side of porcine buccal mucosa. Buffers with pH values of 5.5, 7...

  15. Artificial oocyte activation in intracytoplasmic sperm injection cycles using testicular sperm in human in vitro fertilization.

    Science.gov (United States)

    Kang, Hee Jung; Lee, Sun-Hee; Park, Yong-Seog; Lim, Chun Kyu; Ko, Duck Sung; Yang, Kwang Moon; Park, Dong-Wook

    2015-06-01

    Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n=480; AOA, n=29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n=12) and non-motile sperm-injected (n=17) groups (total n=29 cycles). The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or delivery rate (5.9% vs. 16.7%) between the two groups. We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure.

  16. In vitro fertilization rate of mouse eggs with sperm after X-irradiation at various spermatogenetic stages

    Energy Technology Data Exchange (ETDEWEB)

    Matsuda, Y.; Tobari, I.; Yamada, T. (National Inst. of Radiological Sciences, Chiba (Japan))

    1985-02-01

    The frequency of in vitro fertilization of mouse eggs using sperm obtained weekly (for 1-9 weeks) from epididymis after testicular X-irradiation (200 rad) was observed. The sperm concentrations for insemination were 100, 200 and 300/mm/sup 3/. The number of fertilized eggs seemed to remain constant almost at control level (90-98%) until the 4th week after X-irradiation. Thereafter, the number declined to reach a minimum level (about 30-50%) in the 6th week; they then recovered completely in the 8th week. This tendency was found at all sperm concentrations used. This result indicates that the most sensitive stage for the fertilization capacity of sperm to X-rays was the early spermatocytes or the late spermatogonia. The high radiosensitivity of the fertilization capacity of sperm irradiated during the early-spermatocyte to late-spermatogonial stage, corresponded well with the rate of induction of sperm abnormalities and spermatogenetic cell killing. The in vitro fertilization rate was not restored by an increase in sperm concentration at insemination; it is, therefore, likely that the in vitro fertilization frequency depends on the proportion of sperm with abnormal morphology and abnormal fertilization ability, and sperm number seems to have hardly any influence on the fertilization rate, as far as the in vitro fertilization experiment is concerned.

  17. In vitro fertilization and polyspermy in the pig: factors affecting fertilization rates and cytoskeletal reorganization of the oocyte.

    Science.gov (United States)

    Suzuki, Hiroyuki; Saito, Yosuke; Kagawa, Noriko; Yang, Xiangzhong

    2003-07-01

    Polyspermy is a common phenomenon in the pig. Extensive information has become available from in vitro studies on not only the quality of oocytes but also the quality of spermatozoa. However, little information is available on the relative penetration rates of fresh and frozen spermatozoa from the same ejaculate from boars of different breeds. The present results, based on a total of 15 boars of three different breeds, revealed that the inter-breed variation in fertilization and polyspermic rates is larger than intra-breed variation. It was also shown that the incidence of polyspermy as well as penetration rate was greatly decreased by freezing and thawing, even if a higher number of sperm was coincubated with cumulus-free oocytes for a longer period compared to fresh sperm of the same ejaculate. This study focuses on the cytoskeletal organization of the oocyte with respect to the status of cumulus investment, and monospermic and polyspermic fertilization. The status of cumulus cells correlated with the density of transzonal cumulus-cell processes and with the maturation rate of oocytes and, to some degrees, the incidence of polyspermy. Polyspermic zygotes formed multiple microtubule domains in association with individual male pronuclei (PN), but in a high degree of polyspermy (more than trispermy), the pronuclear apposition did not proceed. The effect of multiple PN of paternal and maternal origin on the cytoskeletal reorganization is also discussed. Copyright 2003 Wiley-Liss, Inc.

  18. The role of oxidative stress in female infertility and in vitro fertilization.

    Science.gov (United States)

    Wojsiat, Joanna; Korczyński, Jerzy; Borowiecka, Marta; Żbikowska, Halina Małgorzata

    2017-05-09

    Infertility problem involves many couples of reproductive age. It has been estimated that in Poland 0.7-1.0 million pairs require treatment, while for more than half of them assisted reproduction is the only recommended and effective method. Infertility affects 13 to 15% of the world's population. A major concern is the age-related decline in female fertility even more that often a decision about pregnancy is taken at later age. Recent studies show that increased production of reactive oxygen species is an important factor in etiopathogenesis of pregnancy and affects female reproduction. It was found that oxidative stress may damage the oocytes and may impair their fertilization capacity. Oxidative stress may also lead to embryo fragmentation and formation of numerous developmental abnormalities, and is regarded to be one of the important reasons of spontaneous and recurrent miscarriage. Moreover, overproduction of reactive oxygen species has a significant impact on the success of in vitro fertilization (IVF).

  19. Spontaneous fertility and in vitro fertilization outcome: new evidence of human papillomavirus sperm infection.

    Science.gov (United States)

    Garolla, Andrea; Engl, Bruno; Pizzol, Damiano; Ghezzi, Marco; Bertoldo, Alessandro; Bottacin, Alberto; Noventa, Marco; Foresta, Carlo

    2016-01-01

    To evaluate the reproductive outcome of infertile couples undergoing assisted reproduction techniques (ART) with or without human papillomavirus (HPV) semen infection. Cross-sectional clinical study. Units of andrology, reproductive medicine, and gynecology. A total of 226 infertile couples. Male partners were evaluated by means of fluorescence in situ hybridization (FISH) for HPV on semen. After a diagnostic period, female partners underwent intrauterine insemination (IUI) or intracytoplasmic sperm injection (ICSI). Seminal parameters and FISH analysis for HPV in sperm head. Spontaneous or assisted pregnancies, live births, and miscarriages were recorded. Statistical analysis included unpaired Student t test and chi-square test. Fifty-four male partners (23.9%) had HPV semen infection confined to sperm, confined to exfoliated cells, or in both cells. During the diagnostic period, noninfected couples showed spontaneous pregnancies. IUI and ICSI treatments were performed in, respectively, 60 and 98 noninfected and in 21 and 33 infected couples, with 38.4% and 14.2% cumulative pregnancy rates, respectively. The follow-up of pregnancies showed a higher miscarriage rate in infected couples (62.5% vs. 16.7%). Ongoing pregnancies of the latter group were characterized by HPV infection confined to exfoliated cells. A reduction in natural and assisted cumulative pregnancy rate and an increase in miscarriage rate are related to the presence of HPV at sperm level. Although the exact mechanism by which sperm infection is able to impair fertility remains unclear, this aspect is worthy of further investigations. If confirmed, these results could change the clinical and diagnostic approach to infertile couples. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. In vitro investigation of the follicular penetration of porcine ear skin using a nanoparticle-emulsion containing the antiseptic polihexanide In vitro investigation of the follicular penetration of porcine ear skin

    Science.gov (United States)

    Ulmer, M.; Patzelt, A.; Vergou, T.; Lademann, J.; Richter, H.; Kramer, A.; Müller, G.; Sterry, W.; Lange-Asschenfeldt, B.

    2012-05-01

    Earlier investigations regarding the distribution of the bacterial flora on the human skin demonstrate that the hair follicle acts as a bacterial reservoir, providing a quick source for secondary recontamination. These findings highlight the importance of the hair follicle as a target for modern antiseptics. In the present study, we have assessed the follicular penetration of a curcumin-labeled particle-associated antiseptic into porcine skin by laser scanning microscopy. Therefore, the follicular penetration depth of the curcumin-labeled particle-associated antiseptic was compared to the follicular penetration depth of curcumin-labeled particles without antiseptic. The investigation was performed in vitro using porcine skin biopsies. By superposition of the images acquired in the transmission and the fluorescent modus, it was possible to visualize the distribution of the fluorescent dye inside the hair follicles. Quantitative and qualitative results showed that both dispersions penetrated efficiently into the hair follicles. The average penetration depth of the particles with attached antiseptic polihexanide was significantly higher than that of particles without the attached antiseptic. Also, whilst very little sample preparation was needed, laser scanning microscopy was found to be an efficient tool to visualize the skin relief and in particular the hair follicle shaft and localize fluorescent markers within the skin tissue and hair follicles.

  1. OPEN PULLED STRAW (OPS) VITRIFICATION OF PORCINE BLASTOCYSTS: SIMPLE PROCEDURE YIELDING EXCELLENT IN VITRO SURVIVAL, BUT SO FAR NO PIGLETS FOLLOWING TRANSFER

    DEFF Research Database (Denmark)

    Holm, P; Vajta, G; Macháty, Z

    1999-01-01

    In 2 trials, 346 porcine in vivo blastocysts were vitrified in heat-softened and pulled French straws by the Open Pulled Straw (OPS) vitrification method. After thawing, embryos were either cultured in vitro (n=89) for 24 to 48 hours or transferred surgically (n=257) to 11 synchronised recipients...

  2. Live Births from Domestic Dog (Canis familiaris Embryos Produced by In Vitro Fertilization.

    Directory of Open Access Journals (Sweden)

    Jennifer B Nagashima

    Full Text Available Development of assisted reproductive technologies (ART in the dog has resisted progress for decades, due to their unique reproductive physiology. This lack of progress is remarkable given the critical role ART could play in conserving endangered canid species or eradicating heritable disease through gene-editing technologies-an approach that would also advance the dog as a biomedical model. Over 350 heritable disorders/traits in dogs are homologous with human conditions, almost twice the number of any other species. Here we report the first live births from in vitro fertilized embryos in the dog. Adding to the practical significance, these embryos had also been cryopreserved. Changes in handling of both gametes enabled this progress. The medium previously used to capacitate sperm excluded magnesium because it delayed spontaneous acrosome exocytosis. We found that magnesium significantly enhanced sperm hyperactivation and ability to undergo physiologically-induced acrosome exocytosis, two functions essential to fertilize an egg. Unlike other mammals, dogs ovulate a primary oocyte, which reaches metaphase II on Days 4-5 after the luteinizing hormone (LH surge. We found that only on Day 6 are oocytes consistently able to be fertilized. In vitro fertilization of Day 6 oocytes with sperm capacitated in medium supplemented with magnesium resulted in high rates of embryo development (78.8%, n = 146. Intra-oviductal transfer of nineteen cryopreserved, in vitro fertilization (IVF-derived embryos resulted in seven live, healthy puppies. Development of IVF enables modern genetic approaches to be applied more efficiently in dogs, and for gamete rescue to conserve endangered canid species.

  3. Risk of bowel obstruction during in vitro fertilization treatment of patients with deep infiltrating endometriosis.

    Science.gov (United States)

    Seyer-Hansen, Mikkel; Egekvist, Anne; Forman, Axel; Riiskjaer, Mads

    2018-01-01

    Women with endometriosis often experience pain and infertility. Medical treatment interferes with the possibility of attaining pregnancy. For infertile women with endometriosis, surgery is a possible treatment, but with advanced disease there is an increased risk of serious complications. With only limited pain, women will often be referred for in vitro fertilization treatment instead. The disease is estrogen-dependent and during in vitro fertilization treatment the women could theoretically experience worsening of their symptoms. The study is a retrospective cohort study of 76 women with bowel endometriosis who were treated conservatively and underwent in vitro fertilization treatment. Nine (11.8%) of the women experienced severe worsening of their bowel-related symptoms, including two patients presenting with colon ileus. One additional woman had no previous diagnosis of endometriosis before she presented with subocclusion of the bowel during in vitro fertilization. In all cases the in vitro fertilization treatment was stopped. Our study revealed that bowel endometriosis increases the risk of complications during in vitro fertilization treatment. This is in contrast to several publications. However, our study population is different due to the fact that none of these women had previous operations for bowel endometriosis. In all, 88% of the women completed fertility treatment without need for surgery. © 2017 Nordic Federation of Societies of Obstetrics and Gynecology.

  4. In vitro fertilization and artificial activation of eggs of the direct-developing anuran Eleutherodactylus coqui

    Directory of Open Access Journals (Sweden)

    Toro Esteban

    2004-08-01

    Full Text Available Abstract Although much is known about the reproductive biology of pond-breeding frogs, there is comparatively little information about terrestrial-breeding anurans, a highly successful and diverse group. This study investigates the activation and in vitro fertilization of eggs of the Puerto Rican coqui frog obtained by hormonally induced ovulation. We report that spontaneous activation occurs in 34% of eggs, probably in response to mechanical stress during oviposition. Artificial activation, as evidenced by the slow block to polyspermy and the onset of zygote division, was elicited both by mechanical stimulation and calcium ionophore exposure in 64% and 83% of the cases, respectively. Finally, one in vitro fertilization protocol showed a 27% success rate, despite the fact that about one third of all unfertilized eggs obtained by hormone injection auto-activate. We expect these findings to aid in the conservation effort of Eleutherodactylus frogs, the largest vertebrate genus.

  5. Filariasis infection is a probable cause of implantation failure in in vitro fertilization cycles.

    Science.gov (United States)

    Bazi, Tony; Finan, Ramzi; Zourob, Dani; Sabbagh, Amira S; Nasnas, Roy; Zreik, Tony G

    2006-06-01

    To describe a parasitic infection that probably affected the implantation of good-quality embryos in an in vitro fertilization (IVF) cycle. Case report. Tertiary care center in a university hospital. A 36-year-old Caucasian female with primary unexplained infertility. The patient underwent two cycles of IVF with good-quality embryos transferred; however, no pregnancy ensued despite adequate luteal support. In vitro fertilization cycles, CBC, blood smear, evaluation for eosinophilia including serological evaluation for parasitic infections. Pregnancy. Following treatment for filariasis, a repeat IVF cycle using the same stimulation protocol yielded a full-term pregnancy. This case is of particular importance because, to our knowledge, it is the first to describe a parasitic infection that probably affected the implantation of good-quality embryos in IVF cycles.

  6. HIGH INCIDENCE OF POLYSPERMIC FERTILIZATION IN BOVINE OOCYTES MATURED IN VITRO AFTER CRYOTOP VITRIFICATION.

    Science.gov (United States)

    Hwang, In-Sul; Kwon, Dae-Jin; Im, Gi-Sun; Tashima, Kazuya; Hochi, Shinichi; Hwang, Seongsoo

    2016-01-01

    Vitrification with the Cryotop device is the most promising technique for oocyte cryopreservation, but the high post-warming morphological survival of bovine oocytes does not guarantee high developmental competence after in vitro fertilization (IVF). This study was designed to examine achievement of normal fertilization in bovine oocytes vitrified-warmed with the Cryotop device. Oocytes were matured in vitro and vitrified-warmed after complete removal of the cumulus layers. Distribution of cortical granules (CGs) was assessed by Lens culinaris agglutinin (LCA) lectin staining. Ten hours after IVF, presumptive zygotes were analyzed for pronuclear formation. Day-8 blastocysts were harvested and stained with Hoechst-33342 for total cell counting. Both yield and mean cell number of the blastocysts were impaired by Cryotop vitrification. Incidence of polyspermic fertilization was three-times higher in vitrified oocytes compared to fresh oocytes. No difference in CG distribution was found between vitrified and fresh oocytes. Polyspermic fertilization induced in vitrified-warmed bovine oocytes may be one of the possible causes responsible for their low developmental potential.

  7. Fertilization of Mouse Gametes in Vitro Using a Digital Microfluidic System.

    Science.gov (United States)

    Huang, Hong-Yuan; Shen, Hsien-Hua; Chung, Lung-Yuan; Chung, Yu-Hsiang; Chen, Chih-Chen; Hsu, Chia-Hsien; Fan, Shih-Kang; Yao, Da-Jeng

    2015-12-01

    We demonstrated in vitro fertilization (IVF) using a digital microfluidic (DMF) system, so-called electrowetting on dielectric (EWOD). The DMF device was proved to be biocompatible and the DMF manipulation of a droplet was harmless to the embryos. This DMF platform was then used for the fertilization of mouse gametes in vitro and for embryo dynamic culture based on a dispersed droplet form. Development of the embryos was instantaneously recorded by a time-lapse microscope in an incubator. Our results indicated that increasing the number of sperms for IVF would raise the rate of fertilization. However, the excess of sperms in the 10 μL culture medium would more easily make the embryo dead during cell culture. Dynamic culture powered with EWOD can manipulate a single droplet containing mouse embryos and culture to the eight-cell stage. The fertilization rate of IVF demonstrated by DMF system was 34.8%, and about 25% inseminated embryos dynamically cultured on a DMF chip developed into an eight-cell stage. The results indicate that the DMF system has the potential for application in assisted reproductive technology.

  8. Effectiveness of in vitro fertilization in women with previous tubal sterilization.

    Science.gov (United States)

    Malacova, Eva; Kemp, Anna; Hart, Roger; Jama-Alol, Khadra; Preen, David B

    2015-03-01

    The objective was to determine the effectiveness of in vitro fertilization (IVF) on live-delivery rates in women who had previously undergone tubal sterilization. We examined first IVF live deliveries for women aged 20-44 years at their first embryo transfer (ET) with history of hospital admission for tubal sterilization in Western Australia (WA). The ET cycles (n=178) were ascertained over the period of 1996 to 2010 using WA hospital records. A control group of subfertile women matched by age was randomly selected (n=178). We used Kaplan-Meier curves and life-table analysis to evaluate the cumulative live-delivery rates. An overall cumulative live-delivery rate in women who had undergone previous tubal sterilization (31%) was comparable to that of subfertile controls (34%) within the first 24 months. Younger women (aged 20-34 years) with previous sterilization (34%) were slightly more likely to deliver an IVF live baby than older women (aged 35-39 and 40-44 years) (33% and 22%, respectively), although this difference was not statistically significant (p=.449). Women who desire fertility after a tubal sterilization procedure and undergo IVF have rates of pregnancy similar to age-matched subfertile IVF control patients. In vitro fertilization success in women who had undergone previous tubal sterilization is similar to that of the subfertile controls and thus does not depend on past fertility. Age is the most important predictive factor in achieving pregnancy. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Validation of a heterologous fertilization assay and comparison of fertilization rates of equine oocytes using in vitro fertilization, perivitelline, and intracytoplasmic sperm injections.

    Science.gov (United States)

    Sessions-Bresnahan, D R; Graham, J K; Carnevale, E M

    2014-07-15

    IVF in horses is rarely successful. One reason for this could be the failure of sperm to fully capacitate or exhibit hyperactive motility. We hypothesized that the zona pellucida (ZP) of equine oocytes prevents fertilization in vitro, and bypassing the ZP would increase fertilization rates. Limited availability of equine oocytes for research has necessitated the use of heterologous oocyte binding assays using bovine oocytes. We sought to validate an assay using bovine oocytes and equine sperm and then to demonstrate that bypassing the ZP using perivitelline sperm injections (PVIs) with equine sperm capacitated with dilauroyl phosphatidylcholine would result in higher fertilization rates than standard IVF in bovine and equine oocytes. In experiment 1, bovine oocytes were used for (1) IVF with bovine sperm, (2) IVF with equine sperm, and (3) intracytoplasmic sperm injections (ICSIs) with equine sperm. Presumptive zygotes were either stained with 4',6-diamidino-2-phenylindole from 18 to 26 hours at 2-hour intervals or evaluated for cleavage at 56 hours after addition of sperm. Equine sperm fertilized bovine oocytes; however, pronuclei formation was delayed compared with bovine sperm after IVF. The delayed pronuclear formation was not seen after ICSI. In experiment 2, bovine oocytes were assigned to the following five groups: (1) cumulus oocyte complexes (COCs) coincubated with bovine sperm; (2) COC exposed to sucrose then coincubated with bovine sperm; (3) COC coincubated with equine sperm; (4) COC exposed to sucrose, and coincubated with equine sperm; and (5) oocytes exposed to sucrose, and 10 to 15 equine sperm injected into the perivitelline (PV) space. Equine sperm tended (P = 0.08) to fertilize more bovine oocytes when injected into the PV space than after IVF. In experiment 3, oocytes were assigned to the following four groups: (1) IVF, equine, and bovine COC coincubated with equine sperm; (2) PVI of equine and bovine oocytes; (3) PVI with equine oocytes

  10. Quality management systems for your in vitro fertilization clinic's laboratory: Why bother?

    OpenAIRE

    Olofsson, Jan I; Banker, Manish R; Late Peter Sjoblom

    2013-01-01

    Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF) clinics and their laboratories. It is commonplace for Assisted Reproductive Technology (ART) laboratories to be required to have a quality control system. However, more effective Total Quality Management systems are now being implemented by an increasing number of ART clinics. In India, it is no...

  11. Maternal-foetal attachment during early pregnancy in Taiwanese women pregnant by in vitro fertilization.

    Science.gov (United States)

    Kuo, Pi-Chao; Bowers, Beverly; Chen, Yueh-Chih; Chen, Chung-Hey; Tzeng, Ya-Ling; Lee, Maw-Sheng

    2013-11-01

    The aim of this study was to investigate maternal-foetal attachment at 9, 12 and 20 weeks gestation and to identify factors that influenced maternal-foetal attachment in Taiwanese women who conceived by in vitro fertilization. Development of maternal-foetal attachment is an important part of taking on the maternal role. However, evidence about maternal-foetal attachment after assisted conception is inconclusive. A longitudinal design with repeated measures. A prospective, longitudinal design with repeated measures was used. Over an 18-month period in 2006-2008, a convenience sample of 160 women who conceived after undergoing successful in vitro fertilization were recruited from a major infertility care centre in Taiwan. Data were collected by self-reported measures, including: (1) Maternal-Foetal Attachment Scale; (2) Symptoms Checklist; (3) Pregnancy-related Anxiety Scale; (4) Social Support Apgar; (5) Chinese childbearing attitude Questionnaire; and (6) Awareness of Foetus Scale. The selected instruments to measure each variable were administered to participants at 9, 12 and 20 weeks gestation. Maternal-foetal attachment increased as pregnancy progressed from 9 to 20 weeks gestation. General linear mixed model showed predictors of maternal-foetal attachment included Chinese childbearing attitude, awareness of the foetus, and social support. Health provider awareness of cultural influences on the development of early maternal-foetal attachment of women pregnant by in vitro fertilization is needed. Prenatal education in early pregnancy might incorporate more information about foetal development to allow the mother to visualize her unborn child. Providing social support for women who were conceived by in vitro fertilization is beneficial to the development of maternal-foetal attachment. © 2013 Blackwell Publishing Ltd.

  12. Fertility of rabbit sperm exposed in vitro to cadmium and lead.

    Science.gov (United States)

    Foote, R H

    1999-01-01

    The heavy metals Cd2+ and Pb2+ have been associated with male reproductive toxicology, including possible inhibition of sperm undergoing hyperactivated motility, indicative of capacitation. The objective of the present study was to test fertility of rabbit sperm exposed to Cd2+ or Pb2+ in vitro, followed by insemination of superovulated does. Semen was washed to remove seminal plasma and minimize possible binding of the heavy metals by proteins. Only 400,000 treated or control sperm were inseminated as a sensitive test of treatment, and the time sperm resided in the female before possible fertilization was varied by inseminating from 0 to 12 h after ovulating the does. Only 6 of 22 males tested showed appreciable spontaneous hyperactivation and neither Cd2+ nor Pb2+ affected hyperactivation, or presumably associated capacitation. Sperm from four of these six males were used to inseminate 66 rabbits, and 1483 oocytes and embryos were collected about 27 h later. No effect of 0.1 mM Cd2+ on fertilizing ability of sperm was found (75% fertilization with control sperm and 78% with treated sperm). With 0.025 mM Pb2+ the fertilization rate in pregnant does only was 82% for controls and lower (68%) with treated sperm. These tested concentrations of Cd2+ and Pb2+ are much higher than reported concentrations in semen of exposed workers.

  13. Correlation of sperm penetration assay score with polyspermy rate in in-vitro fertilization.

    Science.gov (United States)

    Aoki, Vincent W; Peterson, C Matthew; Parker-Jones, Kirtly; Hatasaka, Harry H; Gibson, Mark; Huang, Ivan; Carrell, Douglas T

    2005-02-09

    BACKGROUND: The sperm penetration assay (SPA) is used to predict the fertilizing capacity of sperm. Thus, some programs rely on SPA scores to formulate insemination plans in conjunction with in-vitro fertilization (IVF) cycles. The purpose of this study was to evaluate if a relationship exists between SPA scores and polyspermy rates during conventional IVF cycles. METHODS: A total of 1350 consecutive IVF patients using conventional IVF insemination were evaluated in the study. Oocytes were inseminated three hours post-retrieval by the addition of 150,000 to 300,000 progressively motile sperm. Approximately 18 hours after insemination, the oocytes were evaluated for fertilization by the visualization of pronuclei. The presence of three or more pronuclei was indicative of polyspermy. Polyspermy rates, fertilization success, embryo quality, and pregnancy rates were analyzed retrospectively to evaluate their relationship with SPA score, count, motility, number of progressively motile sperm inseminated, oocyte pre-insemination incubation time, patient age, and diagnosis. RESULTS: A significant positive relationship was observed between SPA score and polyspermy rate (rs = 0.10, p polyspermy rates than those with abnormal SPA scores (6.3% +/- 1.5% vs. 2.0% +/- 0.7%, p polyspermy rates and IVF fertilization percentage. Additionally, there is a slight increase in clinical pregnancy rates, and embryo implantation rates with increased SPA. Furthermore, there is a slight decrease in spontaneous abortions rates related to increased SPA.

  14. Heterotopic triplet pregnancy complicated with ovarian hyperstimulation syndrome following in vitro fertilization.

    Science.gov (United States)

    Tomić, Vlatka; Tomić, Jozo; Kuna, Krunoslav; Zigmundovac-Klaić, Durda

    2011-12-01

    Heterotopic pregnancy is a rare event that occurs in less than 1% of pregnancies following in vitro fertilization and embryo transfer, especially when complicated with ovarian hyperstimulation syndrome. We report a case of a 31-year-old woman in the 6th gestational week of pregnancy achieved after in vitro fertilization, who was complaining of acute lower abdominal pain and distension, breathing difficulties and vaginal spotting. Transvaginal ultrasound examination and laboratory tests confirmed the ovarian hyperstimulation syndrome in the presence of two viable gestational sacs. The patient's condition worsened five days later with sudden onset of sharp abdominal pain, nausea and vomiting, along with impaired laboratory test values. Laparoscopy was attempted, but enlarged ovaries and adhesions prevented further procedure, which was then converted to mini-laparotomy. Operative removal of the right tubal pregnancy resulted in uncomplicated course of the intrauterine twin pregnancy and delivery of two healthy neonates by cesarean section at 37 weeks of gestation. Clinicians need to be aware of this rare complication where ovarian hyperstimulation syndrome coexists with heterotopic pregnancy after in vitro fertilization and embryo transfer procedure. Enlarged ovaries may mask accurate ultrasound diagnosis, but timely surgical intervention can prevent fatal consequences and result in normal course and outcome of intrauterine pregnancy.

  15. Expression of genes involved in lipid droplet formation (BSCL2, SNAP23 and COPA) during porcine in vitro adipogenesis.

    Science.gov (United States)

    Kociucka, Beata; Flisikowska, Tatiana; Mróz, Dariusz; Szczerbal, Izabela

    2016-11-01

    Adipogenesis is a complex process of fat cells development driven by the expression of numerous genes. Differentiation of progenitor cells into mature adipocytes is accompanied by changes in cell shape, as a result of lipid accumulation. In the present study, expression of three genes involved in lipid droplet formation (SNAP23, BSCL2 and COPA) was evaluated during porcine adipogenesis. It was found that mRNA levels of BSCL2 and SNAP23, but not COPA, increased during differentiation. Redistribution of SNAP23 protein to different cellular compartments was observed when comparing undifferentiated mesenchymal stem cells and differentiated adipocytes. The BSCL2 protein was found to be highly specific to cells with accumulated lipids, while COPA protein coated the lipid droplets. Obtained results indicated that the studied genes may be considered as candidates for fatness traits in pigs. Moreover, this study has shown that the porcine in vitro adipogenesis system provides a useful tool for the characterisation of novel genes involved in adipose tissue accumulation.

  16. Staining and in vitro toxicity of dithizone with canine, porcine, and bovine islets.

    Science.gov (United States)

    Clark, S A; Borland, K M; Sherman, S D; Rusack, T C; Chick, W L

    1994-01-01

    Dithizone (DTZ) is a recognized diabetogenic agent in vivo, and a supravital stain commonly used for identification of islets to be used for transplantation. In the present studies, we compared DTZ staining of freshly isolated and cultured canine, bovine, and porcine islets, and the effect of DTZ on the function and viability of islets. Incubation with DTZ resulted in staining of canine and porcine islets, but no discernible staining with bovine islets. Insulin content of porcine, canine, and bovine islet was 2.0 +/- 0.2, 2.2 +/- 0.3, and 1.9 +/- 0.2 mU/EIN, indicating a lack of correspondence of DTZ staining and insulin content. Seven days of culture with canine islets resulted in > or = 50% reduction of DTZ stained cells. Exposure to DTZ at 50 micrograms/mL resulted in a maximal number of stained cells in preparations of purified islets (80-85%; counted after dispersion), a lower percentage of cells stained faintly at 20 micrograms/mL (50-55%), with no discernible staining at 10 micrograms/mL. Prolonged exposure of islets (4-48 h) to 20 micrograms/mL DTZ led to reduced insulin secretion and islet cell death. Incubation of canine or porcine islets with 100 micrograms/mL of DTZ for 0.5 h resulted in a dramatic loss of viability and diminished insulin secretory function, which was not reversed with continued culture. The concentration dependence of toxic effects paralleled the concentration dependence of cellular staining. The minimally effective staining concentration (20 micrograms/mL) also resulted in a loss of viability. An additional assessment of DTZ toxicity was made using the RIN-38 beta-cell line, which shows no discernible staining with DTZ.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Urinary bisphenol A concentrations and association with in vitro fertilization outcomes among women from a fertility clinic.

    Science.gov (United States)

    Mínguez-Alarcón, Lidia; Gaskins, Audrey J; Chiu, Yu-Han; Williams, Paige L; Ehrlich, Shelley; Chavarro, Jorge E; Petrozza, John C; Ford, Jennifer B; Calafat, Antonia M; Hauser, Russ

    2015-09-01

    Are urinary BPA concentrations associated with in vitro fertilization (IVF) outcomes among women attending an academic fertility center? Urinary BPA concentrations were not associated with adverse reproductive and pregnancy outcomes among women from a fertility clinic. Bisphenol A (BPA), an endocrine disruptor, is detected in the urine of most Americans. Although animal studies have demonstrated that BPA reduces female fertility through effects on the ovarian follicle and uterus, data from human populations are scarce and equivocal. This prospective cohort study between 2004 and 2012 at the Massachusetts General Hospital Fertility Center included 256 women (n = 375 IVF cycles) who provided up to two urine samples prior to oocyte retrieval (total N = 673). Study participants were women enrolled in the Environment and Reproductive Health (EARTH) Study. Intermediate and clinical end-points of IVF treatments were abstracted from electronic medical records. We used generalized linear mixed models with random intercepts to evaluate the association between urinary BPA concentrations and IVF outcomes adjusted by age, race, body mass index, smoking status and infertility diagnosis. The specific gravity-adjusted geometric mean of BPA was 1.87 µg/l, which is comparable to that for female participants in the National Health and Nutrition Examination Survey, 2011-2012. Urinary BPA concentrations were not associated with endometrial wall thickness, peak estradiol levels, proportion of high quality embryos or fertilization rates. Furthermore, there were no associations between urinary BPA concentrations and implantation, clinical pregnancy or live birth rates per initiated cycle or per embryo transfer. Although we did not find any associations between urinary BPA concentrations and IVF outcomes, the relation between BPA and endometrial wall thickness was modified by age. Younger women (women (≥37 years old) had thinner endometrial thickness across increasing quartiles of

  18. Role of ataxia-telangiectasia mutated (ATM) in porcine oocyte in vitro maturation.

    Science.gov (United States)

    Lin, Zi-Li; Kim, Nam-Hyung

    2015-06-01

    Ataxia-telangiectasia mutated (ATM) is critical for the DNA damage response, cell cycle checkpoints, and apoptosis. Significant effort has focused on elucidating the relationship between ATM and other nuclear signal transducers; however, little is known about the connection between ATM and oocyte meiotic maturation. We investigated the function of ATM in porcine oocytes. ATM was expressed at all stages of oocyte maturation and localized predominantly in the nucleus. Furthermore, the ATM-specific inhibitor KU-55933 blocked porcine oocyte maturation, reducing the percentages of oocytes that underwent germinal vesicle breakdown (GVBD) and first polar body extrusion. KU-55933 also decreased the expression of DNA damage-related genes (breast cancer 1, budding uninhibited by benzimidazoles 1, and P53) and reduced the mRNA and protein levels of AKT and other cell cycle-regulated genes that are predominantly expressed during G2/M phase, including bone morphogenetic protein 15, growth differentiation factor 9, cell division cycle protein 2, cyclinB1, and AKT. KU-55933 treatment decreased the developmental potential of blastocysts following parthenogenetic activation and increased the level of apoptosis. Together, these data suggested that ATM influenced the meiotic and cytoplasmic maturation of porcine oocytes, potentially by decreasing their sensitivity to DNA strand breaks, stimulating the AKT pathway, and/or altering the expression of other maternal genes. © 2015 International Federation for Cell Biology.

  19. Effect of high and low antral follicle count in pubertal beef heifers on in vitro fertilization (IVF)

    Science.gov (United States)

    Pubertal heifers can be classified between those with high (= 25) and low (= 15) antral follicle counts (AFC). The objective of this study was to determine oocyte development and maturation (e.g., fertility) in an in vitro fertilization (IVF) system for high and low AFC heifers. From a pool of 120...

  20. Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells

    DEFF Research Database (Denmark)

    Secher, Jan Ole Bertelsen; Ceylan, Ahmet; Mazzoni, Gianluca

    2017-01-01

    Derivation and stable maintenance of porcine induced pluripotent stem cells (piPSCs) is challenging. We herein systematically analyzed two piPSC lines, derived by lentiviral transduction and cultured under either leukemia inhibitory factor (LIF) or fibroblast growth factor (FGF) conditions, to shed...... more light on the underlying biological mechanisms of porcine pluripotency. LIF-derived piPSCs were more successful than their FGF-derived counterparts in the generation of in vitro chimeras and in teratoma formation. When LIF piPSCs chimeras were transferred into surrogate sows and allowed to develop...

  1. The significance of fertile pigs vaccination against porcine parvovirus infection in the prevention of intrauterine infection and formation of colostrum piglets immunity

    Directory of Open Access Journals (Sweden)

    Stojanac Nenad

    2014-01-01

    Full Text Available The aim of this study was to come closer to the knowledge of piglets protection during intrauterine life, as well as formation of colostrum immunity against Porcine Parvovirus Infection (PPV, on the basis of detailed antibody titer analysis from the blood serum of pigs 7 days before previous weaning, 30 days before mating and their piglets during first 3 days of life. The research included 60 fertile pigs and 300 of their offspring. For that purpose we have examined antibody titer specific for PPV in blood serum of vaccinated fertile pigs on 70th and 113th day of gestation, and in the blood serum of piglets originated from itemized fertile pigs during first day of life, before colostrums consummation and also during 3rd day of life. On the 70th day of gestation, in the fertile pigs blood serum, average antibody titer specific for PPV, value of 12.60 was determined, what represents adequate level for solid protection against PPV infection. This was confirmed undoubtedly by examination results of antibodies in the blood serum of piglets before colostrum consummation, which was 100% negative. Titer drop in the blood serum of fertile pigs, on 113th day of gestation (on the level of 8.7 came as a result of specific antibodies transfer from the mother’s blood flow to the colostrum. The above-mentioned is supported by the fact that on the 3rd day of life there was confirmed high average body titer level (13.37 in newborn piglets body serum has been confirmed. The principle of fertile pigs vaccination 7 days before weaning and one month before gilts insemination is an efficient measure which prevents intrauterine infection occurrence during the entire gestation process. The level of antibodies specific for PPV which is determined in gilts blood serum after vaccination can be a result of both primary and secondary immune response (animal infected before first vaccination, as well as absence of gilts revaccination, what is usually recommended by

  2. Phospholipase C-zeta deficiency as a cause for repetitive oocyte fertilization failure during ovarian stimulation for in vitro fertilization with ICSI: a case report.

    Science.gov (United States)

    Chithiwala, Zahabiya H; Lee, Hoi Chang; Hill, David L; Jellerette-Nolan, Teru; Fissore, Rafael; Grow, Daniel; Dumesic, Daniel A

    2015-09-01

    The purpose of this study is to describe impaired oocyte fertilization from phospholipase C-zeta (PLC-ζ) deficiency in normal-appearing sperm that was successfully treated using calcium (Ca(2+)) ionophore with intracytoplasmic sperm injection (ICSI) of oocytes matured in vitro. An infertile couple undergoing in vitro fertilization (IVF) experienced failed oocyte fertilization following ICSI with normal-appearing sperm. A semen sample collected from the patient was used to assess the expression of sperm PLC- ζ protein by Western blot analysis and immunofluorescence and PLC-ζ bioactivity by an in vitro model of Ca(2+) release. A second IVF cycle was performed using Ca(2+) ionophore with ICSI to enhance Ca(2+)-induced oocyte activation of oocytes matured in vitro. Sperm PLC-ζ protein deficiency was demonstrated by Western blot analysis and immunofluorescence and confirmed by reduced PLC-ζ bioactivity using an in vitro model of Ca(2+) release. Nevertheless, with this sperm and supplementation of Ca(2+) ionophore following ICSI, fertilization of four of six oocytes matured in vitro was obtained. In addition, four embryos underwent cleavage and two of them reached the blastocyst stage. Transfer of these blastocysts into the uterus led to a single pregnancy and live birth. Deficiency of PLC-ζ in normal-appearing human sperm is associated with impaired Ca(2+)-dependent oocyte activation during ICSI. Under this condition, use of Ca(2+) ionophore following ICSI of oocytes matured in vitro improves embryo developmental competence, possibly through the activation of Ca(2+)-dependent mechanisms governing fertilization and preimplantation embryogenesis.

  3. Hyperactivation of stallion sperm is required for successful in vitro fertilization of equine oocytes.

    Science.gov (United States)

    McPartlin, L A; Suarez, S S; Czaya, C A; Hinrichs, K; Bedford-Guaus, S J

    2009-07-01

    Capacitation is a complex and not well-understood process that encompasses all the molecular changes sperm must undergo to successfully fertilize an oocyte. In vitro fertilization has remained elusive in the horse, as evidenced by low in vitro fertilization (IVF) rates (0%-33%); moreover, only two foals have ever been produced using IVF. Incubation of stallion sperm in modified Whittens supplemented with bovine serum albumin and sodium bicarbonate yielded significant rates of time-dependent protein tyrosine phosphorylation and induced acrosomal exocytosis, consistent with capacitation. The objective of this study was to characterize stallion sperm hyperactivation and to test whether hyperactivation of capacitated sperm supported equine IVF. Treatment of sperm with procaine, an anesthetic shown to induce hyperactivation in other mammalian species, resulted in the decrease of three motility variables indicative of hyperactivation: straight line velocity (P = 0.029), straightness (P = 0.001), and linearity (P = 0.002). We demonstrated that procaine-induced hyperactivation was not regulated by changes in protein tyrosine phosphorylation and that it did not induce acrosomal exocytosis in capacitated sperm compared with calcium ionophore (P > 0.05), similar to findings in the bovine. Most notably, by coupling our capacitating conditions with the induction of hyperactivation using procaine, we have achieved the novel result of substantial and reproducible percentages of fertilized mare oocytes (60.7%) in our IVF experiments. Conversely, sperm incubated in capacitating conditions but not treated with procaine did not fertilize (0%). These results support the hypothesis that capacitation and hyperactivation are required for successful IVF in the equine.

  4. Kisspeptin-54 triggers egg maturation in women undergoing in vitro fertilization.

    Science.gov (United States)

    Jayasena, Channa N; Abbara, Ali; Comninos, Alexander N; Nijher, Gurjinder M K; Christopoulos, Georgios; Narayanaswamy, Shakunthala; Izzi-Engbeaya, Chioma; Sridharan, Mathini; Mason, Alexina J; Warwick, Jane; Ashby, Deborah; Ghatei, Mohammad A; Bloom, Stephen R; Carby, Anna; Trew, Geoffrey H; Dhillo, Waljit S

    2014-08-01

    Patients with mutations that inactivate kisspeptin signaling are infertile. Kisspeptin-54, the major circulating isoform of kisspeptin in humans, potently stimulates reproductive hormone secretion in humans. Animal studies suggest that kisspeptin is involved in generation of the luteinizing hormone surge, which is required for ovulation; therefore, we hypothesized that kisspeptin-54 could be used to trigger egg maturation in women undergoing in vitro fertilization therapy. Following superovulation with recombinant follicle-stimulating hormone and administration of gonadotropin-releasing hormone antagonist to prevent premature ovulation, 53 women were administered a single subcutaneous injection of kisspeptin-54 (1.6 nmol/kg, n = 2; 3.2 nmol/kg, n = 3; 6.4 nmol/kg, n = 24; 12.8 nmol/kg, n = 24) to induce a luteinizing hormone surge and egg maturation. Eggs were retrieved transvaginally 36 hours after kisspeptin injection, assessed for maturation (primary outcome), and fertilized by intracytoplasmic sperm injection with subsequent transfer of one or two embryos. Egg maturation was observed in response to each tested dose of kisspeptin-54, and the mean number of mature eggs per patient generally increased in a dose-dependent manner. Fertilization of eggs and transfer of embryos to the uterus occurred in 92% (49/53) of kisspeptin-54-treated patients. Biochemical and clinical pregnancy rates were 40% (21/53) and 23% (12/53), respectively. This study demonstrates that a single injection of kisspeptin-54 can induce egg maturation in women with subfertility undergoing in vitro fertilization therapy. Subsequent fertilization of eggs matured following kisspeptin-54 administration and transfer of resulting embryos can lead to successful human pregnancy. ClinicalTrials.gov NCT01667406.

  5. [First experience with an in vitro fertilization program at the Rostock University Clinic of Gynecology and Obstetrics].

    Science.gov (United States)

    Sudik, R; Fliess, F R

    1984-01-01

    In vitro fertilization as an aid in diagnosis and treatment of the sterile couple has been used at the university clinic of gynecology and obstetrics in Rostock since the beginning of 1983. 17 women were treated in the first series of in vitro-fertilization and embryo transfer in September and October 1983. After treatment with clomiphene and HCG in 16 women one or more oocytes were harvested. In 14 women one or more oocytes were fertilized. Embryo transfer was performed in five cases. No pregnancy resulted. The causes of the low cleavage rate and possibilities for improvement of the results are discussed.

  6. In vitro fertilization and embryo culture strongly impact the placental transcriptome in the mouse model.

    Directory of Open Access Journals (Sweden)

    Patricia Fauque

    Full Text Available BACKGROUND: Assisted Reproductive Technologies (ART are increasingly used in humans; however, their impact is now questioned. At blastocyst stage, the trophectoderm is directly in contact with an artificial medium environment, which can impact placental development. This study was designed to carry out an in-depth analysis of the placental transcriptome after ART in mice. METHODOLOGY/PRINCIPAL FINDINGS: Blastocysts were transferred either (1 after in vivo fertilization and development (control group or (2 after in vitro fertilization and embryo culture. Placentas were then analyzed at E10.5. Six percent of transcripts were altered at the two-fold threshold in placentas of manipulated embryos, 2/3 of transcripts being down-regulated. Strikingly, the X-chromosome harbors 11% of altered genes, 2/3 being induced. Imprinted genes were modified similarly to the X. Promoter composition analysis indicates that FOXA transcription factors may be involved in the transcriptional deregulations. CONCLUSIONS: For the first time, our study shows that in vitro fertilization associated with embryo culture strongly modify the placental expression profile, long after embryo manipulations, meaning that the stress of artificial environment is memorized after implantation. Expression of X and imprinted genes is also greatly modulated probably to adapt to adverse conditions. Our results highlight the importance of studying human placentas from ART.

  7. Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification.

    Science.gov (United States)

    Morató, Roser; Castillo-Martín, Míriam; Yeste, Marc; Bonet, Sergi

    2016-06-01

    The aim of our study was to assess whether the cryotolerance of in vitro-produced embryos could be influenced by the length of in vitro culture and size of blastocoel cavity before vitrification, using the pig as a model. For this purpose we analysed the cryoresistance and apoptosis rate of blastocysts at different stages of development as derived on Day 5 and 6 of in vitro culture. Blastocysts were subsequently vitrified, warmed and cultured for 24h. Re-expansion rates were recorded at 3 and 24h and total cell number and apoptotic cells were determined at 24h. Day-6 blastocysts showed the highest rates of survival after warming, which indicates higher quality compared with Day-5 blastocysts. Higher re-expansion rates were observed for expanded blastocysts and those in the process of hatching when compared with early blastocysts. Total cell number and apoptotic cells were affected by blastocyst stage, vitrification-warming procedures and length of in vitro culture, as expanding and hatching-hatched blastocysts from Day 6 presented higher percentages of apoptotic cells than fresh blastocysts and blastocysts vitrified at Day 5. Our findings suggest that the cryotop vitrification method is useful for the cryopreservation of porcine blastocysts presenting a high degree of expansion, particularly when vitrification is performed after 6 days of in vitro culture. Furthermore, these results show that faster embryo development underlies higher blastocyst cryotolerance and provide evidence that blastocoel cavity expansion before vitrification is a reliable index of in vitro-produced embryo quality and developmental potential.

  8. Effects of difructose dianhydride (DFA)-IV on in vitro fertilization in pigs.

    Science.gov (United States)

    Yi, Young-Joo; Kamala-Kannan, S; Lim, Jeong-Muk; Oh, Byung-Taek; Lee, Sang-Myeong

    2017-09-26

    Difructose dianhydride IV (DFA-IV) is produced from levan, which is a natural polysaccharide that belongs to the fructan family, through the activity of levan fructotransferase (LF) derived from microorganisms. Recently, DFA-IV has been expected to have diverse applications in the food and medical industry. Here, we examined the potential application of DFA-IV forin vitro fertilization (IVF) in pigs. In the assessment of acrosomal integrity during incubation, intact acrosomal or viable spermatozoa were highly sustained in 0.1% or 0.25% DFA-IV (69.8%-70.8%,PDFA-IV, and 0.1%-0.5% DFA-IV in particular significantly decreased ROS production relative to that seen with no addition or 0.75% DFA-IV. Total fertilization (mono+ polyspermic oocyte) rate was significantly higher in the addition of 0.1% DFA-IV (94.2%) than with other concentrations (71.8%-86.7%,PDFA-IV significantly increased the fertilization rate (PDFA-IV exhibited higher embryonic development and blastocyst formation than those treated with other concentrations (PDFA-IV during IVF improved fertilization and embryonic development, suggesting the possible use of novel sugars for enhancement of assisted reproductive technology (ART) in mammals.

  9. Oxygen tension and oocyte density during in vitro maturation affect the in vitro fertilization of bovine oocytes

    Directory of Open Access Journals (Sweden)

    Angelo Bertani Giotto

    2015-12-01

    Full Text Available Oocyte maturation is the key factor affecting the fertilization and embryonic development. Factors such as oocyte density and oxygen tension can directly influence the IMV. Thus, the objective of this study was to evaluate the effect of the association of oxygen tensions (5% or 20% with different oocyte densities (1:10?l or 1:20?l in the in vitro maturation (IVM of bovine oocytes on maturation and fertilization rates, ROS production and antioxidant activity. Three experiments were performed with bovine oocytes that were obtained from slaughterhouse ovaries. After selection, the oocytes were randomly distributed in four treatments: 1:10/5%; 1:10/20%; 1:20/5%and 1:20/20% for each experiment. In experiment I, nuclear maturation status and cytoplasmic maturation were evaluated through detection of the first polar body by immunofluorescence and the mitochondrial reorganization assay. In experiment II, ROS production and antioxidant activity were analyzed in oocytes and IVM medium after 24 h of maturation through detection of ROS, reduced glutathione (GSH and Superoxide dismutase activity by spectrofluorimetric methods. In experiment III, fertilization was evaluated through pronucleus formation, sperm penetration with or without decondensation and polyspermy rates by immunofluorescence. In experiment I, the nuclear maturation and cytoplasmic maturation were similar among treatments (P>0.05. In experiment II, reactive oxygen species in oocytes were elevated in treatments with low oxygen tension which was independent of oocyte density (P<0.05. Additionally, ROS levels in IVM medium were higher in treatments with high oocyte density by volume of medium, which was independent of oxygen tension (P<0.05. In Experiment III, the fertilization and penetration rates were higher in the treatment with 20% oxygen tension and high oocyte density (P<0.05. Furthermore, a high incidence of polyspermy was observed in groups with high oxygen tension and low oocyte

  10. Chemical composition and in vitro digestibility of whole-crop maize fertilized with synthetic fertilizer or digestate and harvested at two maturity stages in Boreal growing conditions

    Directory of Open Access Journals (Sweden)

    Mahmoud F. Seleiman

    2017-04-01

    Full Text Available Maize cultivation for silage could be a sustainable option in Boreal conditions, especially when combined with nutrient recycling. Effects of digestate (sludge from biogas of domestic origin application in comparison with synthetic fertilizer and two maturity stages on chemical composition and in vitro digestibility of whole-crop maize were investigated. Starch, neutral detergent fiber, water soluble carbohydrate (WSC and digestible organic matter (DOM contents of maize did not differ in response to the two fertilizer treatments. However, starch, DOM and metabolizable energy of maize increased, while ash, crude protein and WSC contents decreased with increasing maize maturity. Heavy metals in maize fertilized with digestate remained low. The results indicate that whole-crop maize fertilized with digestate and harvested at 150 days after sowing is a promising feed and has good nutritive value, even in Boreal conditions.

  11. Controlled ovarian stimulation and triggers in in vitro fertilization: protocol personalization key to optimize outcomes.

    Science.gov (United States)

    Maher, Jacqueline Y; Christianson, Mindy S

    2017-03-21

    The past four decades have delivered many advancements to improve in vitro fertilization (IVF) outcomes. These include a delicate balance of controlled ovarian hyperstimulation (COH) without causing ovarian hyperstimulation syndrome (OHSS), a safe oocyte retrieval, fertilization and embryo culture, endometrial growth and receptivity to promote implantation, and luteal support to maintain the pregnancy. Contemporary IVF practice includes both the classic COH protocols as well as protocols for poor responders and those for specific patient populations. An assortment of agents have been developed and utilized in various combinations to improve COH outcomes and promote oocyte maturation while decreasing the risk of OHSS. Various protocols have evolved over time. Ideal practices involve selecting the optimal protocol for a personalized, patient-specific stimulation and trigger.

  12. Quality management systems for your in vitro fertilization clinic′s laboratory: Why bother?

    Directory of Open Access Journals (Sweden)

    Jan I Olofsson

    2013-01-01

    Full Text Available Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF clinics and their laboratories. It is commonplace for Assisted Reproductive Technology (ART laboratories to be required to have a quality control system. However, more effective Total Quality Management systems are now being implemented by an increasing number of ART clinics. In India, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services. This review contains the proceedings a quality management session at the Indian Fertility Experts Meet (IFEM 2010 and focuses on the creation of a patient-oriented best-in-class IVF laboratory.

  13. Quality management systems for your in vitro fertilization clinic's laboratory: Why bother?

    Science.gov (United States)

    Olofsson, Jan I; Banker, Manish R; Sjoblom, Late Peter

    2013-01-01

    Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF) clinics and their laboratories. It is commonplace for Assisted Reproductive Technology (ART) laboratories to be required to have a quality control system. However, more effective Total Quality Management systems are now being implemented by an increasing number of ART clinics. In India, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services. This review contains the proceedings a quality management session at the Indian Fertility Experts Meet (IFEM) 2010 and focuses on the creation of a patient-oriented best-in-class IVF laboratory.

  14. Chromosomal preparations of human triploid zygotes and embryos fertilized in vitro.

    Science.gov (United States)

    Macas, E; Suchanek, E; Grizelj, V; Puharic, I; Simunic, V

    1988-12-01

    Forty-eight zygotes with more than two pronuclei were identified after in vitro fertilization, representing 6.1% of all fertilized oocytes. The chromosome preparations from pronuclear stage to the cleaved human embryos were examined. Prophase was found in eight out of ten zygotes. The spreading of chromosomes allowed an adequate counting in only two cases. Six of the eight preparations displayed a late prophase. In this stage each haploid group of chromosomes can be analysed separately. Kariogamy usually occurred 4 to 5 h after the pronuclei had disappeared, and polyploid number of chromosomes were found in well-spread metaphases. The chromosomal preparations were made for eleven human embryos arising from zygotes with three pronuclei. Out of ten preparations, where the chromosomes could be counted, seven embryos (70%) contained hypodiploidic groups of chromosomes. In two of the cases, however, triploid metaphases were found, and in the last one a triploid/diploid mosaicism.

  15. Hsp70 positively regulates porcine circovirus type 2 replication in vitro.

    Science.gov (United States)

    Liu, Jie; Bai, Juan; Zhang, Lili; Jiang, Zhihua; Wang, Xianwei; Li, Yufeng; Jiang, Ping

    2013-12-01

    The Hsp70 chaperone plays a central role in multiple processes within cells. Porcine circovirus type 2 (PCV2) is the essential causal agent of post-weaning multisystemic wasting syndrome (PMWS), which has spread worldwide. But the mechanism of PCV2 replication remains poorly understood. In this study, we firstly found the positive effect of heat stress on the replication of PCV2 in the continuous porcine monocytic cell line 3D4/31. Downregulation of Hsp70 by the specific chaperone inhibitor Quercetin or RNA interference and upregulation of Hsp70 by expression from a recombinant adenovirus showed that Hsp70 enhanced PCV2 genome replication and virion production. A specific interaction between Hsp70 and PCV2 Cap was confirmed by colocalization by confocal microscopy and co-immunoprecipitation. Furthermore, the NF-κB pathway was activated and caspase-3 activity was reduced when Hsp70 was overexpressed in PCV2-infected 3D4/31 cells. These data suggested that Hsp70 positively regulated PCV2 replication, which being helpful for understanding the molecular mechanism of PCV2 infection. © 2013 Elsevier Inc. All rights reserved.

  16. In vitro embryo development in the pig: impact of oocyte maturation milieu on blastocyst morphology and viability

    NARCIS (Netherlands)

    Kidson, Annadie

    2004-01-01

    In this study, porcine embryos were produced in vitro from slaughterhouse sow or gilt oocytes which were matured and fertilized in vitro and subsequently cultured to the blastocyst stage. In vitro produced blastocysts are of poorer quality than their in vivo counterparts, and suffer from a high

  17. Effect of heat stress on development in vitro and in vivo and on synthesis of heat shock proteins in porcine embryos.

    Science.gov (United States)

    Kojima, T; Udagawa, K; Onishi, A; Iwahashi, H; Komatsu, Y

    1996-04-01

    The present study was conducted (1) to examine the effect of an acute increase in ambient temperature on the development of porcine day 6 embryos in culture and after transfer to recipient gilts, and (2) to analyze intracellular production of heat shock proteins (hsps). The viability of porcine day 6 embryos following a temporary acute elevation in ambient temperature (at 42 degrees-45.5 degrees C and for 10-180 min) was examined. Synthesis of 70 kDa hsp (hsp70) and 90 kDa hsp (hsp90) was determined by SDS-PAGE and Western blot analysis in porcine day 6 embryos subjected to heat stresses. Nonheat-stressed embryos were considered as control. Significantly higher numbers of viable nuclei were observed in treatment groups of 42 degrees C-10 min (236.6 +/- 71.4; P heat stress compared to control (82.5 +/- 47.3 microns), while heat stress with 43 degrees C for > or = 60 min, 44 degrees-44.5 degrees C for > or = 30 min, or 45 degrees-45.5 degrees C for > or = 10 min impaired their survival, as assessed by differences in number of viable nuclei. The embryos subjected to heat stresses under the conditions of 42 degrees C-180 min, 43 degrees C-10 min, 43 degrees C-30 min, 44 degrees C-10 min, or 45 degrees C-10 min developed to normal piglets after transfer to recipient gilts. Overall pregnancy rate was 75% (6/8), and farrowing rate 62.5% (5/8). Of heat-stressed embryos transferred, 59% (36/61) developed to normal piglets. Heat-stress conditions of 42 degrees C for 180 min, 43 degrees C for 30 min, 44 degrees C for 10 min, and 45 degrees C for 10 min were determined as critical with respect to the in vitro and in vivo survival of porcine embryos. Porcine day 6 embryos constitutively synthesized hsp70 even without heat stress, while hsp90 was detected only at trace level. Neither hsp70 nor hsp90 levels increased in the embryos subjected to heat stresses. In conclusion, porcine day 6 embryos could continue to develop in vivo or during in vitro culture after exposure to acute

  18. THE EFFECT OF NITROGEN FERTILIZATION OF MAIZE ON PROTEIN CONCENTRATION AND IN VITRO FEMENTABILITY OF GRAIN

    Directory of Open Access Journals (Sweden)

    D BABNIK

    2002-12-01

    Full Text Available The effect of nitrogen fertilization of maize on fermentability of maize grain in the rumen was studied by means of in vitro method based on the measurement of gas produced during the incubation of samples with rumen liquor. Gas production was recorded continuously up to 72 h incubation time and cumulative gas production was described by the Gompertz equation Y=A*exp(-exp(-d*(t-tm. Seven treatments, one of them unfertilized and others fertilized with 100 to 250 kg N ha–1, were compared. Grain yield and concentration of crude protein (CP in grain increased linearly with nitrogen fertilization. Grain yield increased for 25 kg dry matter (DM ha–1 and CP concentration for 0.13 g kg–1 DM per each additional kg of N. Concentration of CP in grain, which varied from 83 to 115 g kg–1 DM, was closely related to the dynamics of gas production. The maximal gas production rate (MPR was negatively related to CP concentration in the grain (R2 = 0.53; p < 0.10 and the time of MPR (tm was positively related to the amount of added N (R2 = 0.74; p < 0.05 and concentration of CP in the grain (R2 = 0.88; p < 0.01. It is likely that intensive N fertilization of maize limits ruminal digestion of maize starch. Due to the shift of starch digestion from the rumen to lower gastrointestinal tract better utilization of energy can be expected in maize grain of extensively fertilized maize than in the grain of maize, in which supply of N is sub-optimal.

  19. Male smokers have a decreased success rate for in vitro fertilization and intracytoplasmic sperm injection.

    Science.gov (United States)

    Zitzmann, Michael; Rolf, Claus; Nordhoff, Verena; Schräder, Guido; Rickert-Föhring, Melanie; Gassner, Paul; Behre, Hermann M; Greb, Robert R; Kiesel, Ludwig; Nieschlag, Eberhard

    2003-06-01

    Smoking by one or both partners can adversely affect IVF outcome. We investigated whether smoking may also play a role in the success rate of intracytoplasmic sperm injection (ICSI), in which initial steps of fertilization are bypassed. Three hundred one couples (ICSI: 153, IVF: 148) participated in 415 treatment cycles (ICSI: 202, IVF: 213). One hundred thirty-nine men were habitual smokers (ICSI: 71, IVF: 68). Seventy-seven women were smokers (ICSI: 41, IVF: 36). Multiple nominal regression analyses of various steps of assisted reproduction included smoking status, age, semen parameters, and number of embryos transferred. Reproductive and andrology unit of the university. Three hundred one couples seeking fertility treatment. Assisted reproduction by in vitro fertilization (IVF) or ICSI. Clinical pregnancy. Intracytoplasmic sperm injection success (clinical pregnancy) in women with smoking male partners was 22% and was 38% with nonsmoking partners. Similar results were seen for IVF, with 18% vs. 32%. Multinominal logistic regression analysis revealed smoking in men to be a significant predictor of ICSI outcome, along with female age and the number of embryos transferred, whereas clinical pregnancies after IVF were dependent on smoking in men, number of embryos transferred, sperm motility, and female age. Female smoking influenced the number of oocytes retrieved and the fertilization rate of oocytes in IVF but not in ICSI. The odds ratio for failure of ICSI for male smokers in comparison to male nonsmokers was 2.95 (IVF: 2.65). Smoking by males decreases the success rates of assisted reproduction procedures, not only in IVF, but also in ICSI. Apart from putative adverse effects during fertilization, altered DNA in spermatozoa might hamper development of the embryo.

  20. Urinary Concentrations of Organophosphate Flame Retardant Metabolites and Pregnancy Outcomes among Women Undergoingin VitroFertilization.

    Science.gov (United States)

    Carignan, Courtney C; Mínguez-Alarcón, Lidia; Butt, Craig M; Williams, Paige L; Meeker, John D; Stapleton, Heather M; Toth, Thomas L; Ford, Jennifer B; Hauser, Russ

    2017-08-25

    Evidence from animal studies suggests that exposure to organophosphate flame retardants (PFRs) can disrupt endocrine function and impair embryo development. However, no epidemiologic studies have been conducted to evaluate effects on fertility and pregnancy outcomes. We evaluated associations between urinary concentrations of PFR metabolites and outcomes of in vitro fertilization (IVF) treatment among couples recruited from an academic fertility clinic. This analysis included 211 women enrolled in the Environment And Reproductive Health (EARTH) prospective cohort study (2005-2015) who provided one or two urine samples per IVF cycle. We measured five urinary PFR metabolites [bis(1,3-dichloro-2-propyl) phosphate (BDCIPP), diphenyl phosphate (DPHP), isopropylphenyl phenyl phosphate (ip-PPP), tert-butylphenyl phenyl phosphate (tb-PPP), and bis(1-chloro-2-propyl) phosphate (BCIPP)] using negative electrospray ionization liquid chromatography tandem mass spectrometry (LC-MS/MS). Molar concentrations of the urinary PFR metabolites were summed. We used multivariable generalized linear mixed models to evaluate the association of the PFR metabolites with IVF outcomes, accounting for multiple IVF cycles per woman. Detection frequencies were high for BDCIPP (87%), DPHP (94%), and ip-PPP (80%), but low for tb-PPP (14%) and BCIPP (0%). We observed decreased success for several IVF outcomes across increasing quartiles of both summed and individual PFR metabolites (DPHP and ip-PPP) in our adjusted multivariable models. Significant declines in adjusted means from the lowest to highest quartile of ΣPFR were observed for the proportion of cycles resulting in successful fertilization (10% decrease), implantation (31%), clinical pregnancy (41%), and live birth (38%). Using IVF to investigate human reproduction and pregnancy outcomes, we found that concentrations of some urinary PFR metabolites were negatively associated with proportions of successful fertilization, implantation

  1. Incubation of boar spermatozoa in viscous media by addition of methylcellulose improves sperm quality and penetration rates during in vitro fertilization.

    Science.gov (United States)

    González-Abreu, David; García-Martínez, Soledad; Fernández-Espín, Vanesa; Romar, Raquel; Gadea, Joaquín

    2017-04-01

    This work was designed to study whether viscous media can improve the in vitro sperm functionality in pigs by using methylcellulose as a thickener. Viscosity of porcine oviductal fluid (POF) was compared with culture medium (Tyrode's) supplemented with methylcellulose (MET 0, 0.5 and 1% w/v). Spermatozoa were incubated in the different media (0, 1 and 2 h) and sperm motion parameters, lipid membrane disorder, plasma membrane integrity and reactive oxygen species (ROS) formation were assessed. Fertilization results were assessed i) preincubating spermatozoa in the viscous media followed by gamete coculture in a non-viscous medium; and ii) gamete coculture in the viscous media. Viscosity of POF from early luteal phase was higher than late follicular phase. Medium without methylcellulose presented constant viscosity with increased shear rate, while viscosity of the POF and media with methylcellulose was reduced by increased shear rates. Methylcellulose improved sperm linearity, straightness and the proportion of fast-linear spermatozoa. Moreover, methylcellulose increased the rate of viable spermatozoa with intact acrosome and low lipid disorder, reducing the ROS generation. Preincubation in viscous media increased the penetration rate and the mean number of spermatozoa bound to the zona pellucida (both with 0.5 and 1% MET) and reduced monospermy with 1% MET. On the other hand fertilization in the viscous media reduced penetration rate and increased monospermy. The efficiency of the IVF system was not improved with the use of viscous media. The results show the relevance of increasing viscosity thus making the in vitro media more comparable to physiological conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Melatonin-induced increase of lipid droplets accumulation and in vitro maturation in porcine oocytes is mediated by mitochondrial quiescence.

    Science.gov (United States)

    He, Bin; Yin, Chao; Gong, Yabin; Liu, Jie; Guo, Huiduo; Zhao, Ruqian

    2018-01-01

    Melatonin, the major pineal secretory product, has a significant impact on the female reproductive system. Recently, the beneficial effects of melatonin on mammalian oocyte maturation and embryonic development have drawn increased attention. However, the exact underlying mechanisms remain to be fully elucidated. This study demonstrates that supplementing melatonin to in vitro maturation (IVM) medium enhances IVM rate, lipid droplets (LDs) accumulation as well as triglyceride content in porcine oocytes. Decrease of mitochondrial membrane potential, mitochondrial respiratory chain complex IV activity as well as mitochondrial reactive oxygen species (mROS) content indicated that melatonin induced a decrease of mitochondrial activity. The copy number of mitochondrial DNA (mtDNA) which encodes essential subunits of oxidative phosphorylation (OXPHOS), was not affected by melatonin. However, the expression of mtDNA-encoded genes was significantly down-regulated after melatonin treatment. The DNA methyltransferase DNMT1, which regulates methylation and expression of mtDNA, was increased and translocated into the mitochondria in melatonin-treated oocytes. The inhibitory effect of melatonin on the expression of mtDNA was significantly prevented by simultaneous addition of DNMT1 inhibitor, which suggests that melatonin regulates the transcription of mtDNA through up-regulation of DNMT1 and mtDNA methylation. Increase of triglyceride contents after inhibition of OXPHOS indicated that mitochondrial quiescence is crucial for LDs accumulation in oocytes. Taken together, our results suggest that melatonin-induced reduction in mROS production and increase in IVM, and LDs accumulation in porcine oocytes is mediated by mitochondrial quiescence. © 2017 Wiley Periodicals, Inc.

  3. Development of the isolated perfused porcine skin flap for in vitro studies of percutaneous absorption pharmacokinetics and cutaneous biotransformation

    Energy Technology Data Exchange (ETDEWEB)

    Carver, M.P.

    1988-01-01

    The isolated perfused porcine skin flap (IPPSF) has proven to be a valuable in vitro tool for studying the physiology and biochemistry of skin and for identifying biochemical and histological markers of direct cutaneous toxicity. The present experiments were undertaken for two purposes: (1) to develop a pharmacokinetic model, based on dermal penetration in the IPPSF, which is predictive of percutaneous absorption in vivo, and (2) to examine cutaneous biotransformation of the important agricultural poison parathion (P). Dosing solutions of {sup 14}C-radiolabelled compounds representing 3 chemical classes-organic acid/base (benzoic acid (B), caffeine (C)), organophosphate (OP) pesticides, and steroid hormones, were applied topically in ethanol at 40 {mu}m cm{sup {minus}2}, both in vivo and on the IPPSF. A 3-compartment pharmacokinetic model describing mass transfer from the surface (C{sub 1}), diffusion through epidermis and dermis (C{sub 2}), and transfer into the perfusate (C{sub 3}), was developed based on flux through the IPPSF from 0-8 hr. Model simulations were predictive of percutaneous absorption in vivo for the OP's and steroids. Modification of the basic 3-compartment model to account for fast and slow tissue-release processes (B) and for flux-dependent perfusage flow increases (C), provided excellent in vivo-in vitro correlation over all 7 compounds.

  4. Ectopic pregnancy rates after in vitro fertilization: a look at the donor egg population.

    Science.gov (United States)

    Rosman, Elana R; Keegan, Debbra A; Krey, Lewis; Liu, Mengling; Licciardi, Frederick; Grifo, Jamie A

    2009-11-01

    In an 8-year review of ectopic pregnancy (EP) rates in donor egg recipients and standard patients undergoing in vitro fertilization-embryo transfer (IVF-ET) at a large university-based program, we report an EP rate of 0.6% in donor egg recipients and 0.9% in standard IVF patients, a difference that is not statistically significant. Donor egg recipients were found to have a significantly lower incidence of tubal disease compared with standard IVF patients; however, tubal disease was not found to be an independent risk factor for EP in our practice, perhaps owing to aggressive management of tubal disease.

  5. Drug-Induced Thrombocytopenia following a Transvaginal Oocyte Retrieval for In Vitro Fertilization

    Directory of Open Access Journals (Sweden)

    Ioanna A. Comstock

    2015-01-01

    Full Text Available Drug-induced immune thrombocytopenia has been associated with hundreds of medications and can lead to devastating consequences for the patient. We present a case of a healthy 33-year-old female undergoing in vitro fertilization who developed a severe drug-induced thrombocytopenia, petechiae, and a large hemoperitoneum after receiving Cefazolin antibiotic prophylaxis for a transvaginal oocyte retrieval. The patient was admitted to the intensive care unit for resuscitation with blood products. The presence of drug-dependent platelet antibodies to Cefazolin was confirmed serologically.

  6. Drug-Induced Thrombocytopenia following a Transvaginal Oocyte Retrieval for In Vitro Fertilization.

    Science.gov (United States)

    Comstock, Ioanna A; Longmire, Michelle; Aster, Richard H; Milki, Amin A

    2015-01-01

    Drug-induced immune thrombocytopenia has been associated with hundreds of medications and can lead to devastating consequences for the patient. We present a case of a healthy 33-year-old female undergoing in vitro fertilization who developed a severe drug-induced thrombocytopenia, petechiae, and a large hemoperitoneum after receiving Cefazolin antibiotic prophylaxis for a transvaginal oocyte retrieval. The patient was admitted to the intensive care unit for resuscitation with blood products. The presence of drug-dependent platelet antibodies to Cefazolin was confirmed serologically.

  7. In vitro fertilization complicated by rupture of tubo-ovarian abscess during pregnancy.

    Science.gov (United States)

    Han, Cha; Wang, Chen; Liu, Xiao-Juan; Geng, Nv; Wang, Ying-Mei; Fan, Ai-Ping; Yuan, Bi-Bo; Xue, Feng-Xia

    2015-10-01

    Pelvic abscess during pregnancy is an uncommon complication, but can lead to adverse perinatal outcomes during pregnancy. We present a patient who developed rupture of a tubo-ovarian abscess during pregnancy following in vitro fertilization and embryo transfer. Thirty-eight reported cases are reviewed, and transvaginal oocyte retrieval, genital tract infections, endometrioma, and previous pelvic surgery are considered as risk factors for pelvic abscess during pregnancy. Prolonging gestational duration when an infection situation is allowed is the principle of treatment. Copyright © 2015. Published by Elsevier B.V.

  8. Tubal Pregnancy Associated with Endometrial Carcinoma after In Vitro Fertilization Attempts

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    Yesim Bayoglu Tekin

    2014-01-01

    Full Text Available Endometrial carcinoma is rarely seen during reproductive ages and commonly related to infertility, polycystic ovarian syndrome (PCOS, and obesity. Pregnancy associated endometrial carcinoma is even rarer and this is the second case reported in the literature concerning tubal pregnancy associated endometrial carcinoma. We present a case of a 36-year-old woman with a history of PCOS, infertility, and several attempts of ovulation induction and in vitro fertilization, who was diagnosed with tubal pregnancy and a well differentiated endometrial carcinoma. We also review the literature about pregnancy associated endometrial carcinoma in the first trimester.

  9. Complete in vitro generation of fertile oocytes from mouse primordial germ cells

    Science.gov (United States)

    Morohaku, Kanako; Tanimoto, Ren; Sasaki, Keisuke; Kawahara-Miki, Ryouka; Kono, Tomohiro; Hayashi, Katsuhiko; Hirao, Yuji; Obata, Yayoi

    2016-01-01

    Reconstituting gametogenesis in vitro is a key goal for reproductive biology and regenerative medicine. Successful in vitro reconstitution of primordial germ cells and spermatogenesis has recently had a significant effect in the field. However, recapitulation of oogenesis in vitro remains unachieved. Here we demonstrate the first reconstitution, to our knowledge, of the entire process of mammalian oogenesis in vitro from primordial germ cells, using an estrogen-receptor antagonist that promotes normal follicle formation, which in turn is crucial for supporting oocyte growth. The fundamental events in oogenesis (i.e., meiosis, oocyte growth, and genomic imprinting) were reproduced in the culture system. The most rigorous evidence of the recapitulation of oogenesis was the birth of fertile offspring, with a maximum of seven pups obtained from a cultured gonad. Moreover, cryopreserved gonads yielded functional oocytes and offspring in this culture system. Thus, our in vitro system will enable both innovative approaches for a deeper understanding of oogenesis and a new avenue to create and preserve female germ cells. PMID:27457928

  10. Comparison of normal and abnormal fertilization of in vitro-matured human oocyte according to insemination method.

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    Park, Ju Hee; Jee, Byung Chul; Kim, Seok Hyun

    2016-04-01

    Our purpose was to compare the normal fertilization rate, multi-pronuclei (PN) formation rate, and embryonic development of in vitro-matured oocytes between conventional insemination and intracytoplasmic sperm injection (ICSI). A total of 213 stimulated in vitro fertilization (IVF) cycles were selected, in which at least one immature oocyte was obtained (from 2010 to 2014). Immature oocytes were assigned to germinal vesicle (GV)-stage or metaphase I (MI)-stage oocyte groups. Cycles with obligatory ICSI due to male-factor infertility were excluded. Cycles were divided into two groups according to fertilization method: there were 97 cycles with conventional insemination and 116 cycles with ICSI. After in vitro maturation of 324 GV-stage oocytes and 341 MI-stage oocytes, the fertilization rate, multi-PN formation rate, and embryonic development were compared according to the fertilization method. The normal fertilization rate was similar in the conventional insemination and the ICSI both in GV-derived and MI-derived oocytes. Both fertilization methods resulted in a similar multi-PN formation rate in GV-derived oocytes; however, in MI-derived oocytes, the multi-PN formation rate was zero with ICSI and this was significantly lower than that with conventional insemination (9.6%, P = 0.001). In non-male-factor infertility, ICSI should be considered when MI oocytes are matured. © 2016 Japan Society of Obstetrics and Gynecology.

  11. Intracellular bacteria of porcine proliferative enteropathy: cultivation and maintenance in vitro.

    Science.gov (United States)

    Lawson, G H; McOrist, S; Jasni, S; Mackie, R A

    1993-01-01

    An obligate intracellular bacterium was isolated from the intestines of all 10 cases of porcine proliferative enteropathy from four different pig farms. The organism grew in a rat enterocyte cell line (IEC-18) and was maintained over 20 passages. The growth of the bacteria was assessed by immunostaining of cells exposed to infection. Infection was not associated with morphological cell change, and growth was confined to cells infected at the time of each transfer of infection and the progeny of these cells. The bacterium is a microaerophilic, cell dependent, curved or rod-shaped, gram-negative bacillus that multiplies freely in the enterocyte cytoplasm. Cell cultures containing the intracellular bacteria appear to be free of other microorganisms, including chlamydiae and viruses. Images PMID:8501214

  12. Enhancement of lipid metabolism with L-carnitine during in vitro maturation improves nuclear maturation and cleavage ability of follicular porcine oocytes.

    Science.gov (United States)

    Somfai, Tamás; Kaneda, Masahiro; Akagi, Satoshi; Watanabe, Shinya; Haraguchi, Seiki; Mizutani, Eiji; Dang-Nguyen, Thanh Quang; Geshi, Masaya; Kikuchi, Kazuhiro; Nagai, Takashi

    2011-01-01

    The aim of the present study was to assess the effects of L-carnitine, an enhancer of lipid metabolism and mitochondrial activity, during in vitro maturation (IVM) on nuclear maturation and in vitro fertilisation of porcine follicular oocytes and subsequent embryo development. Mitochondrial functions, intracellular lipid content and reactive oxygen species (ROS) levels in oocytes were also investigated. L-carnitine supplementation in 0.6-5mgmL(-1) concentration during IVM significantly improved (Pcarnitine significantly increased oocyte cleavage (Pmitochondria was significantly higher and the density of lipid droplets was significantly lower (Pcarnitine-treated oocytes compared with the control. Furthermore, the ROS levels in L-carnitine-treated oocytes were significantly lower than those in the control. In conclusion, enhancing mitochondrial functions by L-carnitine improved oocyte maturation and cleavage underlining the importance of lipid metabolism for nuclear and cytoplasmic maturation of porcine oocytes.

  13. Effect of bovine sperm chromatin integrity evaluated using three different methods on in vitro fertility.

    Science.gov (United States)

    Castro, L S; Siqueira, A F P; Hamilton, T R S; Mendes, C M; Visintin, J A; Assumpção, M E O A

    2018-02-01

    In vitro fertility potential of individual bulls is still relatively uncharacterized. Classical sperm analysis does not include the evaluation of all sperm characteristics and thus, some cell compartments could be neglected. In humans, sperm DNA integrity has already proven to have major influence in embryo development and assisted reproduction techniques successfully. In bovine, some studies already correlated chromatin integrity with field fertility. However, none of those have attempted to relate DNA assessment approaches such as chromatin deficiency (CMA3), chromatin stability (SCSA; AO+) and DNA fragmentation (COMET assay) to predict in vitro bull fertility. To this purpose, we selected bulls with high and low in vitro fertility (n = 6/group), based on embryo development rate (blastocyst/cleavage rate). We then performed CMA3, SCSA test and COMET assay to verify if the difference of in vitro fertility may be related to DNA alterations evaluated by these assays. For the three tests performed, our results showed only differences in the percentage of cells with chromatin deficiency (CMA3+; high: 0.19 ± 0.03 vs low: 0.04 ± 0.04; p = 0.03). No difference for chromatin stability and any of COMET assay categories (grade I to grade IV) was observed between high and low in vitro fertility bulls. A positive correlation between AO + cells and grade IV cells was found. Despite the difference between groups in CMA3 analysis, our results suggest that protamine deficiency in bovine spermatozoa may not have a strong biological impact to explain the difference of in vitro fertility between the bulls used in this study. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. The role of oxidative stress in female infertility and in vitro fertilization

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    Joanna Wojsiat

    2017-05-01

    Full Text Available Infertility problem involves many couples of reproductive age. It has been estimated that in Poland 0.7-1.0 million pairs require treatment, while for more than half of them assisted reproduction is the only recommended and effective method. Infertility affects 13 to 15% of the world’s population. A major concern is the age-related decline in female fertility even more that often a decision about pregnancy is taken at later age. Recent studies show that increased production of reactive oxygen species is an important factor in etiopathogenesis of pregnancy and affects female reproduction. It was found that oxidative stress may damage the oocytes and may impair their fertilization capacity. Oxidative stress may also lead to embryo fragmentation and formation of numerous developmental abnormalities, and is regarded to be one of the important reasons of spontaneous and recurrent miscarriage. Moreover, overproduction of reactive oxygen species has a significant impact on the success of in vitro fertilization (IVF.

  15. Comparison of the cutaneous iontophoretic delivery of rasagiline and selegiline across porcine and human skin in vitro.

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    Kalaria, Dhaval R; Patel, Pratik; Patravale, Vandana; Kalia, Yogeshvar N

    2012-11-15

    The objective was to investigate the anodal iontophoresis of the MAO-B inhibitors rasagiline (RAS) and selegiline (SEL) across porcine and human skin in vitro. Passive delivery of RAS and SEL from aqueous solution was minimal; however, increasing current density from 0.1 to 0.3 and 0.5 mA/cm(2) produced a linear increase in steady-state iontophoretic flux (J(ss,RAS)=49.1i(d)+27.9 (r(2)=0.96) and J(ss,SEL)=27.8i(d)+25.8 (r(2)=0.98)). In the absence of background electrolyte, a four-fold change in donor concentration (10, 20 and 40 mM) did not produce a statistically significant increase in cumulative permeation of either drug after iontophoresis at 0.5mA/cm(2) for 7h. Co-iontophoresis of acetaminophen confirmed that electromigration was the dominant transport mechanism for both drugs (∼90%). Total iontophoretic delivery of RAS and SEL across porcine and human skin in vitro was statistically equivalent (RAS: 1512.7 ± 163.7 and 1523.6 ± 195.9 μg/cm(2), respectively, and SEL: 1268.7 ± 231.2 and 1298.3 ± 253.3 μg/cm(2), respectively). Transport efficiencies for RAS and SEL were good (ranged from 6.81 to 8.50 and 2.86 to 3.61%, respectively). Furthermore, the delivery efficiency, i.e., the fraction of the drug in the formulation that was delivered was very high (>56% at 0.5 mA/cm(2)). Cumulative permeation of RAS and SEL from carbopol gels, potential drug reservoirs for iontophoretic systems, was 891.5 ± 148.3 and 626.6 ± 162.4 μg/cm(2), respectively; this was less than from solution and was tentatively attributed to either different partitioning or slower drug diffusion in the gel matrix. The results demonstrated that therapeutic amounts of rasagiline and selegiline could be easily delivered by transdermal iontophoresis with simple gel patches of modest surface area. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Gender and social controversies of in vitro fertilization in Serbia: Discrimination against childless women

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    Kričković-Pele Ksenija

    2014-01-01

    Full Text Available This paper analyses gender and social controversies of assisted reproductive technologies and the discrimination of childless women in Serbia. Primary goals of this paper are critical analysis of new reproductive technologies phenomenon, discrimination against women without children and critical analysis of the legal framework regulating biomedical assisted reproduction in Serbia from gender studies and feminist methodology perspectives, as well as presentation of the research results on discrimination of childless women. For the purpose of this research the survey and the content analysis have been used. A survey was conducted of 50 female participants in the in vitro fertilization program at the Department for Gynecology and Obstetrics in Novi Sad. The results indicate that the regulations on biomedical assisted reproduction and the criteria for inclusion in the in vitro fertilization program are discriminatory and that women involved in the program feel discriminated against, usually at work and in their own surroundings. The conclusion is that it is necessary to change the regulations governing this area, further work on the elimination of discrimination against childless women and destigmatisation of women and couples that cannot or do not want to have children.

  17. PRELIMINARY CLINICAL OBSERVATIONS ON THE GINGIVAL-PERIODONTAL MANIFESTATIONS IN IN VITRO FERTILIZED FEMALE PATIENTS

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    Ana GIURA

    2012-06-01

    Full Text Available Scope of the study – determination of the types and intensity of the gingival-periodontal manifestations in in vitro fertilized pregnant women, and establishment of the role played by the hormonal treatment in the initiation and maintenance of such manifestations. Materials and method. The study was performed on patients fertilized in vitro between the years 2010-2012, in the Section of Assisted Human Reproduction of the “Panait Sârbu” Clinical Hospital of Obstretics and Ginecology – Bucuresti. All patients received the same amount and concentration of progesteronic compounds for maintaining their state of pregnancy, being examined in the first gestation weeks, immediately after confirmation of pregnancy through the β-hCG blood test. Results anddiscussion: In the last decades, the effects of estrogen and progesterone have drawn special attention from the part of the researchers. During pregnancy, the gingiva represents the maintissular target for the action of steroid hormones. Pregnancy-induced gingivitis is directly correlated with the presence of the microbial flora, known as the determining factor of the disease, being still exacerbated by the action of the sexual hormones, especially during the second and third quarter of pregnancy. Conclusions: The influence of gestational hormones upon the immune system of the organism may further contribute to the initiation and worsening of pregnancy-induced gingivitis, by reducing the immune response vs. the bacterial plaque.

  18. Acute intermittent porphyria exacerbation following in vitro fertilization treatment.

    Science.gov (United States)

    New, Geok Huey; Hsu, Pei-Yang; Wu, Meng-Hsing

    2016-04-01

    Assisted reproductive technology is commonly used for women with infertility. We report a case of acute intermittent porphyria associated with in vitro fertilization treatment. A 35-year-old woman with tubal factor infertility presented to our clinic with persistent low abdominal pain and hyponatremia after transvaginal oocyte retrieval. During admission, she experienced a generalized tonic-clonic seizure attacked following by dark brown color urine. Urinary tests showed elevated porphobilinogen, 5-aminolevulinic acid, uroporphyrin, and coproporphyrin, confirming the diagnosis of acute intermittent porphyria. The patient's condition continued to improve after hemin treatment and rehabilitation. Newly onset acute intermittent porphyria during the course of controlled ovarian hyperstimulation for in vitro fertilization is a rare but possible complication. Acute intermittent porphyria should be taken into consideration for persisted unexplained abdominal pain and seriously alerted if accompanied with neurological symptoms. Special tests for acute intermittent porphyria should be taken into consideration for the differential diagnosis of lower abdominal pain after oocyte retrieval. Copyright © 2016. Published by Elsevier B.V.

  19. Porcine circovirus type 2 increases interleukin-1beta and interleukin-10 production via the MyD88-NF-kappa B signaling pathway in porcine alveolar macrophagesin vitro.

    Science.gov (United States)

    Han, Junyuan; Zhang, Shuxia; Zhang, Yaqun; Chen, Mengmeng; Lv, Yingjun

    2017-06-30

    Porcine alveolar macrophages (PAMs) represent the first line of defense in the porcine lung after infection with porcine circovirus type 2 (PCV2) via the respiratory tract. However, PCV2 infection impairs the microbicidal capability of PAMs and alters cytokine production and/or secretion. At present, the reason for the imbalance of cytokines has not been fully elucidated, and the regulatory mechanisms involved are unclear. In this study, we investigated the expression levels and regulation of interleukin-1beta (IL-1β) and IL-10 in PAMs following incubation with PCV2 in vitro. Levels of IL-1β and IL-10 increased in PAM supernatants, and the distribution of nuclear factor kappa B (NF-κB) p65staining in nucleus, expression of MyD88 and p-IκB in cytoplasm, and DNA-binding activity of NF-κB increased after incubation with PCV2, while p65 expression in PAM cytoplasm decreased. However, when PAMs were co-incubated with PCV2 and small interfering RNA targeting MyD88, those effects were reversed. Additionally, mRNA expression levels of Toll-like receptors (TLR)-2, -3, -4, -7, -8, and -9 increased when PAMs were incubated with PCV2. These results show that PCV2 induces increased IL-1β and IL-10 production in PAMs, and these changes in expression are related to the TLR-MyD88-NF-κB signaling pathway.

  20. In vitro culture and in vitro fertilization techniques for prairie voles (Microtus ochrogaster).

    Science.gov (United States)

    Horie, Kengo; Hidema, Shizu; Hirayama, Takashi; Nishimori, Katsuhiko

    2015-08-07

    Prairie vole (Microtus ochrogaster) is a highly social animal and is a commonly used animal model for neuropsychopharmacological and psychiatric studies. To date, only a few reports on the development of transgenic prairie voles which was primarily due to the suboptimal development of assisted reproductive technology (ART) in prairie voles. Limitations in ART further hinder the development of genetically modified prairie voles such as the application of conventional gene targeting technologies using embryonic stem (ES) or induced pluripotent stem (iPS) cells to generate chimeric prairie voles. Moreover, recent advancement in genome-editing tools such as transcription activator-like effector nuclease (TALEN) and clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas technology provide an unprecedented opportunity to create gene targeting animal model and the development of ART in prairie voles is necessary for future development of novel transgenic prairie vole model. We have established efficient method for in vitro embryo culture and sperm cryopreservation with high fertilization rate. In G-1 PLUS and G-2 PLUS sequential culture condition, 81.0% (# of Blastocysts/total n) of one-cell embryos developed to blastocysts. In contrary, no embryos were developed to blastocyst stage in KSOM medium (0/total # of embryos in culture). In vitro fertilization rate using fresh and frozen-thawed sperm was 32.6% and 29.3%, respectively. This is the first report of IVF using cryopreserved prairie vole sperm. We employed mouse IVF methods in prairie voles and optimize culture conditions using human G-1/G-2 PLUS sequential culture method that resulted in high embryonic development rate. The development in vole reproductive technology will facilitate the generation of transgenic voles in the future. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. First llama (Lama glama) pregnancy obtained after in vitro fertilization and in vitro culture of gametes from live animals.

    Science.gov (United States)

    Trasorras, V; Baca Castex, C; Alonso, A; Giuliano, S; Santa Cruz, R; Arraztoa, C; Chaves, G; Rodríguez, D; Neild, D; Miragaya, M

    2014-07-01

    The aim of this study was to evaluate the developmental competence and pregnancy rate of llama hatched blastocysts produced in vitro using gametes from live animals and two different culture conditions. Fifteen adult females were superstimulated with 1500 IU of eCG, eleven (73%) responded to the treatment and were used as oocyte donors. Follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. Sixty-six COCs were recovered from 77 aspirated follicles (86% recovery) and were randomly placed in Fertil-TALP microdroplets with the sperm suspension (20 × 10(6)live spermatozoa/ml). After 24 h, they were placed in SOFaa medium supplemented with FCS and randomly assigned to one of two culture conditions. Culture condition 1 (CC1) consisted of 6 days of culture (n=28) and culture condition 2 (CC2) consisted of renewing the culture medium every 48 h (n=35). In CC1, the blastocyst rate was 36% (10/28) and the hatched blastocyst rate was 28% (8/28) whereas in CC2, the blastocyst rate was 34% (12/35) and the hatched blastocyst rate was 20% (7/35) (p>0.05). No pregnancies were obtained after embryo transfer (ET) of CC1 blastocysts (0/8) while one pregnancy was obtained (1/7) after transferring a hatched blastocyst from CC2. Forty-two days after the ET, the pregnancy was lost. This study represents the first report of a pregnancy in the llama after intrauterine transfer of embryos produced by in vitro fertilization using gametes from live animals. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. The association between fatty acid index and in vitro fertilization outcomes.

    Science.gov (United States)

    Eskew, Ashley M; Wormer, Kelly C; Matthews, Michelle L; Norton, H James; Papadakis, Margaret A; Hurst, Bradley S

    2017-12-01

    Fatty acids have been shown to play an important role in oocyte competence and early implantation of the embryo. Our hypothesis-generating study sought to determine if individual fatty acids expressed as a percentage of total erythrocyte fatty acids are associated with embryo quality and other in vitro fertilization (IVF) outcomes. This was a prospective cohort study at an academic fertility center. Sixty women undergoing their first IVF cycle were recruited. Serum measurements of 22 fatty acids were obtained. We calculated each fatty acid as a percentage of total fatty acids, defined as the index for that individual fatty acid. Omega-3 index had no correlation with IVF outcomes. A negative correlation was found between the trans fatty acid index, elaidic acid (EA), and IVF outcomes, including fertilization rate (r = - 0.261, p = 0.04), blastocyst conversion rate (r = - 0.41, p = 0.001), and number of usable blastocysts and embryos (r = - 0.411, p = 0.001). There was no correlation between EA index and number of oocytes retrieved, embryo grade, or clinical pregnancy. No consistent correlations were observed with the additional fatty acids analyzed. No correlation was observed between omega-3 index and IVF outcomes. Elevated erythrocyte EA index, the major trans fatty acid commonly consumed in hydrogenated oils, margarine, and fried foods, was negatively correlated with number of usable blastocysts and embryos, blastocyst conversion, and fertilization rate. Our findings suggest preliminary evidence that trans fat may be negatively associated with IVF outcomes.

  3. Protective effect of crocetin on bovine spermatozoa against oxidative stress during in vitro fertilization.

    Science.gov (United States)

    Sapanidou, V; Taitzoglou, I; Tsakmakidis, I; Kourtzelis, I; Fletouris, D; Theodoridis, A; Lavrentiadou, S; Tsantarliotou, M

    2016-11-01

    Oxidative stress is one of the major factors that contribute to poor semen quality and low rates of in vitro fertilization. Crocetin, a main constituent of saffron (Crocus sativus L.) possesses potent antioxidant activity, by scavenging reactive oxygen species (ROS) and/or enhancing the activity of intracellular antioxidant enzymes. The aim of this study was to investigate, for the first time, the effect of crocetin on the quality characteristics of bull spermatozoa and fertilization rate. For this reason, frozen/thawed bovine spermatozoa were incubated with crocetin (1, 2.5, and 5 μm), for 120 or 240 min, in the presence of a negative control, and evaluated in terms of motility, viability, acrosomal status, DNA fragmentation index, intracellular ROS, and lipid peroxidation. In order to evaluate the impact of crocetin on cleavage and blastocyst rate, the compound was added in the IVF medium at the previously identified optimal concentration (2.5 μm). The results indicate that incubation of spermatozoa with 2.5 μm of crocetin resulted in a statistically significant lower production of superoxide anion and hydrogen peroxide, lower lipid peroxidation, and in better maintenance of motility parameters, viability, and acrosomal integrity, with a very small number of cells with DNA fragmentation, compared to the other groups (p fertilization medium also resulted in a significant increase in acrosome-reacted spermatozoa and blastocyst production, compared to the control group (p fertilizing ability, directly and/or indirectly, by regulating ROS concentration and lipid peroxidation. © 2016 American Society of Andrology and European Academy of Andrology.

  4. Effective inhibition of porcine epidemic diarrhea virus by RNA interference in vitro.

    Science.gov (United States)

    Shen, Haiyan; Zhang, Chunhong; Guo, Pengju; Liu, Zhicheng; Zhang, Jianfeng

    2015-10-01

    Porcine epidemic diarrhea virus (PEDV) is a member of the coronaviridae family, which can cause acute and highly contagious enteric disease of swine characterized by severe entero-pathogenic diarrhea in piglets. Currently, the vaccines of PEDV are only partially effective and there is no specific drug available for treatment of PEDV infection. To exploit the possibility of using RNA interference (RNAi) as a strategy against PEDV infection, five shRNA-expressing plasmids targeting the N, M, and S genes of PEDV were constructed and transfected into Vero cells. The cytopathic effect and MTS assays demonstrated that two shRNAs (pSilencer4.1-M1 and pSilencer4.1-N) were capable of protecting cells against PEDV invasion with very high specificity and efficiency. The two shRNA expression plasmids were also able to inhibit the PEDV replication significantly, as shown by detection of virus titers (TCID50/mL). A real-time quantitative RT-PCR further confirmed that the amounts of viral RNAs in cell cultures pre-transfected with these two plasmids were reduced by 95.0 %. Our results suggest that RNAi might be a promising new strategy against PEDV infection.

  5. Aflatoxins of type B and G affect porcine dendritic cell maturation in vitro.

    Science.gov (United States)

    Mehrzad, Jalil; Devriendt, Bert; Baert, Kim; Cox, Eric

    2015-01-01

    The toxic effects of highly carcinogenic mycotoxins, especially aflatoxins (AF), on key antigen-presenting cells, such as dendritic cells (DC), are largely unknown. To elucidate the effect of AF on DC function, porcine monocyte-derived DC (MoDC) were treated with a mixture of several AF (i.e., AFB1, AFB2, AFG1, and AFG2) and the phagocytic capacity, the membrane expression level of several DC activation markers, the T-cell proliferation-inducing capacity, and the cytokine secretion pattern were assessed. As compared to untreated MoDC, AF significantly up-regulated the expression of the co-stimulatory molecules CD25 and CD80/86. However, the phagocytic activity of MoDC was not affected by AF treatment. While the cytokine secretion pattern of AF-treated MoDC was similar to control MoDC, the T-cell proliferation-inducing capacity of MoDC was increased upon aflatoxin treatment. The results indicate that a mixture of naturally occurring AF enhances the antigen-presenting capacity of DC, which could explain the observed immunotoxicity of AF by breaking down tolerance and further emphasizes the need to reduce the admissible level of AF in agricultural commodities.

  6. Interaction of porcine circovirus type 2 replication with intracellular redox status in vitro.

    Science.gov (United States)

    Chen, Xingxiang; Ren, Fei; Hesketh, John; Shi, Xiuli; Li, Junxian; Gan, Fang; Hu, Zhihua; Huang, Kehe

    2013-01-01

    Redox status influences replication of some viruses but its effect on porcine circovirus type 2 (PCV2), the primary causative agent of the emerging swine disease post-weaning multisystemic wasting syndrome is not known. The interaction of PCV2 replication with intracellular redox status in PK15 cells was examined in this study. Intracellular glutathione (GSH) was measured spectrophotometrically by reaction with 5, 5'-dithiobis (2-nitrobenzoic acid). Total superoxide dismutase activity (SOD) was assayed by inhibition of oxyamine oxidation by the xanthine oxidase system. Malondialdehyde (MDA) was assayed spectrophotometrically using the thiobarbituric acid reaction. Both quantification of PCV2 DNA by real-time polymerase chain reaction and indirect immunofluorescence of PCV2-infected cells were used to evaluate the replication of PCV2. Both GSH and SOD decreased significantly at 48 hours after PCV2 infection, whereas MDA concentration increased significantly after 48 hour post-infection. Furthermore, PCV2 replication in PK15 cells was significantly impaired after the elevation of intracellular GSH through treatment with the antioxidant N-acetyl-l-cysteine (NAC), a precursor in GSH synthesis. In contrast, PCV2 replication in PK15 cells was enhanced after reduction of GSH levels through H2O2-mediated oxidation. In addition, NAC treatment blocked the increase of virus replication induced by H2O2. This study suggests that PCV2 infection induces oxidative stress and that intracellular redox status influences PCV2 replication in PK15 cells.

  7. Determination of in vitro usnic acid delivery into porcine skin using a HPLC method.

    Science.gov (United States)

    Serafini, Mairim Russo; Detoni, Cassia Britto; Guterres, Sílvia Stanisçuaski; da Silva, Gabriel Francisco; de Souza Araújo, Adriano Antunes

    2015-01-01

    Usnic acid, a lichen metabolite, has been proposed as a potential topical treatment for microbial skin lesions, burn wounds as well as a sunscreen. An isocratic HPLC method was validated according to FDA's Guidance for Industry: Bioanalytical Method Validation to determine skin penetration and permeation of usnic acid. The penetration and permeation of usnic acid was evaluated using Franz cells and porcine skin. The method was valid according to selectivity, linearity, precision, accuracy and stability. Usnic acid was quantified in the skin surface (6.13 µg cm(2)), stratum corneum (34.4 µg cm(2)), viable epidermis (5.6 µg cm(2)), dermis (28.2 µg cm(2)) and receptor compartment (3.2 µg cm(2)). These results help us to understand the penetration profile of usnic acid and plan topical therapeutic approaches as well as new topical delivery systems to modulate this penetration profile. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  8. The role of in vitro fertilization and intracytoplasmic sperm injection in couples with unexplained infertility after failed intrauterine insemination.

    Science.gov (United States)

    Ruiz, A; Remohí, J; Minguez, Y; Guanes, P P; Simón, C; Pellicer, A

    1997-07-01

    To determine an optimal insemination technique in patients undergoing IVF after failed IUI and the role of intracytoplasmic sperm injection (ICSI) in such cases. Prospective, randomized study in couples with unexplained infertility (n = 63) and mild endometriosis (n = 7) undergoing IVF after four IUI cycles. Sibling oocytes were randomized into standard IVF or ICSI insemination according to the order of retrieval. In vitro fertilization program at the Instituto Valenciano de Infertilidad, Valencia, Italy. Seventy couples with unexplained infertility undergoing IVF after failing to conceive with controlled ovarian stimulation and IUI. In vitro fertilization and ICSI. Fertilization, cleavage, and embryo quality were compared in IVF- and ICSI-inseminated oocytes. There was no significant difference in fertilization rates between ICSI (60.4%) and conventional IVF (54.0%). Similarly, there was no difference in embryo quality between both groups. There was no total fertilization failure in ICSI-inseminated oocytes, whereas 8 (11.4%) of 70 cases showed absence of fertilization when conventional IVF was used. Couples with unexplained infertility and mild endometriosis failing to conceive with IUI and undergoing IVF have an 11.4% chance of fertilization failure that can be overcome easily by using ICSI in at least some oocytes. ICSI, however, is not superior to IVF as an insemination technique in most cases. These data should be used in counseling patients.

  9. Changes in chondrocyte gene expression following in vitro impaction of porcine articular cartilage in an impact injury model.

    Science.gov (United States)

    Ashwell, Melissa S; Gonda, Michael G; Gray, Kent; Maltecca, Christian; O'Nan, Audrey T; Cassady, Joseph P; Mente, Peter L

    2013-03-01

    Our objective was to monitor chondrocyte gene expression at 0, 3, 7, and 14 days following in vitro impaction to the articular surface of porcine patellae. Patellar facets were either axially impacted with a cylindrical impactor (25 mm/s loading rate) to a load level of 2,000 N or not impacted to serve as controls. After being placed in organ culture for 0, 3, 7, or 14 days, total RNA was isolated from full thickness cartilage slices and gene expression measured for 17 genes by quantitative real-time RT-PCR. Targeted genes included those encoding proteins involved with biological stress, inflammation, or anabolism and catabolism of cartilage extracellular matrix. Some gene expression changes were detected on the day of impaction, but most significant changes occurred at 14 days in culture. At 14 days in culture, 10 of the 17 genes were differentially expressed with col1a1 most significantly up-regulated in the impacted samples, suggesting impacted chondrocytes may have reverted to a fibroblast-like phenotype. Copyright © 2012 Orthopaedic Research Society.

  10. Assessing the Impact of Mechanical Damage on Full-Thickness Porcine and Human Skin Using an In Vitro Approach

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    Hinda Dabboue

    2015-01-01

    Full Text Available For most xenobiotics, the rates of percutaneous absorption are limited by diffusion through the horny layer of skin. However, percutaneous absorption of chemicals may seriously increase when the skin is damaged. The aim of this work was to develop an in vitro representative model of mechanically damaged skins. The epidermal barrier was examined following exposure to a razor, a rotating brush, and a microneedle system in comparison to tape-stripping which acted as a reference. Excised full-thickness skins were mounted on a diffusion chamber in order to evaluate the effect of injuries and to mimic physiological conditions. The transepidermal water loss (TEWL was greatly increased when the barrier function was compromised. Measurements were made for all the damaged biopsies and observed histologically by microscopy. On human and porcine skins, the tape-stripping application (0 to 40 times showed a proportional increase in TEWL which highlights the destruction of the stratum corneum. Similar results were obtained for all cosmetic instruments. This is reflected in our study by the nonsignificant difference of the mean TEWL scores between 30 strips and mechanical damage. For a specific appreciation, damaged skins were then selected to qualitatively evaluate the absorption of a chlorogenic acid solution using fluorescence microscopy.

  11. Assessing the Impact of Mechanical Damage on Full-Thickness Porcine and Human Skin Using an In Vitro Approach.

    Science.gov (United States)

    Dabboue, Hinda; Builles, Nicolas; Frouin, Éric; Scott, Dan; Ramos, Jeanne; Marti-Mestres, Gilberte

    2015-01-01

    For most xenobiotics, the rates of percutaneous absorption are limited by diffusion through the horny layer of skin. However, percutaneous absorption of chemicals may seriously increase when the skin is damaged. The aim of this work was to develop an in vitro representative model of mechanically damaged skins. The epidermal barrier was examined following exposure to a razor, a rotating brush, and a microneedle system in comparison to tape-stripping which acted as a reference. Excised full-thickness skins were mounted on a diffusion chamber in order to evaluate the effect of injuries and to mimic physiological conditions. The transepidermal water loss (TEWL) was greatly increased when the barrier function was compromised. Measurements were made for all the damaged biopsies and observed histologically by microscopy. On human and porcine skins, the tape-stripping application (0 to 40 times) showed a proportional increase in TEWL which highlights the destruction of the stratum corneum. Similar results were obtained for all cosmetic instruments. This is reflected in our study by the nonsignificant difference of the mean TEWL scores between 30 strips and mechanical damage. For a specific appreciation, damaged skins were then selected to qualitatively evaluate the absorption of a chlorogenic acid solution using fluorescence microscopy.

  12. In Vitro Coinfection and Replication of Classical Swine Fever Virus and Porcine Circovirus Type 2 in PK15 Cells.

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    Niu Zhou

    Full Text Available Increasing clinical lines of evidence have shown the coinfection/superinfection of porcine circovirus type 2 (PCV2 and classical swine fever virus (CSFV. Here, we investigated whether PCV2 and CSFV could infect the same cell productively by constructing an in vitro coinfection model. Our results indicated that PCV2-free PK15 cells but not ST cells were more sensitive to PCV2, and the PK15 cell line could stably harbor replicating CSFV (PK15-CSFV cells with a high infection rate. Confocal and super-resolution microscopic analysis showed that PCV2 and CSFV colocalized in the same PK15-CSFV cell, and the CSFV E2 protein translocated from the cytoplasm to the nucleus in PK15-CSFV cells infected with PCV2. Moreover, PCV2-CSFV dual-positive cells increased gradually in PK15-CSFV cells in a PCV2 dose-dependent manner. In PK15-CSFV cells, PCV2 replicated well, and the production of PCV2 progeny was not influenced by CSFV infection. However, CSFV reproduction decreased in a PCV2 dose-dependent manner. In addition, cellular apoptosis was not strengthened in PK15-CSFV cells infected with PCV2 in comparison with PCV2-infected PK15 cells. Moreover, using this coinfection model we further demonstrated PCV2-induced apoptosis might contribute to the impairment of CSFV HCLV strain replication in coinfected cells. Taken together, our results demonstrate for the first time the coinfection/superinfection of PCV2 and CSFV within the same cell, providing an in vitro model to facilitate further investigation of the underlying mechanism of CSFV and PCV2 coinfection.

  13. The in vitro effect of eptifibatide, a glycoprotein IIb/IIIa antagonist, on various responses of porcine blood platelets.

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    Ciborowski, Michał; Tomasiak, Marian

    2009-01-01

    The current study systematically evaluates the in vitro effect of eptifibatide, a GPIIb/IIIa blocker, on various responses of porcine platelets evoked by principal physiological stimulators. Eptifibatide at concentrations up to 40 mg/mL did not affect the calcium signal produced by thrombin, partly reduced the procoagulant response evoked by collagen, and strongly inhibited (IC50 approximately 11 mg/mL) adhesion of these cells to fibrinogen coated surfaces. Eptifibatide in a concentration-dependent manner reduced ADP, collagen, and thrombin-induced platelet aggregation (IC50 = 16-27 mg/mL), dense granule secretion (IC50 = 22-31 mg/mL) and lysosome secretion (IC50 = 25-50 mg/mL). Substantial (up to 30-40%) collagen or thrombin-evoked platelet aggregation still occurred at high (52 mg/mL) eptifibatide concentrations. Direct comparison of the susceptibility of platelet aggregation and dense granule secretion to the inhibitory action of eptifibatide indicates that aggregation is appreciably more sensitive than secretion. Eptifibatide (8 mg/mL) added together with a low (70 ng/mL) concentration of bivalirudin (a direct thrombin inhibitor) effectively (approximately 90%) reduced platelet aggregation induced by thrombin (0.2 U/mL). Based on these results, eptifibatide is not expected to reduce efficiently thrombus formation initiated by rapid local production of large amounts of thrombin. One practical consequence of our in vitro studies is the suggestion that the anti-thrombotic efficacy of eptifibatide, especially in preventing acute thrombotic events, may be largely improved by its combination with direct thrombin inhibitors.

  14. Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos.

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    Pereira, A F; Melo, L M; Freitas, V J F; Salamone, D F

    2015-08-01

    In vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (γH2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring γH2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of γH2AX foci (606.1 ± 103.2) and greater area of γH2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development.

  15. Peroxidized mineral oil increases the oxidant status of culture media and inhibits in vitro porcine embryo development.

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    Martinez, C A; Nohalez, A; Ceron, J J; Rubio, C P; Roca, J; Cuello, C; Rodriguez-Martinez, H; Martinez, E A; Gil, M A

    2017-11-01

    The use of oils with undetected alterations is a long-recognized problem for in vitro embryo production systems. Since peroxides in oils have been associated with reduced embryo production outcomes, our goals were (1) to evaluate the effects of a batch of mineral oil (MO) that was suspected to be altered on the in vitro production of pig embryos and (2) to determine oil peroxide values throughout culture and the transfer of oxidant agents from oil to culture media. Sunflower oil, which has a completely different chemical composition than MO but a higher oxidative status, and unaltered MO were used as controls. Oocyte maturation, fertilization and embryo development were affected differently depending on the oil overlay used. While the suspected MO was not able to sustain in vitro maturation and fertilization, the oocytes incubated in the presence of sunflower oil were matured and fertilized similarly to those of the unaltered MO group. Moreover, the cleavage rate of presumed zygotes cultured under the suspected MO was severely reduced compared with those cultured under the other oils, and none of the cleaved embryos developed to the blastocyst stage. Although the cleavage rates in the sunflower oil and unaltered MO groups were similar, embryos cultured under sunflower oil also failed to develop to the blastocyst stage. Our results revealed that the suspected MO and sunflower oil had similar levels of peroxides and that these levels were much higher than those of the unaltered MO. The total oxidant status was higher in media incubated under peroxidized oils than in fresh media or media incubated without an oil overlay or under unaltered MO, indicating that oxidant agents were transferred to the incubation media. However, unlike the sunflower oil group, the culture media incubated under the suspected MO had high levels of total oxidant status and low levels of hydrogen peroxide and reactive oxygen species, suggesting the presence of other unknown oxidant agents in

  16. Uterine adenomyosis and infertility, review of reproductive outcome after in vitro fertilization and surgery.

    Science.gov (United States)

    Dueholm, Margit

    2017-06-01

    This review includes an analysis of the clinical studies evaluating reproductive outcome and adenomyosis, and a review of studies on reproductive outcome and surgical treatment options for adenomyosis. Strict diagnostic criteria and classification of disease are needed for an image diagnosis of adenomyosis. Studies of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) populations and women with surgically treated deep endometriosis have suggested that adenomyosis has a negative impact on reproductive outcome, although there are substantial variations between studies. Few data are available on the relation between the extent of disease and impact on reproductive outcome, but a correlation appears to exist. Case series seem to confirm a positive effect of gonadotropin-releasing hormone analog treatment and surgery on reproductive outcome, but there are no controlled trials. Evidence is impaired by the poor quality of many studies, a lack of strict image diagnosis, and the absence of a classification of the extent of disease. Selection of the optimal evidence-based treatment options for adenomyosis in the fertility clinic is difficult because of a lack of evidence regarding the relation between fertility and the degree and composition of adenomyosis. Adenomyosis may reduce implantation so severely that surgical or other treatment options should be recommended, but the benefit of these treatment options needs to be verified. Referral of women with adenomyosis and recurrent miscarriage and repeated failure of assisted reproductive technology to centers with a special interest in adenomyosis research and treatment may be critical. © 2017 Nordic Federation of Societies of Obstetrics and Gynecology.

  17. Clinical Outcomes of In Vitro Fertilization among Chinese Infertile Couples Treated for Syphilis Infection.

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    Jing Wang

    Full Text Available To compare the clinical outcomes of infertile patients with and without syphilis after in vitro fertilization and embryo transfer (IVF-ET, in this case-control study, 320 infertile couples were enrolled and divided into syphilis (n = 160 and control groups (n = 160. The primary IVF outcomes were the clinical pregnancy rate and the birth of a healthy baby. All syphilis patients received the standard anti-syphilis treatment before undergoing IVF/ICSI. Our results showed that the endometrial thickness of the syphilis group was greater than that of the control group following hCG injection (16.9±5.4 vs. 13.0±4.7 mm, P<0.001. The numbers of normally fertilized eggs and normally cleaved fertilized eggs and the implantation rate were 6.8±4.8, 6.3±4.7 and 24.2%, respectively, for the syphilis group and 8.3±4.6, 8.1±4.6 and 34.4%, respectively, for the control group, and these values were significantly different between the groups. The clinical pregnancy rate was lower in the syphilis group compared with that in the control group (43.8% vs. 55.6%, P = 0.03. Lower offspring birth weight was observed in the infected male group compared with those in the infected female (2.7±0.4 vs. 3.0±0.4 kg, P = 0.01 and infected couple groups (2.7±0.4 vs. 3.1±0.5 kg, P = 0.007. Therefore, syphilis infection reduces the clinical pregnancy rate after IVF/ICSI.

  18. Influence of strict sperm morphology on the results of classic in vitro fertilization

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    Milton Ghirelli-Filho

    2012-08-01

    Full Text Available OBJECTIVE: To determine the influence of the concentration of oval spermatozoa according to the strict morphology criterion in men with normal sperm concentration following the World Health Organization criteria on the results of classic IVF. MATERIALS AND METHODS: Based on review of patient charts, this study included infertile couples presenting with female causes for infertility or unexplained infertility, in whom men presented with normal spermogram values for sperm concentration, sperm motility, volume of ejaculate and total sperm count after semen processing greater than 20 million. Based on the value obtained in strict sperm morphology, patients were divided into three groups: in Group A, patients with values between 0% and 4%; in group B, between 5% and 14%, and in group C, patients with sperm morphology greater than 14%. The outcomes analyzed were oocyte fertilization rate, biochemical pregnancy rate, clinical pregnancy rate and rate of liveborns. RESULTS: A total of 244 cases met the inclusion criteria, 27 of them in group A, 165 in group B, and 52 in group C. The mean fertilization rate and the rate of liveborns were, respectively: 71.9% and 33.3% in group A; 80.9% and 24.2% in group B, and 78.8% and 28.8% in group C. There was no statistical difference among the groups in any of the outcomes analyzed. CONCLUSION: The values of strict sperm morphology, as proposed by Kruger and adopted by the World Health Organization, had no influence on the results of classic in vitro fertilization in the studied sample.

  19. The creation of "monsters": the discourse of opposition to in vitro fertilization in Poland.

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    Radkowska-Walkowicz, Magdalena

    2012-12-01

    In Poland, there is a campaign to criminalise in vitro fertilization, led by the Catholic church. This article explores how this campaign makes "monsters" of IVF children in its discourse, that is, embodiments of "the other" in the sense of Frankenstein's monster. Basing the analysis primarily on Catholic mass media publications, the article investigates the discursive strategies employed to oppose IVF, most notably by the Catholic clergy and activists and journalists associated with the Church. They attribute "monstrosity" to the children in the following ways: physical (possible bodily deformity), psychological (survivor syndrome, identity crisis), social (loneliness, uncertain place in family relations), and ethical (a life burdened with the deaths of many embryos). Although the world of families with IVF does not provide examples of children who could be considered monsters in any of these terms, these arguments have become the primary reasons given for banning IVF. Copyright © 2012 Reproductive Health Matters. Published by Elsevier Ltd. All rights reserved.

  20. International regulatory landscape and integration of corrective genome editing into in vitro fertilization.

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    Araki, Motoko; Ishii, Tetsuya

    2014-11-24

    Genome editing technology, including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas, has enabled far more efficient genetic engineering even in non-human primates. This biotechnology is more likely to develop into medicine for preventing a genetic disease if corrective genome editing is integrated into assisted reproductive technology, represented by in vitro fertilization. Although rapid advances in genome editing are expected to make germline gene correction feasible in a clinical setting, there are many issues that still need to be addressed before this could occur. We herein examine current status of genome editing in mammalian embryonic stem cells and zygotes and discuss potential issues in the international regulatory landscape regarding human germline gene modification. Moreover, we address some ethical and social issues that would be raised when each country considers whether genome editing-mediated germline gene correction for preventive medicine should be permitted.

  1. Prediction models in in vitro fertilization; where are we? A mini review

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    Laura van Loendersloot

    2014-05-01

    Full Text Available Since the introduction of in vitro fertilization (IVF in 1978, over five million babies have been born worldwide using IVF. Contrary to the perception of many, IVF does not guarantee success. Almost 50% of couples that start IVF will remain childless, even if they undergo multiple IVF cycles. The decision to start or pursue with IVF is challenging due to the high cost, the burden of the treatment, and the uncertain outcome. In optimal counseling on chances of a pregnancy with IVF, prediction models may play a role, since doctors are not able to correctly predict pregnancy chances. There are three phases of prediction model development: model derivation, model validation, and impact analysis. This review provides an overview on predictive factors in IVF, the available prediction models in IVF and provides key principles that can be used to critically appraise the literature on prediction models in IVF. We will address these points by the three phases of model development.

  2. Raman spectrum: A potential biomarker for embryo assessment during in vitro fertilization.

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    Ding, Jiayi; Xu, Tian; Tan, Xiaofang; Jin, Hua; Shao, Jun; Li, Haibo

    2017-05-01

    The aim of the study was to investigate whether Raman spectrum is consistent with the morphological scoring of the embryo of day 3 during in vitro fertilization (IVF). The spent culture media of embryo of day 3 from 10 patients were collected and analyzed. The samples were analyzed using Raman spectroscopy and graded according to the standard embryo scoring system simultaneously. Data showed that the Raman spectra obtained from the droplet of media were useful, as they can act as the characteristic signature for protein and amino acids. The Raman biospectroscopy-based metabonomics profiling of spent media was consistent with the result of conventional morphological evaluation. In conclusion, this technology offers great potential for the development of tools allowing rapid non-invasive assessment of the quality of embryo of day 3 during IVF.

  3. Release of sICAM-1 in oocytes and in vitro fertilized human embryos.

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    Monica Borgatti

    Full Text Available During the last years, several studies have reported the significant relationship between the production of soluble HLA-G molecules (sHLA-G by 48-72 hours early embryos and an increased implantation rate in IVF protocols. As consequence, the detection of HLA-G modulation was suggested as a marker to identify the best embryos to be transferred. On the opposite, no suitable markers are available for the oocyte selection.The major finding of the present paper is that the release of ICAM-1 might be predictive of oocyte maturation. The results obtained are confirmed using three independent methodologies, such as ELISA, Bio-Plex assay and Western blotting. The sICAM-1 release is very high in immature oocytes, decrease in mature oocytes and become even lower in in vitro fertilized embryos. No significant differences were observed in the levels of sICAM-1 release between immature oocytes with different morphological characteristics. On the contrary, when the mature oocytes were subdivided accordingly to morphological criteria, the mean sICAM-I levels in grade 1 oocytes were significantly decreased when compared to grade 2 and 3 oocytes.The reduction of the number of fertilized oocytes and transferred embryos represents the main target of assisted reproductive medicine. We propose sICAM-1 as a biochemical marker for oocyte maturation and grading, with a possible interesting rebound in assisted reproduction techniques.

  4. Association between induced abortion history and later in vitro fertilization outcomes.

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    Wang, Yao; Sun, Yun; Di, Wen; Kuang, Yan-Ping; Xu, Bing

    2018-03-06

    To establish an effective and safe clinical fertility strategy by investigating the relationship between abortion history and pregnancy outcomes of in vitro fertilization (IVF) treatment. In the present retrospective cohort study, data from IVF treatment cycles performed at a reproductive center in China between October 1, 2014, and October 31, 2015, were assessed. Outcomes were compared between women with a history of induced abortion and those without. There were 1532 IVF treatment cycles included; 454 patients had a history of induced abortion and 1078 did not. The spontaneous abortion rate was significantly higher (30/170 [17.6%] vs 41/420 [9.8%]; P=0.002) and the endometrium was significantly thinner (8.8±1.8 vs 9.7±1.8 cm; P=0.001) among patients with a history of induced abortion compared with those without. In a subgroup analysis of patients with a history of induced abortion, women who had undergone surgical abortions had a lower live delivery rate compared with medical abortions (29/76 [38%] vs 101/378 [27%]; P=0.039). Further, women who had a history of more than two surgical abortions had lower live delivery and clinical pregnancy rates (both Pabortion was associated with worse IVF outcomes, especially a history of more than two surgical abortions. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  5. Toxic trace metals and human oocytes during in vitro fertilization (IVF)

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    Bloom, Michael S.; Parsons, Patrick J.; Steuerwald, Amy J.; Schisterman, Enrique F.; Browne, Richard W.; Kim, Keewan; Coccaro, Gregory A.; Conti, Giulia C.; Narayan, Natasha; Fujimoto, Victor Y.

    2010-01-01

    Trace exposures to the toxic metals mercury (Hg), cadmium (Cd) and lead (Pb) may threaten human reproductive health. The aim of this study is to generate biologically-plausible hypotheses concerning associations between Hg, Cd, and Pb and in vitro fertilization (IVF) endpoints. For 15 female IVF patients, a multivariable log-binomial model suggests a 75% reduction in the probability for a retrieved oocyte to be in metaphase-II arrest for each μg/dL increase in blood Pb concentration (relative risk (RR) = 0.25, 95% confidence interval (CI) 0.03–2.50, P = 0.240). For 15 male IVF partners, each μg/L increase in urine Cd concentration is associated with an 81% decrease in the probability for oocyte fertilization (RR = 0.19, 95% CI 0.03–1.35, P = 0.097). Because of the magnitude of the effects, these results warrant a comprehensive study with sufficient statistical power to further evaluate these hypotheses. PMID:20096775

  6. Daily workload in the embryology laboratory and in vitro fertilization results.

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    Expósito, Antonia; Matorras, Roberto; Mendoza, Rosario; Crisol, Lorena; Martínez-Astorquiza, Txanton; Prieto, Begoña

    2010-01-01

    To ascertain if the daily activity in the in vitro fertilization (IVF) laboratory is related to pregnancy rates (PR) and fertilization rates (FR) in an IVF program. A retrospective study was performed to compare the PR and the FR obtained in 845 oocyte retrievals (OR) and 713 embryo transfers (ET), according to the daily workload. Different cutoffs were established: 3 OR per day; 3 ET per day, and also a cutoff considering the global activity in 3 different categories: optimal (level I), overload (level II) and high overload (level III), both the day of OR and of ET. The PR on the days with 3 OR were similar, as were the days with 3 ET. There were no differences in PR when the activity the day of OR was level I, II or III (24.4%, 25.2% and 28.3%, respectively) or when the activity on the day of ET was level I, II or III (29.6%, 37.3% and 23.7%, respectively). We failed to show any adverse results on our IVF program associated with the daily workload.

  7. Sperm DNA damage has a negative effect on early embryonic development following in vitro fertilization

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    Wei-Wei Zheng

    2018-01-01

    Full Text Available Sperm DNA damage is recognized as an important biomarker of male infertility. To investigate this, sperm DNA damage was assessed by the sperm chromatin dispersion (SCD test in semen and motile spermatozoa harvested by combined density gradient centrifugation (DGC and swim-up in 161 couples undergoing in vitro fertilization (IVF. Semen analysis and sperm DNA damage results were compared between couples who did or did not achieve pregnancy. The sperm DNA damage level was significantly different between the two groups (P < 0.05 and was negatively correlated with IVF outcomes. Logistic regression analysis confirmed that it was an independent predictor for achieving clinical pregnancy. The effects of different levels of sperm DNA damage on IVF outcomes were also compared. There were significant differences in day 3 embryo quality, blastocyst formation rate, and implantation and pregnancy rates (P < 0.05, but not in the basic fertilization rate between the two groups. Thus, sperm DNA damage as measured by the SCD appears useful for predicting the clinical pregnancy rate following IVF.

  8. Thyroid function during controlled ovarian hyperstimulation as part of in vitro fertilization.

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    Gracia, Clarisa R; Morse, Christopher B; Chan, Grace; Schilling, Samantha; Prewitt, Maureen; Sammel, Mary D; Mandel, Susan J

    2012-03-01

    To determine the exact nature and timing of alterations in thyroid function throughout controlled ovarian hyperstimulation (COH). Prospective cohort study. University fertility clinic. Fifty-seven women undergoing COH as part of planned in vitro fertilization. None. Timing and magnitude of change in serum thyroid hormones, including TSH, total and free T(4), E(2), and thyroxine-binding globulin (TBG), measured at six time points from before stimulation to 2 weeks after serum pregnancy test. Geometric mean serum TSH increased during stimulation, peaking 1 week after hCG administration compared with baseline (2.44 vs. 1.42 mIU/L), as did free T(4) (1.52 vs. 1.38 ng/dL) and TBG (32.86 vs. 21.52 μg/mL). Estradiol levels increased, peaking at hCG administration (1743.21 vs. 71.37 pg/mL). Of 50 women with baseline TSH ≤ 2.5 mIU/L, 22 (44.0%) had a subsequent rise in TSH to >2.5 during or after COH. The pattern of change over time in TSH concentrations was significantly influenced by baseline hypothyroidism and whether pregnancy was achieved. COH led to significant elevations in TSH, often above pregnancy appropriate targets. These findings were particularly evident in women with preexisting hypothyroidism and may have important clinical implications for screening and thyroid hormone supplementation. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  9. Factors influencing the success of in vitro fertilization for alleviating human infertility.

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    Edwards, R G; Fishel, S B; Cohen, J; Fehilly, C B; Purdy, J M; Slater, J M; Steptoe, P C; Webster, J M

    1984-03-01

    The program for in vitro fertilization at Bourn Hall began in October 1980. Various types of infertility have been treated during this time using the natural menstrual cycle or stimulation of follicular growth with antiestrogens and gonadotrophins. Follicular growth and maturation are assayed by urinary estrogens and LH, monitored regularly during the later follicular stage. Many patients had an endogenous LH surge; others needed an injection of HCG to induce ovulation. All oocytes were recovered by laparoscopy. Wide variations occurred in the time interval between the start of the LH surge and oocyte recovery and between oocyte recovery and insemination. Embryos taken between the one- and the eight-cell stage were replaced into their mother, no standard procedure being adopted for all patients. The results of all treatments including patient's responses during the follicular and luteal phases, oocyte recovery, fertilization, cleavage, replacement, implantation, abortion, and birth and the effect of factors such as replacing two or more embryos, maternal age, and previous obstetric history are described in detail. The incidence of implantation after embryo replacement improved from 16.5% initially to 30% currently. More than 118 babies have been born, and many pregnancies are continuing.

  10. Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

    Science.gov (United States)

    Bouillon, Céline; Léandri, Roger; Desch, Laurent; Ernst, Alexandra; Bruno, Céline; Cerf, Charline; Chiron, Alexandra; Souchay, Céline; Burguet, Antoine; Jimenez, Clément; Sagot, Paul; Fauque, Patricia

    2016-01-01

    In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371) were randomized according to two media (Single Step Medium--SSM group) or Global medium (Global group). This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73) conceived in the randomized study were included (42 for Global and 31 for SSM). The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded). The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items) were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (pculture medium had no significant effect on birthweight, risk of malformation (minor and major), growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002), irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.

  11. Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

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    Céline Bouillon

    Full Text Available In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371 were randomized according to two media (Single Step Medium--SSM group or Global medium (Global group. This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73 conceived in the randomized study were included (42 for Global and 31 for SSM. The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded. The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (p<0.05. The culture medium had no significant effect on birthweight, risk of malformation (minor and major, growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002, irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.

  12. Use of Bayesian Inference to Correlate In Vitro Embryo Production and In Vivo Fertility in Zebu Bulls

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    Mateus José Sudano

    2011-01-01

    Full Text Available The objective of this experiment was to test in vitro embryo production (IVP as a tool to estimate fertility performance in zebu bulls using Bayesian inference statistics. Oocytes were matured and fertilized in vitro using sperm cells from three different Zebu bulls (V, T, and G. The three bulls presented similar results with regard to pronuclear formation and blastocyst formation rates. However, the cleavage rates were different between bulls. The estimated conception rates based on combined data of cleavage and blastocyst formation were very similar to the true conception rates observed for the same bulls after a fixed-time artificial insemination program. Moreover, even when we used cleavage rate data only or blastocyst formation data only, the estimated conception rates were still close to the true conception rates. We conclude that Bayesian inference is an effective statistical procedure to estimate in vivo bull fertility using data from IVP.

  13. INHIBITION OF IN VITRO FERTILIZATION IN THE HAMSTER BY ANTIBODIES RAISED AGAINST THE RAT SPERM PROTEIN SP22

    Science.gov (United States)

    INHIBITION OF IN VITRO FERTILIZATION IN THE HAMSTER BY ANTIBODIES RAISED AGAINST THE RAT SPERM PROTEIN SP22. SC Jeffay*, SD Perreault, KL Bobseine*, JE Welch*, GR Klinefelter, US EPA, Research Triangle Park, NC. SP22, a rat sperm membrane protein that is highly-correlated w...

  14. Through the smoke: Use of in vivo and in vitro cigarette smoking models to elucidate its effect on female fertility

    Energy Technology Data Exchange (ETDEWEB)

    Camlin, Nicole J. [School of Environment and Life Sciences, University of Newcastle, Callaghan, NSW 2308 (Australia); McLaughlin, Eileen A., E-mail: eileen.mclaughlin@newcastle.edu.au [School of Environment and Life Sciences, University of Newcastle, Callaghan, NSW 2308 (Australia); Holt, Janet E. [School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW 2308 (Australia)

    2014-12-15

    A finite number of oocytes are established within the mammalian ovary prior to birth to form a precious ovarian reserve. Damage to this limited pool of gametes by environmental factors such as cigarette smoke and its constituents therefore represents a significant risk to a woman's reproductive capacity. Although evidence from human studies to date implicates a detrimental effect of cigarette smoking on female fertility, these retrospective studies are limited and present conflicting results. In an effort to more clearly understand the effect of cigarette smoke, and its chemical constituents, on female fertility, a variety of in vivo and in vitro animal models have been developed. This article represents a systematic review of the literature regarding four of experimental model types: 1) direct exposure of ovarian cells and follicles to smoking constituents’ in vitro, 2) direct exposure of whole ovarian tissue with smoking constituents in vitro, 3) whole body exposure of animals to smoking constituents and 4) whole body exposure of animals to cigarette smoke. We summarise key findings and highlight the strengths and weaknesses of each model system, and link these to the molecular mechanisms identified in smoke-induced fertility changes. - Highlights: • In vivo exposure to individual cigarette smoke chemicals alters female fertility. • The use of in vitro models in determining molecular mechanisms • Whole cigarette smoke inhalation animal models negatively affect ovarian function.

  15. In vitro fertilization (IVF) using semi-defined culture conditions from low or high antral follicle count pubertal beef heifers

    Science.gov (United States)

    To compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers, AFC were determined on 106 heifers using transrectal ultrasonography. Ten heifers with the lowest AFC (avg. 13.2) and 10 heifers with the highest AFC (avg. 27.4) with ev...

  16. In vitro fertilization (IVF) from low or high antral follicle count pubertal beef heifers using semi-defined culture conditions

    Science.gov (United States)

    Antral follicle counts (AFC) vary among pubertal beef heifers. Our objective was to compare the in vitro maturation and fertilization of oocytes collected from low and high AFC heifers. Previously we reported results using serum-based IVF media and in this study report results using semi-defined m...

  17. An overview of studies on early development, cognition, and psychosocial well-being in children born after in vitro fertilization

    NARCIS (Netherlands)

    Wagenaar, K.; Huisman, J.; Cohen-Kettenis, P.T.; Delemarre-van d Waal, H.A.

    2008-01-01

    OBJECTIVE:: To examine whether sufficient research has been done and definite conclusions can be drawn on the psychological outcome of children born after in vitro fertilization (IVF), a review was performed of studies on early development, cognition, and psychosocial well-being in IVF children.

  18. Development and evaluation of a porcine in vitro colon organ culture technique.

    Science.gov (United States)

    Costa, Matheus O; Harding, John C S; Hill, Janet E

    2016-10-01

    The intestinal mucosa comprises a complex assemblage of specialized tissues that interact in numerous ways. In vitro cell culture models are generally focused on recreating a specific characteristic of this organ and do not account for the many interactions between the different tissues. In vitro organ culture (IVOC) methods offer a way to overcome these limitations, but prolonging cell viability is essential. This study aimed to determine the feasibility and optimal conditions for in vitro culture of swine colonic mucosa for use as an enteric pathogen infection model. Explants (n = 168) from commercial pigs (n = 12), aged 5 to 10 wk, were used to assess the impact of various culture protocols on explant viability. Explants were cultured for up to 5 d and formalin fixed at 24-h intervals. Following establishment of the culture protocol, explants (n = 208) from 13 pigs were evaluated at Day 0 and 5 of culture. Assessment of viability was based on histological changes (tissue architecture evaluated by H&E, immunostaining of cell proliferation marker Ki-67) and expression of genes encoding IL-1α, IL-8, TNF-α, IFN-γ, and e-cadherin. After 5 d in culture, 20% of explants displayed over 80% of epithelial coverage, whereas 31% of explants had more than 50% of their surface covered by columnar epithelium, and 81% had crypts but with a decreased number of Ki-67-positive cells when compared to Day 0. Notably, large variability in explant quality was observed between donor pigs. Best possible explants were obtained from the distal colon of pigs, processed immediately after euthanasia, cultured at the liquid-tissue-gas interface in media supplemented with a mixture of antibiotics and antifungals and an oxygen-rich gas mix.

  19. The in vivo developmental potential of porcine skin-derived progenitors and neural stem cells.

    Science.gov (United States)

    Zhao, Ming-Tao; Yang, Xiaoyu; Lee, Kiho; Mao, Jiude; Teson, Jennifer M; Whitworth, Kristin M; Samuel, Melissa S; Spate, Lee D; Murphy, Clifton N; Prather, Randall S

    2012-09-20

    Multipotent skin-derived progenitors (SKPs) can be traced back to embryonic neural crest cells and are able to differentiate into both neural and mesodermal progeny in vitro. Neural stem cells (NSCs) are capable of self-renewing and can contribute to neuron and glia in the nervous system. Recently, we derived porcine SKPs and NSCs from the same enhanced green fluorescent protein (EGFP) transgenic fetuses and demonstrated that SKPs could contribute to neural and mesodermal lineages in vivo. However, it remains unclear whether porcine SKPs and NSCs can generate ectoderm and mesoderm lineages or other germ layers in vivo. Embryonic chimeras are a well-established tool for investigating cell lineage determination and cell potency through normal embryonic development. Thus, the purpose of this study was to investigate the in vivo developmental potential of porcine SKPs and fetal brain-derived NSCs by chimera production. Porcine SKPs, NSCs, and fibroblasts were injected into precompact in vitro fertilized embryos (IVF) and then transferred into corresponding surrogates 24 h postinjection. We found that porcine SKPs could incorporate into the early embryos and contribute to various somatic tissues of the 3 germ layers in postnatal chimera, and especially have an endodermal potency. However, this developmental potential is compromised when they differentiate into fibroblasts. In addition, porcine NSCs fail to incorporate into host embryos and contribute to chimeric piglets. Therefore, neural crest-derived SKPs may represent a more primitive state than their counterpart neural stem cells in terms of their contributions to multiple cell lineages.

  20. In vitro cytocompatibility of porcine type I atelocollagen crosslinked by oxidized glycogen.

    Science.gov (United States)

    Rousseau, Cécile F; Gagnieu, Christian H

    2002-03-01

    Oxidized glycogen is used as collagen crosslinker to obtain materials with defined crosslinking degrees. These materials are characterized by their swelling ratio. calorimetric properties and the crosslinking level. Direct and indirect cytotoxicities of the materials obtained as sheets, are evaluated in vitro in cultures of human fibroblasts. The crosslinking degree depends on the ratio CHO glycogen/NH2 glycogen, but whatever this ratio (4.0, 2.0 or 0.4), an important percentage of the introduced CHO groups remains free and these groups are responsible for the cytotoxicity observed with the strongly crosslinked materials. This cytotoxicity appears in cell shape modification and in significant reduction of cell growth. Whatever the crosslinking degree, this toxicity can be suppressed by a single treatment with sodium borohydride, which reduced the remaining free CHO groups in OH functions and stabilizes the materials by a concomitant reduction of the crosslinking imine bonds. After reduction, all materials allow cellular adhesion and proliferation. This new crosslinking method of the collagen by the oxidized glycogen could be promising in the preparation of matrix for in vitro and in vivo tissue regeneration.

  1. Physiochemical properties and resorption progress of porcine skin-derived collagen membranes: In vitro and in vivo analysis

    National Research Council Canada - National Science Library

    AN, Yin-Zhe; KIM, You-Kyoung; LIM, Su-Min; HEO, Yeong-Ku; KWON, Mi-Kyung; CHA, Jae-Kook; LEE, Jung-Seok; JUNG, Ui-Won; CHOI, Seong-Ho

    2018-01-01

    The aim of the present study was to evaluate the physiochemical properties and resorption progress of two cross-linked, porcine skin-derived collagen membranes and compare their features with those of...

  2. Mixed infections with Chlamydia and porcine epidemic diarrhea virus - a new in vitro model of chlamydial persistence

    Directory of Open Access Journals (Sweden)

    Kaiser Carmen

    2010-07-01

    Full Text Available Abstract Background Chlamydiae induce persistent infections, which have been associated with a wide range of chronic diseases in humans and animals. Mixed infections with Chlamydia and porcine epidemic diarrhea virus (PEDV may result in generation of persistent chlamydial infections. To test this hypothesis, an in vitro model of dual infection with cell culture-adapted PEDV and Chlamydia abortus or Chlamydia pecorum in Vero cells was established. Results Infected cultures were investigated by immunofluorescence (IF, transmission electron microscopy (TEM and re-infection experiments. By IF, Chlamydia-infected cells showed normal inclusions after 39 hpi. Dual infections with Chlamydia abortus revealed a heterogenous mix of inclusion types including small inclusions consisting of aberrant bodies (ABs, medium-sized inclusions consisting of ABs and reticulate bodies and normal inclusions. Only aberrant inclusions were observable in dual infection experiments with Chlamydia pecorum and PEDV. TEM examinations of mixed infections with Chlamydia abortus and Chlamydia pecorum revealed aberrant chlamydial inclusions containing reticulate-like, pleomorphic ABs, which were up to 2 μm in diameter. No re-differentiation into elementary bodies (EBs was detected. In re-infection experiments, co-infected cells produced fewer EBs than monoinfected cells. Conclusions In the present study we confirm that PEDV co-infection alters the developmental cycle of member species of the family Chlamydiaceae, in a similar manner to other well-described persistence induction methods. Interestingly, this effect appears to be partially species-specific as Chlamydia pecorum appears more sensitive to PEDV co-infection than Chlamydia abortus, as evidenced by TEM and IF observations of a homogenous population of aberrant inclusions in PEDV - Chlamydia pecorum co-infections.

  3. Sexually Dimorphic Effect of In Vitro Fertilization (IVF) on Adult Mouse Fat and Liver Metabolomes

    Science.gov (United States)

    Feuer, Sky K.; Donjacour, Annemarie; Simbulan, Rhodel K.; Lin, Wingka; Liu, Xiaowei; Maltepe, Emin

    2014-01-01

    The preimplantation embryo is particularly vulnerable to environmental perturbation, such that nutritional and in vitro stresses restricted exclusively to this stage may alter growth and affect long-term metabolic health. This is particularly relevant to the over 5 million children conceived by in vitro fertilization (IVF). We previously reported that even optimized IVF conditions reprogram mouse postnatal growth, fat deposition, and glucose homeostasis in a sexually dimorphic fashion. To more clearly interrogate the metabolic changes associated with IVF in adulthood, we used nontargeted mass spectrometry to globally profile adult IVF- and in vivo-conceived liver and gonadal adipose tissues. There was a sex- and tissue-specific effect of IVF on adult metabolite signatures indicative of metabolic reprogramming and oxidative stress and reflective of the observed phenotypes. Additionally, we observed a striking effect of IVF on adult sexual dimorphism. Male-female differences in metabolite concentration were exaggerated in hepatic IVF tissue and significantly reduced in IVF adipose tissue, with the majority of changes affecting amino acid and lipid metabolites. We also observed female-specific changes in markers of oxidative stress and adipogenesis, including reduced glutathione, cysteine glutathione disulfide, ophthalmate, urate, and corticosterone. In summary, embryo manipulation and early developmental experiences can affect adult patterns of sexual dimorphism and metabolic physiology. PMID:25211591

  4. Beta2 toxin is not involved in in vitro cell cytotoxicity caused by human and porcine cpb2-harbouring Clostridium perfringens.

    Science.gov (United States)

    Allaart, Janneke G; van Asten, Alphons J A M; Vernooij, Johannes C M; Gröne, Andrea

    2014-06-25

    Clostridium perfringens is a common cause of intestinal disease in animals and humans. Its pathogenicity is attributed to the toxins it can produce, including the beta2 toxin. The presence of cpb2, the gene encoding the beta2 toxin, has been associated with diarrhoea in neonatal piglets and humans. However, the exact role of the beta2 toxin in the development of diarrhoea is still unknown. In this study we investigated the level of cytotoxicity to porcine IPI-21 and human Caco-2 cell-lines caused by porcine and human cpb2-harbouring C. perfringens and the significance of the beta2 toxin for the induction of cell cytotoxicity. Supernatants of porcine cpb2-harbouring C. perfringens strains were cytotoxic to both cell lines. Cell cytotoxicity caused by supernatant of human cpb2-harbouring C. perfringens strains was variable among strains. However, removal of the beta2 toxin by anti-beta2 toxin antibodies or degradation of the beta2 toxin by trypsin did not reduce the cytotoxic effect of any of the supernatants. These data suggest that beta2 toxin does not play a role in the development of cell cytotoxicity in in vitro experiments. In vivo studies are necessary to definitely define the role of beta2 toxin in the development of cell cytotoxicity and subsequent diarrhoea. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Effect of co-culture human endothelial progenitor cells with porcine oocytes during maturation subsequent embryo development of parthenotes in vitro.

    Science.gov (United States)

    Lee, Seok Hee; Oh, Hyun Ju; Kim, Min Jung; Setyawan, Erif Maha Nugraha; Choi, Yoo Bin; Lee, Byeong Chun

    2018-02-14

    Human endothelial progenitor cells (EPCs) have been applied to regenerative medicine for their roles in angiogenesis as well as neovascularization, and these angiogenetic functions have beneficial effects on maturation of ovarian follicles. However, little information is available on whether EPCs on culture systems affect oocyte maturation and subsequent embryo development. Therefore, the objective of this study was to investigate the effect of EPC co-culture on porcine oocytes during in vitro maturation (IVM) and subsequent embryo development, and to examine gene expression in cumulus cells, oocytes and blastocysts. The effect of co-culture using EPC on porcine oocyte IVM was investigated. Oocytes were activated using electrical stimulation and embryo developmental competence was estimated. The expression of the genes related to cumulus expansion, oocyte maturation, embryo development and apoptosis were analyzed. In result, there was a significantly increased maturation rate in EPC group compared with control (p cultured with EPCs exhibited significantly improved blastocyst formation rates (p culture group showed significantly higher SOX2, OCT4 and NANOG levels. In conclusion, co-culturing porcine oocytes with EPCs improves their maturation by regulating genes involved in cumulus cell expansion, oocyte maturation and apoptosis. Moreover, EPC co-culture during IVM enhanced embryo development as shown by increased blastocyst formation rate and pluripotency-related gene expression. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  6. The effect of cumulus cells on domestic cat (Felis catus) oocytes during in vitro maturation and fertilization.

    Science.gov (United States)

    Sowińska, N; Frankowska, K; Filipczyk, A; Adamaszek, A; Nalik, K; Fic, K; Pietsch-Fulbiszewska, A

    2017-04-01

    The aim of this study was to evaluate the effect of co-culture of denuded oocytes with cumulus cells (CC) or cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) and in vitro fertilization (IVF). Immature oocytes were collected from ovaries of domestic cats following a routine ovariectomy. Oocytes were matured in vitro for 24 hr within four groups: (i) denuded oocytes (DO), (ii) DO co-cultured with CC, (iii) DO co-cultured with COC and (iv) COC as a control group. In further experiments, COCs were matured in vitro for 24 hr, and then, oocytes were randomly divided into four groups as previously described and fertilized in vitro. Embryos were cultured for up to 7 days. At the end of each experiment, oocytes/embryos were stained with Hoechst 33342 solution and observed under an inverted fluorescence microscope. The results of oocyte maturation showed that their meiotic competence decreased significantly in all experimental groups, compared to the control group. The maturation rates were approximately 45%, 24%, 43% and 76% in experiment 1, and 21%, 14%, 33% and 50% in experiment 2 in groups (i), (ii), (iii) and (iv), respectively. Examination of in vitro fertilization revealed that embryos developed up to the morula stage in all experimental groups. DO and oocytes cultured with COC during fertilization showed a lower cleavage rate-36% and 25% as opposed to those co-cultured with loose CC and the control group-43% and 42%, respectively. Results of this study indicate that cumulus cells connected with an oocyte into a cumulus-oocyte complex are irreplaceable for the maturation of domestic cat oocyte, but that the addition of loose CC may be beneficial for IVF. © 2016 Blackwell Verlag GmbH.

  7. Purification and characterization of porcine liver DNA polymerase gamma: utilization of dUTP and dTTP during in vitro DNA synthesis.

    OpenAIRE

    Mosbaugh, D W

    1988-01-01

    Porcine liver DNA polymerase gamma has been demonstrated to preferentially incorporate dTMP over dUMP during in vitro DNA synthesis. When polymerase activity was measured in standard reactions containing saturating levels of either dTTP or dUTP, the polymerization rate was slightly faster in the reaction containing dTTP. However, under conditions where both dTTP and dUTP competed, at an equal molar concentration, approximately 3-times more thymine residues were incorporated than uracil residu...

  8. In vitro fertilization as a diagnostic and therapeutic tool in a patient with partial 17,20-desmolase deficiency.

    Science.gov (United States)

    Pellicer, A; Miró, F; Sampaio, M; Gómez, E; Bonilla-Musoles, F M

    1991-05-01

    To present a case with 17,20-desmolase activity deficiency in which in vitro fertilization (IVF) served not only as a therapeutic approach but also as a diagnostic tool for the specificity of the enzymatic deficiency. IVF in the patient under study compared with a control group. All women treated with pure follicle-stimulating hormone (FSH). IVF program at the Instituto Valenciano de Infertilidad. A patient with primary amenorrhea, who was the subject under study, and seven normally cycling control patients undergoing IVF in the same series. IVF, steroidogenesis in vitro of granulosa-luteal cell obtained at ovum pick-up. Oocyte fertilization and embryo cleavage. Serum and follicular fluid (FF) levels of estradiol (E2), progesterone (P), testosterone (T), androstendione (A), 17 alpha-hydroxyprogesterone (17-OHP). In vitro accumulation of E2 and P. Ovulation induction with FSH was successful in achieving follicular development despite low circulating E2. Fertilization and cleavage rates were similar to the control subjects. The patient developed ovarian hyperstimulation. The lack of 17,20-desmolase activity was detected by normal P levels in serum and FF, high 17-OHP, and low T, A, and E2 levels in serum and FF. Granulosaluteal cell cultures in the presence of T restored normal E2 and P production in response to gonadotropins. In patients with 17,20-desmolase deficiency, follicular development, oocyte maturation, and fertilization can take place in a low estrogenic environment.

  9. Formation of reactive aldehydes (MDA, HHE, HNE) during the digestion of cod liver oil: comparison of human and porcine in vitro digestion models.

    Science.gov (United States)

    Tullberg, Cecilia; Larsson, Karin; Carlsson, Nils-Gunnar; Comi, Irene; Scheers, Nathalie; Vegarud, Gerd; Undeland, Ingrid

    2016-03-01

    In this work, we investigated lipid oxidation of cod liver oil during gastrointestinal (GI) digestion using two types of in vitro digestion models. In the first type of model, we used human GI juices, while we used digestive enzymes and bile from porcine origin in the second type of model. Human and porcine models were matched with respect to factors important for lipolysis, using a standardized digestion protocol. The digests were analysed for reactive oxidation products: malondialdehyde (MDA), 4-hydroxy-trans-2-nonenal (HNE), and 4-hydroxy-trans-2-hexenal (HHE) by liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS), and for free fatty acids (FFA) obtained during the digestion by gas chromatography-mass spectrometry (GC-MS). The formation of the oxidation products MDA, HHE, and HNE was low during the gastric digestion, however, it increased during the duodenal digestion. The formation of the oxidation products reached higher levels when digestive juices of human origin were used (60 μM of MDA, 0.96 μM of HHE, and 1.6 μM of HNE) compared to when using enzymes and bile of porcine origin (9.8, and 0.36 μM of MDA; 0.16, and 0.026 μM of HHE; 0.23, and 0.005 μM of HNE, respectively, in porcine models I and II). In all models, FFA release was only detected during the intestinal step, and reached up to 31% of total fatty acids (FA). The findings in this work may be of importance when designing oxidation oriented lipid digestion studies.

  10. Preparation of Extracellular Matrix Developed Using Porcine Articular Cartilage and In Vitro Feasibility Study of Porcine Articular Cartilage as an Anti-Adhesive Film

    Directory of Open Access Journals (Sweden)

    Ji Hye Baek

    2016-01-01

    Full Text Available In this study, we examined whether porcine articular cartilage (PAC is a suitable and effective anti-adhesive material. PAC, which contained no non-collagenous tissue components, was collected by mechanical manipulation and decellularization of porcine knee cartilage. The PAC film for use as an anti-adhesive barrier was easily shaped into various sizes using homemade silicone molds. The PAC film was cross-linked to study the usefulness of the anti-adhesive barrier shape. The cross-linked PAC (Cx-PAC film showed more stable physical properties over extended periods compared to uncross-linked PAC (UnCx-PAC film. To control the mechanical properties, Cx-PAC film was thermally treated at 45 °C or 65 °C followed by incubation at room temperature. The Cx-PAC films exhibited varying enthalpies, ultimate tensile strength values, and contact angles before and after thermal treatment and after incubation at room temperature. Next, to examine the anti-adhesive properties, human umbilical vein endothelial cells (HUVECs were cultured on Cx-PAC and thermal-treated Cx-PAC films. Scanning electron microscopy, fluorescence, and MTT assays showed that HUVECs were well adhered to the surface of the plate and proliferated, indicating no inhibition of the attachment and proliferation of HUVECs. In contrast, Cx-PAC and thermal-treated Cx-PAC exhibited little and/or no cell attachment and proliferation because of the inhibition effect on HUVECs. In conclusion, we successfully developed a Cx-PAC film with controllable mechanical properties that can be used as an anti-adhesive barrier.

  11. In vitro induction of the acrosome reaction in bull sperm and the relationship to field fertility using low-dose inseminations

    DEFF Research Database (Denmark)

    Birck, A; Christensen, P; Labouriau, R

    2010-01-01

    The acrosome reaction (AR) is a prerequisite for normal sperm fertilizing capability and can be studied in vitro after induction by various agents. The efficacy of a sperm population to undergo the AR in vivo is expected to influence male fertilizing potential. During the past two decades, a number...... of attempts have been made to relate the in vitro-induced AR to field fertility in several species. However, to our knowledge, no studies have combined in vitro induction of the AR with the simultaneous detection of sperm viability and acrosomal status using a high-precision flow cytometric technique....... Furthermore, large-scale fertility trials using low-dose inseminations are pending. In the current study, the relationship between field fertility and the in vitro-induced AR was investigated using three ejaculates from each of 195 bulls, 156 Holstein and 39 Jersey bulls (Bos taurus), participating...

  12. Renal and Hepatic Functions after A Week of Controlled Ovarian Hyperstimulation during In Vitro Fertilization Cycles

    Directory of Open Access Journals (Sweden)

    Romito Ilaria

    2017-01-01

    Full Text Available Background: One the main aspects of in vitro fertilization (IVF cycle is to avoid any possible systemic damage on women undergoing a controlled ovarian hyperstimulation (COH. The aim of this work is to evaluate renal and hepatic function blood tests in patients undergoing controlled ovarian hyperstimulation during IVF cycles. Materials and Methods: We performed a prospective cohort analysis. All patients re- ceived a long stimulation protocol with gonadotropin-releasing hormone (GnRH analogues by daily administration, since the twenty-first day of the previous ovarian cycle followed by COH with recombinant follicle-stimulating hormone (FSH. The daily dose of exogenous gonadotropins for every single patient was modified according to her follicular growth. The oocytes were retrieved during the oocyte pick up and fertilized by standard procedures of intracytoplasmic sperm injection (ICSI. The blood samples to evaluate renal and hepatic functions were taken at the 7th day of ovarian stimulation. Results: We enrolled 426 women aged between 19 and 44 years, with a mean body mass index (BMI of 24.68 Kg/m2. The mean value of blood urea nitrogen was 14 ± 3.16 mg/ dl, creatinine: 1 ± 0.45 mg/dl, uric acid: 4 ± 1.95 mg/dl, total proteins: 7 ± 3.93 mg/dl, aspartate aminotransferase: 18 ± 6.29 mU/ml, alanine aminotransferase: 19 ± 10.41 mU/ ml, alkaline phosphatase: 81 ± 45.25 mU/ml, total bilirubin 1 ± 0.35 mg/dL. All of the results were considered as a normal range following the Medical Council of Canada. Conclusion: Our data suggest that, unlike ovarian hyperstimulation syndrome (OHSS, COH patients did not show any alteration to renal and hepatic functions.

  13. In vitro fertilization – from concept to first child Commemorating the 2010 Nobel Prize in Medicine

    Directory of Open Access Journals (Sweden)

    Kristina Janežič

    2011-09-01

    Full Text Available Since the birth of the world’s first test-tube baby in 1978, more than 4 million babies have been born worldwide with the help of in vitro fertilization (IVF. In Slovenia, the first babies were born in 1984. In 2005, the percentage of babies born with biomedically assisted reproduction was 3.9 % and was the highest in Europe. IVF is nowadays a widely available and accepted method for helping infertile couples. However, the path to success was long and difficult. In this article, we present the development of IVF from the first ideas about fertilizing eggs outside the body from the end of the 19th century, followed by experiments on different animal species, the first human pregnancy and finally the birth of the first child in the United Kingdom. Many pioneers from other countries, particularly Australia and the United States of America, were instrumental in developing IVF as we know it today. Unfortunately, covering the history of IVF would greatly exceed the purpose of this article. During its history, the method was modified with innovative improvements. In its beginnings, there was a great deal of scepticism and opposition from certain public as well as scientific circles, an attitude that changed radically. The fact that the pioneer Robert Edwards received the Nobel Prize for Medicine in 2010 is proof of the growing acceptance of the method. Surely, IVF is a complex medical field, which raised and still raises ethical and legal questions. But from a merely human perspective, it is simply a method which gave infertile couples the opportunity to fulfill their lifetime dream.

  14. Effect of body mass index on in vitro fertilization outcomes in women

    Directory of Open Access Journals (Sweden)

    Anjali Sathya

    2010-01-01

    Full Text Available Background :Obesity has become a major health problem across the world. In women, it is known to cause anovulation, subfecundity, increased risk of fetal anomalies and miscarriage rates. However, in women going for assisted reproduction the effects of obesity on egg quality, embryo quality, clinical pregnancy, live birth rates are controversial. Objectives :To assess the effect of women′s body mass index (BMI on the reproductive outcome of non donor In vitro fertilization (IVF/Intracytoplasmic sperm injection (ICSI. The effects of BMI on their gonadotrophin levels (day 2 LH, FSH, gonadotrophin dose required for ovarian stimulation, endometrial thickness and oocyte/embryo quality were looked at, after correcting for age and poor ovarian reserve. Materials and Methods : Retrospective study of medical records of 308 women undergoing non donor IVF cycles in a University affiliated teaching hospital. They were classified into three groups: normal weight (BMI25 30 kg/m 2 . All women underwent controlled ovarian hyper stimulation using long agonist protocol. Results : There were 88 (28.6% in the normal weight group, 147 (47.7% in the overweight and 73 (23.7% in the obese group. All three groups were comparable with respect to age, duration of infertility, female and male causes of infertility. The three groups were similar with respect to day 2 LH/FSH levels, endometrial thickness and gonadotrophin requirements, oocyte quality, fertilization, cleavage rates, number of good quality embryos and clinical pregnancy rates. Conclusion :Increase in body mass index in women does not appear to have an adverse effect on IVF outcome. However, preconceptual counselling for obese women is a must as weight reduction helps in reducing pregnancy-related complications.

  15. Alcohol consumption and quality of embryos obtained in programmes of in vitro fertilization

    Directory of Open Access Journals (Sweden)

    Artur Wdowiak

    2014-06-01

    Full Text Available introduction. Infertility is defined as a state when a couple fails to conceive a pregnancy after one year of regular intercourse without the use of contraception. Alcohol consumption is one of the main stimulants which negatively affect the female and male reproductive system. objective. The objective of the study was analysis of the effect of alcohol consumption by the examined women on the quality of embryos obtained during in vitro fertilization programmes. material and methods. The study covered 54 women who received treatment due to infertility. The database and statistical analyses were performed using computer software STATISTICA 7.1 (StatSoft, Poland. results. The study showed that 42.59% from among 100% of the women in the study consumed alcohol. In the group of women who consumed alcohol, class A embryos constituted 4.35%, class B embryos – 86.96%, while embryos of class C – 8.69%. A statistically significant difference was observed between the classes of embryos and alcohol consumption by the women examined (p=0.001. In addition, a statistically significant relationship was found between the amount of alcohol consumed and the classes of embryos (p=0.005. A significantly larger number of class B embryos came from women who consumed more than 25 grams of ethyl alcohol daily (72.72%, compared to those who consumed alcohol sporadically (44.44%, or those who abstained entirely from alcohol (30.00%. conclusions. Alcohol consumption causes the development of poorer quality embryos. Significantly more embryos of class B came from oocytes of women who consumed alcohol, compared to class A. An active campaign against alcohol consumption should be carried out among women at reproductive age to safeguard their fertility and future motherhood.

  16. The in vitro comparative cytopathology of a porcine rotavirus and the simian prototype (SA-11

    Directory of Open Access Journals (Sweden)

    Lonien S.C.H.

    2001-01-01

    Full Text Available A citopatologia in vitro de uma cepa de rotavírus porcino adaptado em cultura de células foi comparada à estirpe-protótipo símia (SA-11. O efeito citopático (ECP produzido pelos vírus foi semelhante embora a estirpe porcina tivesse apresentado algumas alterações diferentes, como o acentuado estreitamento do citoplasma, com grande perda do volume citoplasmático. O vírus porcino apresentou menor número de plaques de ECP porém com diâmetro maior em relação ao vírus símio, demonstrando maior capacidade de disseminação célula-célula, quase oito vezes mais, a julgar pelo diâmetro dos plaques de ECP. Os elementos do citoesqueleto das células infectadas revelaram uma reorganização semelhante para ambas as estirpes, não sendo possível observar nenhuma diferença, embora o ECP do vírus porcino tenha sido mais acentuado.

  17. Male caffeine and alcohol intake in relation to semen parameters and in vitro fertilization outcomes among fertility patients.

    Science.gov (United States)

    Karmon, A E; Toth, T L; Chiu, Y-H; Gaskins, A J; Tanrikut, C; Wright, D L; Hauser, R; Chavarro, J E

    2017-03-01

    Much of the literature on the impact of male caffeine and alcohol intake on reproductive outcomes has utilized semen quality as a proxy for male fertility, although semen parameters have a limited predictive value for spontaneous pregnancy. The objective of this study was to investigate whether male caffeine and alcohol intakes are associated with semen parameters and assisted reproductive technology outcome. The Environment and Reproductive Health Study, an ongoing prospective cohort study, enrolls subfertile couples presenting for treatment at an academic fertility center (2007-2012). A total of 171 men with 338 semen analyses and 205 assisted reproductive technology cycles were included in this analysis. Diet was assessed using a 131-item food frequency questionnaire. Mixed models adjusting for potential confounders were used to evaluate the relationships of male caffeine and alcohol intakes with semen parameters and assisted reproductive technology outcomes. There was no association between male caffeine and alcohol intake and semen quality. Male caffeine intake was negatively related to live birth after assisted reproductive technologies (p-trend caffeine intake (≥272 mg/day) compared to couples with a male partner in the lowest quartile of intake (caffeine and alcohol intakes were associated with live birth after assisted reproductive technologies, but not with semen parameters, among fertility patients. © 2017 American Society of Andrology and European Academy of Andrology.

  18. 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.

  19. In vitro oocyte maturation, fertilization and culture after ovum pick-up in an endangered gazelle (Gazella dama mhorr).

    Science.gov (United States)

    Berlinguer, F; González, R; Succu, S; del Olmo, A; Garde, J J; Espeso, G; Gomendio, M; Ledda, S; Roldan, E R S

    2008-02-01

    The recovery of immature oocytes followed by in vitro maturation, fertilization and culture (IVMFC) allows the rescue of biological material of great genetic value for the establishment of genetic resource banks of endangered species. Studies exist on sperm cryopreservation of endangered Mohor gazelle (Gazella dama mhorr), but no work has been carried out yet on oocyte collection, fertilization and culture in this or related species. The purpose of this study was to develop a protocol for ovarian stimulation for the recovery of oocytes and subsequent IVMFC in the Mohor gazelle using frozen-thawed spermatozoa. Ovum pick-up was performed after ovarian stimulation with a total dose of 5.28 mg of ovine FSH. A total of 35 oocytes were recovered from 56 punctured follicles (62%) (N=6 females). Out of 29 cumulus-oocyte complexes matured in vitro, 3% were found at germinal vesicle stage, 7% at metaphase I, 21% were degenerated, and 69% advanced to metaphase II. Fertilization and cleavage rates of matured oocytes were 40 and 30%, respectively. Embryos cleaved in vitro up to the 6-8 cell stage but none progressed to the blastocyst stage, suggesting the existence of a developmental block and the need to improve culture conditions. Although more studies are needed to improve hormonal stimulation and oocyte harvesting, as well as IVMFC conditions, this study demonstrates for the first time the feasibility of in vitro fertilization with frozen-thawed semen of in vitro matured oocytes collected by ovum pick-up from FSH-stimulated endangered gazelles.

  20. Efeito do pH na calcificação in vitro de pele porcina In Vitro calcification of porcine skin: influence of pH

    Directory of Open Access Journals (Sweden)

    Thelma M. Batista

    2007-12-01

    Full Text Available A engenharia de tecidos tem sido utilizada como alternativa na reconstrução de tecidos duros e moles. Este estudo teve como objetivo a calcificação "in vitro" de pele porcina visando à obtenção de um material para regeneração de tecido duro. As matrizes de pele porcina foram calcificadas em cela dupla termostatizada a 37 °C em pH 7,4 e pH 9,0 e caracterizadas por microscopia eletrônica de varredura (MEV, termogravimetria (TGA, espectroscopia no infravermelho (FTIR, calorimetria exploratória diferencial (DSC e difração de raios X. Os resultados obtidos por DSC mostraram que as amostras calcificadas têm um pequeno aumento nos valores de temperatura de desnaturação em relação à amostra não calcificada, enquanto as curvas termogravimétricas mostraram uma porcentagem maior de material inorgânico para o pH 7,4 em comparação com as amostras obtidas em pH 9,0. A formação de sais de fosfato de cálcio nas fibras de colágeno foi confirmada por difração de raios X (DRX, espectroscopia no infravermelho (FTIR e microscopia eletrônica de varredura (MEV.Tissue engineering has been used as an alternative in the reconstruction of hard and soft tissues. The objective of this work was to study the in vitro calcification of porcine skin aiming at obtaining a material to be used for regeneration of hard tissue. Pig skin was calcified at 37 °C, pH 7.4 and pH 9.0 and characterized by scanning electron microscopy (SEM, thermogravimetric analysis (TGA, infrared spectroscopy (FTIR, differential scanning calorimetry (DSC and X ray diffraction (XRD. The results obtained by DSC showed that calcified samples had a small increase in the values of denaturation temperature when compared with uncalcified samples. Thermogravimetric curves showed higher quantity of inorganic material for the pH 7.4 matrix when compared to samples obtained in pH 9.0. The formation of calcium phosphate salts on collagen fibers was seen by SEM, which was also confirmed

  1. Optimization of culture conditions for in vitro fertilization and reproduction of Microphallus turgidus (Trematoda: Microphallidae).

    Science.gov (United States)

    Pung, Oscar J; Lester, Terry; Burger, Ashley R; Alyanak, Efe; O'Leary, Patricia A

    2011-02-01

    Most attempts to culture adult digeneans in vitro are unsuccessful. Even progenetic digeneans typically fail to produce infective eggs in axenic culture. However, metacercariae of Microphallus turgidus grown in vitro mature into adults and release eggs infective to the hydrobiid snail Spurwinkia salsa. The objectives of the present study were to verify the reproducibility of the M. turgidus culture protocol, to define optimal culture conditions for M. turgidus further, and to investigate why the parasite can be grown successfully in the absence of the definitive host. In the original cultivation protocol, excysted M. turgidus metacercariae from grass shrimp (Palaemonetes pugio) were incubated overnight in a conical-bottom centrifuge tube containing Hank's balanced salt solution and then cultivated in flat-bottom culture plate wells containing RPMI-1640 plus 20% horse serum. The gas phase was air. Worms cultured under this protocol consistently deposited eggs infective to snails. Worms grown in anaerobic conditions deposited few eggs, and those cultured in a gas phase of 5% CO(2) survived longer and produced more eggs than those cultured in air. However, snails were less likely to become infected when fed eggs deposited by worms cultured in 5% CO(2). Additionally, worms incubated with conspecifics in conical-bottom tubes prior to cultivation were more likely to be inseminated than worms incubated in flat-bottom culture wells; the highest percentages of inseminated worms occurred when metacercariae were incubated 24 hr in conical-bottom tubes at a density of 50 worms/tube and at a temperature of 37 C. Worms incubated in the absence of conspecifics were not fertilized and failed to produce infective eggs.

  2. Significant Down-Regulation of “Biological Adhesion” Genes in Porcine Oocytes after IVM

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    Joanna Budna

    2017-12-01

    Full Text Available Proper maturation of the mammalian oocyte is a compound processes determining successful monospermic fertilization, however the number of fully mature porcine oocytes is still unsatisfactory. Since oocytes’ maturation and fertilization involve cellular adhesion and membranous contact, the aim was to investigate cell adhesion ontology group in porcine oocytes. The oocytes were collected from ovaries of 45 pubertal crossbred Landrace gilts and subjected to two BCB tests. After the first test, only granulosa cell-free BCB+ oocytes were directly exposed to microarray assays and RT-qPCR (“before IVM” group, or first in vitro matured and then if classified as BCB+ passed to molecular analyses (“after IVM” group. As a result, we have discovered substantial down-regulation of genes involved in adhesion processes, such as: organization of actin cytoskeleton, migration, proliferation, differentiation, apoptosis, survival or angiogenesis in porcine oocytes after IVM, compared to oocytes analyzed before IVM. In conclusion, we found that biological adhesion may be recognized as the process involved in porcine oocytes’ successful IVM. Down-regulation of genes included in this ontology group in immature oocytes after IVM points to their unique function in oocyte’s achievement of fully mature stages. Thus, results indicated new molecular markers involved in porcine oocyte IVM, displaying essential roles in biological adhesion processes.

  3. In vitro and in vivo effects of ulipristal acetate on fertilization and early embryo development in mice.

    Science.gov (United States)

    Gómez-Elías, Matías D; Munuce, María J; Bahamondes, Luis; Cuasnicú, Patricia S; Cohen, Débora J

    2016-01-01

    Does ulipristal acetate (UPA), a selective progesterone receptor modulator used for emergency contraception (EC), interfere with fertilization or early embryo development in vitro and in vivo? At doses similar to those used for EC, UPA does not affect mouse gamete transport, fertilization or embryo development. UPA acts as an emergency contraceptive mainly by inhibiting or delaying ovulation. However, there is little information regarding its effects on post-ovulatory events preceding implantation. This was an in vitro and in vivo experimental study involving the use of mouse gametes and embryos from at least three animals in each set of experiments. For in vitro fertilization experiments, mouse epididymal spermatozoa capacitated in the presence of different concentrations of UPA (0-1000 ng/ml) were used to inseminate cumulus-intact or cumulus-free eggs in the presence or absence of UPA during gamete co-incubation, and the percentage of fertilized eggs was determined. For in vivo fertilization experiments, superovulated females caged with proven fertile males were injected with UPA (40 mg/kg) or vehicle just before or just after mating and the percentage of fertilized eggs recovered from the ampulla was determined. To investigate the effect of UPA on embryo development, zygotes were recovered from mated females, cultured in the presence of UPA (1000 ng/ml) for 4 days and the progression of embryo development was monitored daily. In vitro studies revealed that the presence of UPA during capacitation and/or gamete co-incubation does not affect fertilization. Whereas the in vivo administration of UPA at the same time as hCG injection produced a decrease in the number of eggs ovulated compared with controls (vehicle injected animals, P < 0.05), no effects on fertilization were observed when UPA was administered shortly before or after mating. No differences were observed in either the percentage of cleaved embryos or the cleavage speed when UPA was present during in

  4. Increased time to pregnancy is associated with less optimal neurological condition in 4-year-old singletons, in vitro fertilization itself is not

    NARCIS (Netherlands)

    Schendelaar, P.; van den Heuvel, Edwin; Heineman, M. J.; La Bastide-Van Gemert, S.; Middelburg, K. J.; Seggers, Jorien; Hadders-Algra, M.

    2014-01-01

    STUDY QUESTION: Does ovarian hyperstimulation, the in vitro procedures required for in vitro fertilization (IVF)/intracytoplasmic sperm injection or the combination of both, affect the neurological outcome of 4-year-old singletons? SUMMARY ANSWER: Ovarian hyperstimulation, the in vitro procedure and

  5. Cryopreservation/transplantation of ovarian tissue and in vitro maturation of follicles and oocytes: Challenges for fertility preservation

    Directory of Open Access Journals (Sweden)

    Agarwal Ashok

    2008-10-01

    Full Text Available Abstract Cryopreservation of ovarian tissue and in vitro follicle maturation are two emerging techniques for fertility preservation, especially in cancer patients. These treatment regimes are opening up more options and allow for more suitable choices to preserve fertility according to the patient's specific circumstances. If these technologies are to become widely accepted, they need to be safe, easy to perform and must obtain favorable results. The generation of healthy eggs with the normal genetic complement and the ability to develop into viable and healthy embryos requires tight regulation of oocyte development and maturation. Novel freezing techniques such as vitrification, along with whole ovary cryopreservation and three-dimensional follicle cultures, have shown favorable outcomes. The scope of this article is to take a comprehensively look at the challenges still faced in order for these novel technologies to be routinely employed with the aim of successful fertility preservation.

  6. Support through patient internet-communities: Lived experience of Russian in vitro fertilization patients

    Directory of Open Access Journals (Sweden)

    Olga G. Isupova

    2011-07-01

    Full Text Available The article is concerned with the life experiences of infertile women going through infertility treatment and their need for social and psychological support, which they try to find in their immediate social environment. The Internet has become one place where everyone can find “people like oneself.” The best support is received from these people who are in the same life situation and are able and willing to share their lived experiences with each other. Communication via the Internet and the formation of a virtual community of patients has both positive and negative aspects, all of which are examined in the article. On the one hand, it creates a psychologically favorable atmosphere and might potentially increase the success rate of IVF treatment. On the other, this leads to the seclusion of patients within the circle of “similar people” and sometimes to negative attitudes towards people outside the circle. The article is based on the author's “netnography” research of a virtual community of Russian In-Vitro Fertilization (IVF1 patients.

  7. Survey of the Situation of Infertile Women Seeking In Vitro Fertilization Treatment in China

    Directory of Open Access Journals (Sweden)

    Xuan Jin

    2013-01-01

    Full Text Available Background. In previous studies, people’s knowledge of reproductive health and infertile women’s psychological states was surveyed in several countries. However, there has been limited information concerning the psychological states of infertile women seeking treatment and the outcomes of in vitro fertilization (IVF in China. Methods. Infertile women were asked to complete short questionnaires on the day that their oocytes were retrieved; these questionnaires covered the durations of their infertility, levels of education, sources of pressure, and psychological states. Data concerning IVF outcomes were provided by embryologists and clinicians. The correlations between the duration of infertility and educational level, psychological state and education level, and psychological state and outcome of IVF were analyzed in the cohort study. Results. The duration of infertility in more than half of the females was longer than 5 years. Compared with less-educated women, women with higher levels of education sought treatment earlier and their rates of depressive symptoms were lower. There is an association between negative emotions and outcome of IVF. Conclusions. The survey of the situations of infertile women seeking IVF treatment in China indicates the importance of popularizing knowledge concerning reproductive health. Improving medical conditions, reducing the costs of treatment, and developing social culture will aid in relieving the stress of infertile women and improving assisted reproductive treatment.

  8. The psychological profile and affective response of women diagnosed with unexplained infertility undergoing in vitro fertilization.

    Science.gov (United States)

    Aisenberg Romano, Gabi; Ravid, Hila; Zaig, Inbar; Schreiber, Shaul; Azem, Foad; Shachar, Izhak; Bloch, Miki

    2012-12-01

    It has been hypothesized that unexplained infertility may be related to specific personality and coping styles. We studied two groups of women with explained infertility (EIF, n = 63) and unexplained infertility (UIF, n = 42) undergoing an in vitro fertilization (IVF) cycle. Women completed personality and coping style questionnaires prior to the onset of the cycle, and state depression and anxiety scales before and at two additional time points during the cycle. Almost no in-between group differences were found at any of the measured time points in regards to the Minnesota Multiphasic Personality Inventory-2 validity and clinical scales, Illness Cognitions and Life Orientation Test, or for the situational measures. The few differences found suggest a more adaptive, better coping, and functioning defensive system in women with EIF. In conclusion, we did not find any clinically significant personality differences or differences in depression or anxiety levels between women with EIF and UIF during an IVF cycle. Minor differences found are probably a reaction to the ambiguous medical situation with its uncertain prognosis, amplifying certain traits which are not specific to one psychological structure but rather to the common experience shared by the group. The results of this study do not support the possibility that personality traits are involved in the pathophysiology of unexplained infertility.

  9. Increased incidence of ectopic pregnancy after in vitro fertilization in women with decreased ovarian reserve.

    Science.gov (United States)

    Lin, Shengli; Yang, Rui; Chi, Hongbin; Lian, Ying; Wang, Jiejing; Huang, Shuo; Lu, Cuiling; Liu, Ping; Qiao, Jie

    2017-02-28

    The incidence of ectopic pregnancy after assisted reproductive technology is increased approximately 2.5-5-fold compared with natural conceptions.Strategies were used to decrease the incidence of ectopic pregnancy, but ectopic pregnancy still occurs. In the present study, women were selected with decreased ovarian reserve (defined as FSH > 10 IU/L) aged 20 to 38 years who underwent IVF-ET between 2009 and 2014. These 2,061 women were age-matched with an equal number of women with normal ovarian reserve (defined as FSH ≤ 10 IU/L). During cycles following fresh embryo transfer, 93 patients were diagnosed with ectopic pregnancy. The incidence of ectopic pregnancy in clinical pregnancies was significantly higher in the decreased ovarian reserve than in the normal ovarian reserve group (5.51% vs. 2.99%). After adjusting for confounding factors, the incidence of ectopic pregnancy was significantly associated with decreased ovarian reserve. Our results showed that decreased ovarian reserve is an independent risk factor for ectopic pregnancy after in vitro fertilization-embryo transfer.

  10. Constitutive RB1 mutation in a child conceived by in vitro fertilization: implications for genetic counseling

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    Lucena Evandro

    2009-07-01

    Full Text Available Abstract Background The purpose of this study was to identify mutations associated with bilateral retinoblastoma in a quadruplet conceived by in vitro fertilization, and to trace the parental origin of mutations in the four quadruplets and their father. Methods Mutational screening was carried out by sequencing. Genotyping was carried out for determining quadruplet zygosity. Results The proband was a carrier of a novel RB1 constitutive mutation (g.2056C>G which was not detected in her father or her unaffected sisters, and of two other mutations (g.39606 C>T and g.174351T>A also present in two monozygotic sisters. The novel mutation probably occurred de novo while the others were of likely maternal origin. The novel mutation, affecting the Kozak consensus at the 5'UTR of RB1 and g.174351T>A were likely associated to retinoblastoma in the proband. Conclusion Molecular diagnosis of retinoblastoma requires genotypic data of the family for determining hereditary transmission. In the case of children generated by IVF with oocytes from an anonymous donor which had been stored in a cell repository, this might not be successfully accomplished, making precise diagnosis impracticable for genetic counseling.

  11. Prevalence of dental attrition in in vitro fertilization children of West Bengal.

    Science.gov (United States)

    Kar, Sudipta; Sarkar, Subrata; Mukherjee, Ananya

    2014-01-01

    Dental attrition is one of the problems affecting the tooth structure. It may affect both in vitro fertilization (IVF) and spontaneously conceived children. This study was aimed to evaluate and to compare the prevalence of dental attrition in deciduous dentition of IVF and spontaneously conceived children. In a cross-sectional case control study dental attrition status of 3-5 years old children were assessed. The case group consisted of term, singleton babies who were the outcome of IVF in the studied area in 2009. The control group consisted of term, first child, singleton and spontaneously conceived 3-5 years old children who were also resident of the studied area. A sample of 153 IVF and 153 spontaneously conceived children was examined according to Hansson and Nilner classification. Statistical analysis was carried out using Chi-square tests (χ(2) ) or Z test. No statistically significant difference found in studied (IVF children) and control group (spontaneously conceived children). IVF children are considered same as spontaneously conceived children when studied in relation to dental attrition status.

  12. Grief responses and coping strategies among infertile women after failed in vitro fertilization treatment.

    Science.gov (United States)

    Lee, Shu-Hsin; Wang, Shu-Chuan; Kuo, Ching-Pyng; Kuo, Pi-Chao; Lee, Maw-Sheng; Lee, Meng-Chih

    2010-09-01

    Reproductive technology has increased the childbearing potential for many infertile women, but in vitro fertilization (IVF) failures are common, which often trigger grief responses and coping strategies to manage the stressful life event. The present cross-sectional study investigated 66 women who had experienced at least one failure with IVF treatment. The data were gathered by a self-administered structured questionnaire, and included the participant's personal profile, grief responses and the Jalowiec's coping scale. The most common grief response among the respondents was bargaining, followed by acceptance, depression, anger, denial, and isolation. The order of coping strategies used, from highest-to-lowest, were confrontative, optimistic, self-reliant, fatalistic, supportive, evasive, palliative, and emotive. Use and self-perceived effectiveness among all coping strategies had a high correlation, except emotion. Bargaining, the most common grief response, was associated with a variety of coping strategies. All coping strategies were correlated with grief responses. The results of identifying the grief responses and associated coping strategies of women who have undergone failed IVF treatment may assist nurses and other health care professionals in their efforts to provide appropriate information, care and psychological support. © 2010 The Authors. Journal compilation © 2010 Nordic College of Caring Science.

  13. Bowel complications of deep endometriosis during pregnancy or in vitro fertilization.

    Science.gov (United States)

    Setúbal, António; Sidiropoulou, Zacharoula; Torgal, Mariana; Casal, Ester; Lourenço, Carlos; Koninckx, Philippe

    2014-02-01

    To review bowel complications caused by deep endometriosis during pregnancy or in vitro fertilization (IVF). Three case reports and a systematic review. A tertiary referral center for deep endometriosis surgery. Three case reports of bowel perforation or occlusion during pregnancy caused by deep endometriosis. A PubMed search was conducted to identify complications of deep endometriosis during pregnancy or IVF. The literature search identified 13 articles. According to these, 12 articles described 12 bowel complications caused by progression of deep endometriosis during pregnancy, and 1 article described six cases of bowel occlusion during IVF. In 12 of 15 women, complications occurred during the third trimester of pregnancy, whereas 3 of 15 women presented with complications in the postpartum period. All complications during IVF occurred during stimulation. No specific factors that could predict these complications were identified, leading to the conclusion that endometriosis complications that occur in pregnancy or in IVF patients are probably underreported. Bowel complications during pregnancy or IVF stimulation may occur in women with deep endometriosis. This suggests that the endocrine environment of pregnancy does not prevent progression, at least in some women. These complications are rare, although probably underreported. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  14. [Blood cell chimerism in dizygotic twins conceived by in vitro fertilization].

    Science.gov (United States)

    Martos-Moreno, G Á; Campos, C; Flores, R; Yturriaga, R; Pérez-Jurado, L A; Argente, J

    2013-10-01

    We present a case of hematopoietic chimerism in dizygotic twins (male and female) conceived by in vitro fertilization (IVF). At 8 years of age a blood karyotype was performed on the female due to the presence of clitoromegaly. Two different lines: 46,XX (53%) and 46,XY (47%) were found. FISH studies confirmed the presence of the SRY gene in 46,XY cells. Karyotyping of the male showed two different lines: 46,XY (58%) and 46,XX (42%). SRY gene was present in 46,XY cells. Microsatellite analyses of blood DNA revealed tetra-allelic contribution at some autosomal loci with similar proportions of maternal and paternal alleles and X/Y chromosome dose. FISH in buccal mucous showed that all cells from the female were 46,XX and those from the male 46,XY. The gonadal karyotype in the female was 46,XX without SRY. Hence, we report 46,XX/46,XY chimerism in dizygotic twins. Blood chimerism was confirmed by performing FISH on the buccal cells of the patients. Copyright © 2013 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

  15. Support through patient internet-communities: Lived experience of Russian in vitro fertilization patients.

    Science.gov (United States)

    Isupova, Olga G

    2011-01-01

    The article is concerned with the life experiences of infertile women going through infertility treatment and their need for social and psychological support, which they try to find in their immediate social environment. The Internet has become one place where everyone can find "people like oneself." The best support is received from these people who are in the same life situation and are able and willing to share their lived experiences with each other. Communication via the Internet and the formation of a virtual community of patients has both positive and negative aspects, all of which are examined in the article. On the one hand, it creates a psychologically favorable atmosphere and might potentially increase the success rate of IVF treatment. On the other, this leads to the seclusion of patients within the circle of "similar people" and sometimes to negative attitudes towards people outside the circle. The article is based on the author's "netnography" research of a virtual community of Russian In-Vitro Fertilization (IVF)(1) patients.

  16. Anesthesia Related Toxic Effects on In Vitro Fertilization Outcome: Burden of Proof

    Directory of Open Access Journals (Sweden)

    Paraskevi Matsota

    2015-01-01

    Full Text Available Management of pain and anxiety during oocyte retrieval makes anesthesia an important part of the in vitro fertilization (IVF procedures. There are many studies investigating the influence of anesthesia on IVF success. This review article provides an overview of published data regarding the potential toxic effects of different anesthetic techniques (Loco-regional, general anesthesia (GA, and monitored anesthesia care (MAC, different anesthetic agents, and alternative medicine approach (principally acupuncture on the IVF outcome. From our analysis, evidence of serious toxicity in humans is not well established. Trials regarding different anesthetic techniques ended up without clear conclusions. Studies about GA came up with conflicting results. A few trials relate GA with lower pregnancy rates, although some others failed to prove this conclusion. Furthermore, detectable amounts of some anesthetic agents are measurable in the follicular fluid but these findings are not strongly associated with toxicity. MAC and Loco-regional anesthesia appear as safe alternative choices and there is evidence of improved outcome. Whereas acupuncture may provide assistance increasing IVF success according to some trials, some others could not obtain these effects. Questions about the appropriate time of application and the underlying mechanism of action are not answered yet, so further investigation should be done.

  17. In vitro fertilization using frozen-thawed feline epididymal spermatozoa from corpus and cauda regions.

    Science.gov (United States)

    Kunkitti, Panisara; Axnér, Eva; Bergqvist, Ann-Sofi; Sjunnesson, Ylva

    2016-10-01

    Epididymal sperm preservation offers a potential for rescuing genetic material from endangered or valuable animals after injury or death. Spermatozoa from corpus, as well as from cauda, have the capability to be motile and to undergo capacitation and can thus potentially be preserved for assisted reproductive technologies. In the present study, feline frozen-thawed epididymal spermatozoa from corpus and cauda regions were investigated for their ability to fertilize homologous oocytes and further embryo development in vitro. Epididymal spermatozoa from corpus and cauda of seven cats were cryopreserved and used for IVF. Cumulus-oocyte complexes (n = 419) were obtained from female cats after routine spaying. Frozen-thawed corpus epididymal spermatozoa showed similar properties of acrosome integrity, membrane integrity, and chromatin integrity as frozen-thawed spermatozoa from cauda except corpus spermatozoa showed lower motility (P epididymal spermatozoa was confirmed by similar number of embryos developing to the two- and four-cell stages compared with sperm from cauda (32.03% vs. 33.33%). However, oocytes fertilized with corpus spermatozoa had lower potential to develop to the blastocyst stage (6.79%) and had lower cell numbers compared to oocytes fertilized with cauda spermatozoa (14.08%). In conclusion, spermatozoa from corpus epididymis had a similar capability to fertilize homologous oocytes in vitro as sperm from cauda but resulted in fewer embryos developing to the blastocyst stage compared to spermatozoa from the cauda. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. The correlation between endometrial thickness and outcome of in vitro fertilization and embryo transfer (IVF-ET) outcome

    OpenAIRE

    Al-Rejjal Rafat; Al-Hassan Saad; Coskun Serdar; Al-Ghamdi Ahlam; Awartani Khalid

    2008-01-01

    Abstract Background To evaluate the relationship between endometrial thickness on day of human chorionic gonadotrophin administration (hCG) and pregnancy outcome in a large number of consecutive in vitro fertilization and embryo transfer (IVF-ET) cycles. Methods A retrospective cohort study including all patients who had IVF-ET from January 2003–December 2005 conducted at a tertiary center. Results A total of 2464 cycles were analysed. Pregnancy rate (PR) was 35.8%. PR increased linearly (r =...

  19. EVALUATION OF IN VITRO FERTILITY OF SEMEN FROM YOUNG LOCAL BULLS IN OOCYTES FROM OVARIES OF SLAUGHTERED ANIMALS

    OpenAIRE

    Cabrera V., Próspero; Departamento de Producción Animal, Facultad de Zootecnia, Universidad Nacional Agraria La Molina (UNALM), Lima; Yoong K., Washington; Departamento de Producción Animal, Universidad Nacional Agraria La Molina, Lima; Gamarra L., Gicell; Investigador del Centro de Investigación y Enseñanza en Transferencia de Embriones (CIETE), Lima

    2012-01-01

    This study was carried out in the Center of Research and Instruction in Embryo Transfer (CIETE), sponsored by a partnership between the National Agrarian University of La Molina (UNALM) and the Ministry of Agriculture (MINAG), located at the UNALM campus. The objective was to evaluate the fertility and in vitro production of embryos of four bulls from the National Semen Bank. This study used 1031 oocytes of quality A and B obtained from ovaries of the local slaughterhouse. The oocytes were ma...

  20. Poor Prognosis with In Vitro Fertilization in Indian Women Compared to Caucasian Women Despite Similar Embryo Quality

    OpenAIRE

    Shahine, Lora K.; Lamb, Julie D.; Lathi, Ruth B.; Milki, Amin A.; Langen, Elizabeth; Westphal, Lynn M.

    2009-01-01

    BACKGROUND: Disease prevalence and response to medical therapy may differ among patients of diverse ethnicities. Poor outcomes with in vitro fertilization (IVF) treatment have been previously shown in Indian women compared to Caucasian women, and some evidence suggests that poor embryo quality may be a cause for the discrepancy. In our center, only patients with the highest quality cleavage stage embryos are considered eligible for extending embryo culture to the blastocyst stage. We compared...

  1. Porcine embryonic stem cells

    DEFF Research Database (Denmark)

    Hall, Vanessa Jane

    2008-01-01

    The development of porcine embryonic stem cell lines (pESC) has received renewed interest given the advances being made in the production of immunocompatible transgenic pigs. However, difficulties are evident in the production of pESCs in-vitro. This may largely be attributable to differences...

  2. New insights into the mechanisms of fertilization: comparison of the fertilization steps, composition, and structure of the zona pellucida between horses and pigs.

    Science.gov (United States)

    Mugnier, Sylvie; Dell'Aquila, Maria Elena; Pelaez, Jesus; Douet, Cécile; Ambruosi, Barbara; De Santis, Teresa; Lacalandra, Giovanni Michele; Lebos, Claude; Sizaret, Pierre-Yves; Delaleu, Bernadette; Monget, Philippe; Mermillod, Pascal; Magistrini, Michèle; Meyers, Stuart A; Goudet, Ghylène

    2009-11-01

    The mechanism of fertilization remains largely enigmatic in mammals. Most studies exploring the molecular mechanism underlying fertilization have been restricted to a single species, generally the mouse, without a comparative approach. However, the identification of divergences between species could allow us to highlight key components in the mechanism of fertilization. In the pig, in vitro fertilization (IVF) and polyspermy rates are high, and spermatozoa penetrate easily through the zona pellucida (ZP). In contrast, IVF rates are low in the horse, and polyspermy is scarce. Our objective was to develop a comparative strategy between these two divergent models. First, we compared the role of equine and porcine gametes in the following five functions using intraspecific and interspecific IVF: ZP binding, acrosome reaction, penetration through the ZP, gamete fusion, and pronucleus formation. Under in vitro conditions, we showed that the ZP is a determining element in sperm-ZP attachment and penetration, whereas the capacity of the spermatozoa is of less importance. In contrast, the capacity of the spermatozoa is a key component of the acrosome reaction step. Second, we compared the composition and structure of the equine and porcine ZP. We observed differences in the number and localization of the ZP glycoproteins and in the mesh-like structure of the ZP between equine and porcine species. These differences might correlate with the differences in spermatozoal attachment and penetration rates. In conclusion, our comparative approach allows us to identify determining elements in the mechanism of fertilization.

  3. Effect of Phosphate Fertilizer and Arbuscular Mycorrhizal Fungi on The Nutrient, Phosphateuptake and in Vitro Digestibility of Alfalfa

    Directory of Open Access Journals (Sweden)

    Bambang Suwignyo

    2016-11-01

    Full Text Available This study aimed to determine the effect of arbuscular mycorrhizal fungi (AMF and phosphate (P fertilizer on the nutrient content, phosphate uptake and in vitro digestibility of alfalfa (Medicago sativa L..The research was conducted at green house of Forage and Pastures Science Laboratory, Faculty of Animal Science Universitas Gadjah Mada. The experiment was arranged in Completely Randomized Design using 3x4 factorial patterns with four replications. The first factor was dosage of phosphate fertilizer SP 36 (0, 60, and 120 kg/ha. Second factor was the dosage of AMF (0, 0.8, 1.6, and 2.4kg/ha. The variable measured was nutrient contents (crude protein, dry matter, and organic matter, total P uptake and dry matter and organic matter in vitro digestibility. The results showed that the interaction of AMF and P fertilizer had no significant effect on crude protein and total P uptake, but highly significant effect on the parameters of dry matter, organic matter and dry matter and organic matter in vitro digestibility.

  4. Psychological correlates of prenatal attachment in women who conceived after in vitro fertilization and women who conceived naturally.

    Science.gov (United States)

    Hjelmstedt, Anna; Widström, Ann-Marie; Collins, Aila

    2006-12-01

    Investigators have pointed out that long-awaited pregnancies, such as those after in vitro fertilization (IVF), are emotionally vulnerable. In addition, higher pregnancy-related distress has been found among women pregnant after in vitro fertilization compared with women with "naturally" achieved pregnancy. The aim of this study was to compare prenatal attachment among IVF mothers and control mothers (women who conceived naturally), and to study relationships between prenatal attachment and psychosocial variables. Fifty-six IVF women from IVF clinics and 41 control women from antenatal clinics in Stockholm were assessed in gestational weeks 26 and 36. They completed self-rating scales measuring prenatal attachment, personality, marital relationship, anxiety, and depression. Prenatal attachment increased as the pregnancy progressed in both groups. Prenatal attachment rated in gestational week 26 was significantly associated with that in gestational week 36. Multiple regression analyses showed that, in gestational week 26, prenatal attachment was explained by satisfaction with the partner relationship, whereas in gestational week 36 the factors contributing to high prenatal attachment were low scores of the personality trait detachment, low ambivalence, and younger age. Method of conception was unrelated to prenatal attachment at either assessment time point. In vitro fertilization mothers are attached to their unborn children to the same extent as other mothers. Prenatal attachment increases during pregnancy. At the same time, however, individual scores on prenatal attachment seem to be relatively stable. Significant contributors to prenatal attachment are marital satisfaction, age, ambivalence, and detachment.

  5. Preterm delivery risk factors in singletons born after in vitro fertilization procedures.

    Science.gov (United States)

    Ban Frangez, Helena; Korosec, Sara; Verdenik, Ivan; Kotar, Vanja; Kladnik, Urska; Vrtacnik Bokal, Eda

    2014-05-01

    Women delivering singletons after in vitro fertilization (IVF) procedures have a greater risk of preterm delivery (PD). The aim of our study was to analyze PD risk factors and to identify those that could possibly be prevented. In our matched controlled study we analyzed 1127 singleton deliveries after IVF and transfer of fresh embryos performed at the University Medical Center Ljubljana between 1 January 2002 and 31 December 2010. For every delivery included in the study group we chose three consecutive controls matched by maternal age, parity and maternity hospital. The main outcome measure was PD (anomaly, operation on the uterus, chronic renal disease, maternal age and parity, and body mass index (BMI). Variables investigated within the IVF group were: stimulation protocol, laboratory procedure, number of retrieved oocytes and number and quality of transferred embryos. The PD rate after IVF was 1.5 times higher than after natural conception (11.5% in the IVF group and 7.7% in the control group, p30 was an important risk factor only in the IVF group (OR 1.86 (1.06-3.27) vs. 1.10 (0.67-1.80)) and PPD only in the controls (OR 1.83 (0.78-4.28) vs. 3.22 (1.55-6.67)). Among the investigated PD risk factors, an IVF procedure was shown to be the fifth most important one. On analyzing parameters of the ovarian stimulation and IVF procedure, no PD risk factor was identified. IVF was shown to be a significant risk factor for PD. In the IVF population, BMI plays a far more important role in PD than in the fertile population. In our research PD reoccurrence in IVF group was less than expected, which could perhaps be explained by the surgical correction of gynecological pathology and, where necessary, its being combined with cerclage. The investigation of parameters related to the IVF procedure did not identify any risk factors for PD. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  6. Uncertainty in clinical data and stochastic model for in vitro fertilization.

    Science.gov (United States)

    Yenkie, Kirti M; Diwekar, Urmila

    2015-02-21

    In vitro fertilization (IVF) is the most widely used technique in assisted reproductive technologies (ART). It has been divided into four stages; (i) superovulation, (ii) egg retrieval, (iii) insemination/fertilization and (iv) embryo transfer. The first stage of superovulation is a drug induced method to enable multiple ovulation, i.e., multiple follicle growth to oocytes or matured follicles in a single menstrual cycle. IVF being a medical procedure that aims at manipulating the biological functions in the human body is subjected to inherent sources of uncertainty and variability. Also, the interplay of hormones with the natural functioning of the ovaries to stimulate multiple ovulation as against single ovulation in a normal menstrual cycle makes the procedure dependent on several factors like the patient's condition in terms of cause of infertility, actual ovarian function, responsiveness to the medication, etc. The treatment requires continuous monitoring and testing and this can give rise to errors in observations and reports. These uncertainties are present in the form of measurement noise in the clinical data. Thus, it becomes essential to look at the process noise and account for it to build better representative models for follicle growth. The purpose of this work is to come up with a robust model which can project the superovulation cycle outcome based on the hormonal doses and patient response in a better way in presence of uncertainty. The stochastic model results in better projection of the cycle outcomes for the patients where the deterministic model has some deviations from the clinical observations and the growth term value is not within the range of '0.3-0.6'. It was found that the prediction accuracy was enhanced by more than 70% for two patients by using the stochastic model projections. Also, in patients where the prediction accuracy did not increase significantly, a better match with the trend of the clinical data was observed in case of the

  7. The impact of bariatric surgery on obesity-related infertility and in vitro fertilization outcomes.

    Science.gov (United States)

    Tan, Orkun; Carr, Bruce R

    2012-12-01

    Obesity-related infertility is one of the most common problems of reproductive-age obese women who desire childbearing. The various types of bariatric surgeries have proved effective in controlling excessive weight gain, improving fertility, and preventing certain maternal and fetal complications in these women. This article summarizes the current evidence regarding the impact of bariatric surgery on obesity-related infertility and in vitro fertilization (IVF) outcomes. We have also attempted to draw conclusions about maternal and fetal risks and the benefits of bariatric surgery. Laparoscopic adjustable gastric banding and Roux-en-Y procedures are the two most commonly performed bariatric surgeries. Bariatric surgery was believed to improve menstrual irregularity and increase ovulation rate in anovulatory obese women, which lead to increased pregnancy rates. Although there are data in the literature suggesting the improvement of both the ovulatory function and the spontaneous pregnancy rates in obese women who lost weight after bariatric surgery, most of these are case-control studies with a small number of patients. The data are insufficient to determine an ideal time interval for pregnancy after bariatric surgery; however, the general consensus is that pregnancy should be delayed 12 to 18 months after bariatric surgery to avoid nutritional deficiencies. Few data exist regarding IVF success rates in women who have undergone bariatric surgery. One pairwise study discussed five patients who underwent bariatric surgery followed by IVF that resulted in three term pregnancies in three patients after the first IVF cycle. Many studies reported reductions in obesity-related pregnancy complications such as gestational diabetes and hypertensive disorders after bariatric surgery. Although data are inconsistent, some studies reported increased rate of preterm delivery and small for gestational age infants after bariatric surgery. Pregnancies after bariatric surgery may be

  8. In vitro development of preimplantation porcine embryos using alginate hydrogels as a three-dimensional extracellular matrix

    Science.gov (United States)

    Between day 10 and 12 of gestation, porcine embryos undergo a dramatic morphological change, known as elongation, with a corresponding increase in estrogen production for maternal recognition of pregnancy. Elongation deficiencies contribute to ~20% of embryonic loss, but exact mechanisms of elongati...

  9. Fertility outcome of laparoscopic treatment in patients with severe endometriosis and repeated in vitro fertilization failures.

    Science.gov (United States)

    Soriano, David; Adler, Iris; Bouaziz, Jerome; Zolti, Matti; Eisenberg, Vered H; Goldenberg, Mordechai; Seidman, Daniel S; Elizur, Shai E

    2016-10-01

    To evaluate fertility outcomes in infertile women with severe endometriosis (The revised American Fertility Society classification [AFS] 3-4) and repeated IVF failures, who underwent surgery due to exacerbation of endometriosis-related symptoms. Retrospective cohort study. University hospital. All women who failed IVF treatment before surgery and who underwent laparoscopic surgery for severe endometriosis between January 2006 and December 2014. All patients were operated by highly skilled surgeons specializing in laparoscopic surgery for advanced endometriosis. Only patients with evidence of endometriosis in the pathology specimens were included in this study. Delivery rate after surgery. Seventy-eight women were included in the present study. All women were diagnosed with severe endometriosis during surgery (AFS 3-4) and all women had experienced failed IVF treatments before surgery. All women were symptomatic before their surgery. After surgical treatment 33 women (42.3%) delivered. Three women (9%) conceived spontaneously and all other women conceived after IVF treatment. Women who delivered were younger (32.5 [±4.1] years vs. 35.5 [±3.8] years), were less often diagnosed with diminished ovarian reserve before surgery (6% vs. 28.8%), and were more often diagnosed with normal uterine anatomy (by preoperative transvaginal ultrasound and during operation). In addition, performing salpingectomy during surgery was associated with a trend of improvement in delivery rates after surgery (70% in women who delivered vs. 51% in women who failed to deliver). Symptomatic women with severe endometriosis and repeated IVF implantation failures may benefit from extensive laparoscopic surgery when performed by an experienced multidisciplinary surgical team to improve IVF outcome. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  10. When isolated at full receptivity, in vitro fertilized wheat (Triticum aestivum, L.) egg cells reveal [Ca2+]cyt oscillation of intracellular origin

    National Research Council Canada - National Science Library

    Pónya, Zsolt; Corsi, Ilaria; Hoffmann, Richárd; Kovács, Melinda; Dobosy, Anikó; Kovács, Attila Zoltán; Cresti, Mauro; Barnabás, Beáta

    2014-01-01

    During in vitro fertilization of wheat (Triticum aestivum, L.) in egg cells isolated at various developmental stages, changes in cytosolic free calcium ([Ca2+]cyt) were observed. The dynamics of [Ca2...

  11. Effect of caffeine on motility and vitality of sperm and in vitro fertilization of outbreed mouse in T6 and M16 media

    National Research Council Canada - National Science Library

    Nabavi, Narges; Todehdehghan, Fatemeh; Shiravi, Abdollhossein

    2013-01-01

    Caffeine increases the CAMP production that stimulates spermatozoa movement. Caffeine is also used for induction of in vitro acrosome reaction in mammalian spermatozoa, an important step in achieving fertilization...

  12. Metabolomic profiling of human follicular fluid from patients with repeated failure of in vitro fertilization using gas chromatography/mass spectrometry

    National Research Council Canada - National Science Library

    Xia, Lan; Zhao, Xiaoming; Sun, Yun; Hong, Yan; Gao, Yuping; Hu, Shuanggang

    2014-01-01

    ...) from patients with in vitro fertilization (IVF) and repeated IVF failure (RIF). A prospective study was employed in Center for Reprodutive Medcine, Renji Hospital, Shanghai, China, between January and October 2010...

  13. Live-Birth Rate Associated With Repeat In Vitro Fertilization Treatment Cycles.

    Science.gov (United States)

    Smith, Andrew D A C; Tilling, Kate; Nelson, Scott M; Lawlor, Debbie A

    The likelihood of achieving a live birth with repeat in vitro fertilization (IVF) is unclear, yet treatment is commonly limited to 3 or 4 embryo transfers. To determine the live-birth rate per initiated ovarian stimulation IVF cycle and with repeated cycles. Prospective study of 156,947 UK women who received 257,398 IVF ovarian stimulation cycles between 2003 and 2010 and were followed up until June 2012. In vitro fertilization, with a cycle defined as an episode of ovarian stimulation and all subsequent separate fresh and frozen embryo transfers. Live-birth rate per IVF cycle and the cumulative live-birth rates across all cycles in all women and by age and treatment type. Optimal, prognosis-adjusted, and conservative cumulative live-birth rates were estimated, reflecting 0%, 30%, and 100%, respectively, of women who discontinued due to poor prognosis and having a live-birth rate of 0 had they continued. Among the 156,947 women, the median age at start of treatment was 35 years (interquartile range, 32-38; range, 18-55), and the median duration of infertility for all 257,398 cycles was 4 years (interquartile range, 2-6; range, live-birth rate for the first cycle was 29.5% (95% CI, 29.3%-29.7%). This remained above 20% up to and including the fourth cycle. The cumulative prognosis-adjusted live-birth rate across all cycles continued to increase up to the ninth cycle, with 65.3% (95% CI, 64.8%-65.8%) of women achieving a live birth by the sixth cycle. In women younger than 40 years using their own oocytes, the live-birth rate for the first cycle was 32.3% (95% CI, 32.0%-32.5%) and remained above 20% up to and including the fourth cycle. Six cycles achieved a cumulative prognosis-adjusted live-birth rate of 68.4% (95% CI, 67.8%-68.9%). For women aged 40 to 42 years, the live-birth rate for the first cycle was 12.3% (95% CI, 11.8%-12.8%), with 6 cycles achieving a cumulative prognosis-adjusted live-birth rate of 31.5% (95% CI, 29.7%-33.3%). For women older than 42

  14. Women's alcohol consumption and cumulative incidence of live birth following in vitro fertilization.

    Science.gov (United States)

    Dodge, L E; Missmer, S A; Thornton, K L; Hacker, M R

    2017-07-01

    The objective of this study was to determine the effect of alcohol consumption on outcomes among women undergoing in vitro fertilization (IVF). This study is a retrospective cohort study. This study was performed in a private academically affiliated IVF center. Patients included women presenting for their first IVF cycle from July 2004 through October 2012. Women completed self-administered questionnaires before their first IVF cycle, which included report of usual alcohol consumption. Women were categorized as non-drinkers, social drinkers, or daily drinkers, as well as by the number of drinks consumed per week. Competing risks analysis was used to calculate the cumulative incidence of live birth after 6 cycles stratified by alcohol consumption. Main outcome measures included spontaneous abortion, clinical pregnancy, and live birth following IVF. There were 591 (27.7%) non-drinkers, 1466 (68.7%) social drinkers, and 77 (3.6%) daily drinkers (total n = 2134). In the first cycle, compared to non-drinkers, daily drinkers had a twofold increased risk of spontaneous abortion (adjusted risk ratio [aRR] 2.2; 95% confidence interval [CI] 1.1-4.5) among all cycle starts, and while their risk of live birth was 30% lower (aRR 0.7; 95% CI 0.4-1.3), the sample size was small, and it was not significantly lower. By the end of 6 cycles, social drinkers and daily drinkers did not differ from non-drinkers in their cumulative incidence of live birth (56.1, 50.6, and 52.1%, respectively; both P ≥ 0.28). There was a trend towards lower risk of live birth among daily drinkers. Daily drinkers had an increased risk of spontaneous abortion in the first cycle, but the number of daily drinkers was small.

  15. Risk factors and predicting health disorders in infants born from monocyesis after in vitro fertilization

    Directory of Open Access Journals (Sweden)

    L.A. Pykhtina

    2017-03-01

    Full Text Available Issues of health state and factors determining it in infants born due to in vitro fertilization (IVF have been frequently discussed over recent years. The question yet to be answered is whether health disorders are related to burdened pre-morbid state of a mother as per extragenital and obstetric-gynecologic pathology or they are caused by IVF. Selective transfer of only one embryo has tended to increase recently, and, given this trend, specialists all around the world, and in our country as well, face a problem of detecting risk factors which cause health disorders in children born in monocyesis after IVF. Prognostic algorithm for most frequent pathologies is being worked out and it will help to implement targeted and differentiated approach to their prevention. We have completed clinical examination of infants during the first year of their life; all these infants were born in monocyesis after IVF (n=121. We have also questioned and interviewed their parents and analyzed data taken from infants' development records over a year as well. We have applied Wald sequential mathematical analysis to determine risk factors and create prognostic tables comprising most frequent somatic health disorders such as congenital abnormalities, iron-deficiency anemia, atopic dermatitis, absence of perinatal CNS damage compensation. We have detected that factors related to a mother's and newborn's health (extragenital morbidity, obstetric-gynecologic case history, pregnancy course exert their influence on health disorders evolvement in such children during their first year of life; we have also shown that social factors and factors associated with IVF procedure don't have any statistical significance.

  16. Reproductive Outcomes of Segmented In Vitro Fertilization in Patients Diagnosed with Endometriomas.

    Science.gov (United States)

    Ozgur, Kemal; Bulut, Hasan; Berkkanoglu, Murat; Coetzee, Kevin

    2018-01-01

    To assess the impact of ovarian endometriomas on endometrial receptivity in frozen embryo transfer (FET) of segmented in vitro fertilization (IVF) cycles. Retrospective, matched-control study (Canadian Task Force classification II-2). A single, private assisted-reproduction technology center. Thirty patients diagnosed with unilateral or bilateral endometriomas were compared with 60 patients without endometriomas in a population of 1894 patients who underwent segmented IVF treatment between September 2014 and September 2016. Intracytoplasmic sperm injection with blastocyst freeze-all and FET. The primary endpoint of the study was a viable pregnancy (>14 weeks). The mean diameter of diagnosed endometriomas was 25.7 ± 10.6 mm. The median antral follicle count was significantly lower in the endometrioma group compared with the entire study population (11.5; interquartile range [IQR], 6.0-17.0 vs 14.0; IQR, 9.0-22.0; p = .042). The median number of mature ovarian follicles (≥14 mm) per antral follicle that developed during controlled ovarian stimulation was not significantly different between the groups (11.0 [IQR, 5.8-14.3] vs 10.0 [IQR, 6.0-15.8]; p = .908); however, the median number of oocytes retrieved was lower in the endometrioma group (11.5 [IQR, 6.0-21.5] vs 13.5 [IQR, 9.0-20.8]; p = .373). The biochemical pregnancy, implantation, and ongoing pregnancy rates were not significantly different between the endometrioma and control groups. Although ovarian endometriomas result in reduced ovarian reserve and oocyte retrieval, their impact on reproductive outcome is limited with FET. Copyright © 2017 American Association of Gynecologic Laparoscopists. Published by Elsevier Inc. All rights reserved.

  17. Misdiagnosis and delayed diagnosis for ectopic and heterotopic pregnancies after in vitro fertilization and embryo transfer.

    Science.gov (United States)

    Wang, Lin-lin; Chen, Xin; Ye, De-sheng; Liu, Yu-dong; He, Yu-xia; Guo, Wei; Chen, Shi-ling

    2014-02-01

    This study examined the misdiagnosis and delayed diagnosis factors for ectopic pregnancy (EP) and heterotopic pregnancy (HP) after in vitro fertilization and embryo transfer (IVF-ET) in an attempt to reduce the diagnostic error. Clinical data of patients who underwent IVF-ET treatment and had clinical pregnancy from 12463 cycles were retrospectively analyzed. Their findings of serum β-hCG test and transvaginal ultrasonography were also obtained during follow-up. These patients were divided into two groups according to the diagnosis accuracy of EP/HP: early diagnosis and misdiagnosis/delayed diagnosis. The results showed that the incidence of EP and HP was 3.8% (125/3286) and 0.8% (27/3286) respectively for IVF/ICSI-ET cycle, and 3.8% (55/1431) and 0.7% (10/1431) respectively for frozen- thawed embryo transfer (FET) cycle. Ruptured EP occurred in 28 patients due to initial misdiagnosis or delayed diagnosis. Related factors fell in 3 categories: (1) clinician factors: misunderstanding of patients' medical history, insufficient training in ultrasonography and unawareness of EP and HP; (2) patient factors: noncompliance with medical orders and lack of communication with clinicians; (3) complicated conditions of EP: atypical symptoms, delayed elevation of serum β-hCG level, early rupture of cornual EP, asymptomatic in early gestation and pregnancy of unknown location. All the factors were interwoven, contributing to the occurrence of EP and HP. It was concluded that complicated conditions are more likely to affect the diagnosis accuracy of EP/HP after IVF-ET. Transvaginal ultrasonography should be performed at 5 weeks of gestation. Intensive follow-up including repeated ultrasonography and serial serum β-hCG tests should be performed in patients with a suspicious diagnosis at admission.

  18. In Vitro Fertilization Is Successful in Women With Ulcerative Colitis and Ileal Pouch Anal Anastomosis.

    Science.gov (United States)

    Pabby, Vikas; Oza, Sveta Shah; Dodge, Laura E; Hacker, Michele R; Moragianni, Vasiliki A; Correia, Katherine; Missmer, Stacey A; Fox, Janis H; Ibrahim, Yetunde; Penzias, Alan; Burakoff, Robert; Cheifetz, Adam; Friedman, Sonia

    2015-06-01

    Women with ulcerative colitis (UC), who require ileal pouch anal anastomosis (IPAA), have up to a threefold increased incidence of infertility. To better counsel patients who require colectomy, we examined the success rates of in vitro fertilization (IVF) among women who have undergone IPAA. This was a retrospective cohort study conducted at the Brigham and Women's Hospital and Beth Israel Deaconess Medical Center. Female patients with UC were identified via ICD-9 codes and cross-referenced with those presenting for IVF from 1998 through 2011. UC patients with IPAA were compared with the following two unexposed groups that underwent IVF: (1) patients with UC, who had not undergone IPAA, and (2) patients without inflammatory bowel disease (IBD). The primary outcome was the cumulative live birth rate. Secondary outcomes included number of oocytes retrieved, proportion of patients who underwent embryo transfer, pregnancy rate, and live birth rate at first cycle. There were 22 patients with UC and IPAA, 49 patients with UC and without IPAA, and 470 patients without IBD. The cumulative live birth rate after six cycles in the UC and IPAA groups was 64% (95% confidence interval (CI): 44-83%). This rate did not differ from the cumulative live birth rate in the UC without IPAA group (71%, 95% CI: 59-83%; P=0.63) or the group without IBD (53%, 95% CI: 48-57%; P=0.57). This study demonstrates that in our cohort, women who undergo IPAA achieve live births following IVF at comparable rates to women with UC without IPAA and to women without IBD.

  19. Oocyte donation in low responders to conventional ovarian stimulation for in vitro fertilization.

    Science.gov (United States)

    Remohi, J; Vidal, A; Pellicer, A

    1993-06-01

    To analyze endometrial response (endometrial dating and implantation) to exogenous administration of E2-valerate and P in women with low response to gonadotropins undergoing oocyte donation. Prospective study. A cycle in which endometrial specimens were obtained and subsequent cycles with ET were evaluated. The control group was made up of patients with premature ovarian failure (POF) undergoing the same procedure. In Vitro Fertilization program at the Instituto Valenciano de Infertilidad. A total of 37 women with low response to gonadotropins in previous cycles and 33 women with POF. First artificial cycle with E2-valerate and P in the absence of previous pituitary suppression to determine endometrial adequacy. Successive artificial cycles in which ET was performed on cycle day 17. Oocytes donated from infertile patients undergoing IVF. Serum steroid levels were measured during the artificial cycle. Histologic dating of the endometrium on cycle days 15 and 26. Ultrasonographically documented IVF-ET pregnancies. Postovulatory changes on cycle day 15 were observed in 36.4% of low responders treated with E2-valerate and P in the absence of simultaneous pituitary suppression. Pregnancy rates were higher in women with previous sufficiently (77.8%) or insufficiently (80%) estrogen-primed endometrium than in the cases showing postovulatory changes (37.5%). Pregnancy rates (PRs) per transfer were significantly higher in low responders (63.8%) than in patients with POF (37.2%). Patients with endometriosis had a 71.4% PR per transfer. Embryos derived from oocytes from polycystic ovaries had a 48.3% PR. Oocyte donation is a reliable alternative for women with low response to gonadotropins, including those with severe endometriosis. The efficacy of the steroid replacement regimen in controlling ovarian function may influence outcome. Thus, women with functional ovaries despite exogenous steroid replacement might be differently treated. Women with polycystic ovaries are an

  20. Impact of The Endometrioma on Ovarian Response and Pregnancy Rate in In Vitro Fertilization Cycles

    Directory of Open Access Journals (Sweden)

    Mahnaz Ashrafi

    2014-03-01

    Full Text Available Background: Our objective was to evaluate the effect of ovarian endometrioma on ovarian stimulation outcomes in in vitro fertilization cycles (IVF. Materials and Methods: In this prospective cohort study, we followed 103 patients who underwent intra-cytoplasmic sperm injection (ICSI procedures over a 24-months period. The study group consisted of 47 infertile women with either unilateral or bilateral ovarian endometrial cysts of less than 3 cm. The control group consisting of 57 patients with mild male factor infertility was candidate for ICSI treatment during the same time period as the study groups. Both groups were compared for number of oocytes retrieved, grades of oocytes, as well as embryo quantity and quality. Results: Our results showed similar follicle numbers, good embryo grades (A or B and pregnancy rates in the compared groups. However, patients with endometrioma had higher gonadotropin consumption than the control group. The mean number of retrieved oocytes in patients with endometrioma was significantly lower than control group (6.6 ± 3.74 vs. 10.4 ± 5.25 (p<0.001. In addition, patients with endometrioma had significantly lower numbers of metaphase II (MII oocytes (5 ± 3.21 than controls (8.2 ± 5.4 (p<0.001. In patients with unilateral endometrioma, there were no significant differences in main outcome measures between normal and involved ovaries in the patients with endometrioma. Conclusion: Patients with ovarian endometrioma had poor outcome. They showed poor ovarian response with lower total numbers of retrieved oocytes and lower MII oocytes during the stimulation phase; however, it does not affect the total number of embryos transferred per patient, quality of embryos, and pregnancy rate per patient.

  1. Parenting style of women who conceived using in vitro fertilization: a meta-analysis.

    Science.gov (United States)

    Wang, Yu-Ming; Shu, Bih-Ching; Fetzer, Susan; Chang, Ying-Ju

    2014-06-01

    Research has shown that the mental health of women contributes to their parenting style. However, it remains unclear whether the experience of in vitro fertilization (IVF) affects parenting style. This study was designed to assess whether there is a difference in parenting styles between women who conceived using IVF and those who conceived naturally. This meta-analysis searched three electronic databases (MEDLINE, PsychInfo, and CINAHL) for relevant articles published between 1978 and 2011. Key words used included parenting, mothering, parent-child relations, childrearing, infertility, assisted reproductive technique, IVF, and intracytoplasmic sperm injection. Study inclusion criteria were as follows: published in an English-language peer-reviewed journal, with the definition of parenting style categorized as one of two dimensions: warmth and control; quantification of the parenting behaviors; use of a case-controlled study design to compare IVF and natural conceptions; and reported data sufficient to calculate the effect sizes. Studies that included women who conceived using a donor egg or sperm for IVF and those that included women who were either surrogates or in homosexual relationships were excluded. Three hundred ninety studies were identified. Fourteen studies met the inclusion criteria. The Newcastle-Ottawa Scale was used to appraise the quality of the data. The IVF participants used significantly greater controlling parenting behaviors than their natural conception participant peers (d = 0.148, p parenting behaviors related to warmth, rejection, or respect for autonomy. The homogeneity test for the effect size of warmth and controlling parenting behavior achieved significance. Women who conceive using IVF have slightly but still significantly greater controlling parenting behaviors than women who conceive naturally. The results of this study may help professionals to better understand the parenting style of IVF women and develop appropriate interventions

  2. Lessons from elective in vitro fertilization (IVF in, principally, non-infertile women

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    Gleicher Norbert

    2012-06-01

    Full Text Available Abstract Background We here report the first investigation of exclusively elective in vitro fertilization (IVF cycles in women with no apparent history of infertility. Since IVF outcome in women with infertility are always influenced by underlying causes of infertility, a study on non-infertile women may offer new insights. Methods We investigated 88 females without history of infertility in 109 consecutive elective IVF cycles, almost exclusively performed for purposes of preimplantation genetic screening (PGS; i.e., elective gender selection. The following questions were addressed: (i impact of PGS on IVF pregnancy chances; (ii impact of transfer of 1 vs. ≥2 embryos on IVF pregnancy chances; (iii correlation of anti-Müllerian hormone (AMH levels to embryo ploidy (iv effect of gonadotropin dosage used in stimulation on available embryos for transfer; and (v in form of a 1:1 case control study, compared 33 elective PGS cycles with matched control cycles without PGS, performed in couples with either prior tubal ligations and/or severe male factor infertility as indication of IVF. Results The overall clinical pregnancy rate for the group was 36.7%; pregnancy was associated with number of euploid (P = 0.009 and number of embryos transferred (P = 0.001. Odds of pregnancy were 3.4-times higher if ≥4 euploid embryos were produced in comparison to Conclusions This study suggests that outcomes of elective IVF cycles may significantly deviate from infertility-associated cycles. Affirming proof of concept for PGS, utilizing day-3 embryo biopsy and fluorescence in-situ hybridization (FISH, both widely held responsible for earlier failures to establish such proof, suggests that the principal cause of prior failures were likely not insufficient laboratory techniques but poor patient selection for PGS. Such a conclusion questions the current reintroduction of PGS with improved techniques and technologies in absence of prior determination of suited

  3. Does the Ovarian Stimulation Phase Length Predict In vitro Fertilization Outcomes?

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    Angela Baerwald

    2011-01-01

    Full Text Available Background: Bi-directional communication between the follicle and oocyte is necessary toregulate follicle and oocyte development. Currently, it is not practical to monitor the serial growthof individual follicles during assisted reproduction. The ovarian stimulation phase length (SPLis an indirect measure of mean follicular growth rate. The objective of this study was to test thehypothesis that a short or long SPL would be associated with suboptimal outcomes in womenundergoing in vitro fertilization (IVF.Materials and Methods: A retrospective cohort study was conducted in 140 women who underwentIVF. Follicle development was monitored every 2-3 days during ovarian stimulation usingtransvaginal ultrasonography. Once > 3 follicles reached ≥ 17 mm, human chorionic gonadotropin(hCG was administered. Oocyte retrieval was performed approximately 35 hours after hCG.Oocytes underwent IVF on the day of collection and were evaluated daily thereafter. Embryos weretransferred on days 3 or 5, depending on the number and quality of embryos available. Associationsbetween SPL, age, follicle, oocyte, embryo and pregnancy outcomes were evaluated (SPSS version17.0; SPSS Inc., Chicago, IL, USA.Results: A SPL of 11 days was associated with an optimal number of follicles that developed to≥ 6 mm, ≥ 10 mm and ≥ 15 mm; serum estradiol concentrations; and number of oocytes collected(p0.05. Associations betweenSPL and outcomes were not influenced by age (p>0.05.Conclusion: The ovarian SPL can be used to predict the number of follicles that develop, oocytescollected and serum estradiol concentrations, but not embryo or pregnancy outcomes.

  4. Predicting Implantation Outcome of In Vitro Fertilization and Intracytoplasmic Sperm Injection Using Data Mining Techniques.

    Science.gov (United States)

    Hafiz, Pegah; Nematollahi, Mohtaram; Boostani, Reza; Namavar Jahromi, Bahia

    2017-10-01

    In vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) are two important subsets of the assisted reproductive techniques, used for the treatment of infertility. Predicting implantation outcome of IVF/ICSI or the chance of pregnancy is essential for infertile couples, since these treatments are complex and expensive with a low probability of conception. In this cross-sectional study, the data of 486 patients were collected using census method. The IVF/ICSI dataset contains 29 variables along with an identifier for each patient that is either negative or positive. Mean accuracy and mean area under the receiver operating characteristic (ROC) curve are calculated for the classifiers. Sensitivity, specificity, positive and negative predictive values, and likelihood ratios of classifiers are employed as indicators of performance. The state-of-art classifiers which are candidates for this study include support vector machines, recursive partitioning (RPART), random forest (RF), adaptive boosting, and one-nearest neighbor. RF and RPART outperform the other comparable methods. The results revealed the areas under the ROC curve (AUC) as 84.23 and 82.05%, respectively. The importance of IVF/ICSI features was extracted from the output of RPART. Our findings demonstrate that the probability of pregnancy is low for women aged above 38. Classifiers RF and RPART are better at predicting IVF/ICSI cases compared to other decision makers that were tested in our study. Elicited decision rules of RPART determine useful predictive features of IVF/ICSI. Out of 20 factors, the age of woman, number of developed embryos, and serum estradiol level on the day of human chorionic gonadotropin administration are the three best features for such prediction.

  5. Effect of Somatic Cell Types and Culture Medium on in vitro Maturation, Fertilization and Early Development Capability of Buffalo Oocytes

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    H. Jamil*, H. A. Samad, N. Rehman, Z. I. Qureshi and L. A. Lodhi

    2011-04-01

    Full Text Available This study was designed to evaluate the efficacy of different somatic cell types and media in supporting in vitro maturation (IVM, in vitro fertilization (IVF and early embryonic development competence of buffalo follicular oocytes. Cumulus oocyte complexes were collected for maturation from follicles (>6mm of buffalo ovaries collected at the local abattoir. Oocytes were co-cultured in tissue culture medium (TCM-199 with either granulosa cells, cumulus cells, or buffalo oviductal epithelial cells (BOEC @ 3x106 cells/ml or in TCM-199 without helper cells (control at 39°C and 5%CO2 in humidified air. Fresh semen was prepared in modified Ca++ free Tyrode medium. Fertilization was carried out in four types of media: i Tyrode lactate albumin pyruvate (TALP, ii TALP+BOEC, iii modified Ca++ free Tyrode and iv modified Ca++ free Tyrode+BOEC. Fertilized oocytes were cultured for early embryonic development in TCM-199 with and without BOEC. Higher maturation rates were observed in the granulosa (84.24% and cumulus cells (83.44% than BOEC co culture system (73.37%. Highest fertilization rate was obtained in modified Ca++ free Tyrode with BOEC co culture (70.42%, followed by modified Ca++ free Tyrode alone (63.77%, TALP with BOEC (36.92% and TALP alone (10.94%. Development of early embryos (8-cell stage improved in TCM-199 with BOEC co culture than TCM-199 alone. From the results of this study, it can be concluded that addition of somatic cells (granulosa cells, cumulus cells results in higher maturation rates of buffalo follicular oocytes than BOEC co culture system, while fertilization rate improved in modified Ca++ free Tyrode with and without BOEC. Addition of BOEC to TCM-199 improved the developmental capacity of early embryo.

  6. In vitro, ex vivo and in vivo examination of buccal absorption of metoprolol with varying pH in TR146 cell culture, porcine buccal mucosa and Göttingen minipigs

    DEFF Research Database (Denmark)

    Holm, René; Meng-Lund, Emil; Andersen, Morten B.

    2013-01-01

    This work studied the buccal absorption of metoprolol in vitro, ex vivo and in vivo as a function of buffered pH at 7.4, 8.5, 9.0 and 9.5. Permeability studies showed a correlation (r(2)=0.92) between in vitro TR146 cell culture and ex vivo porcine buccal mucosa in a modified Ussing chamber...

  7. Effects of a mindfulness-based intervention on fertility quality of life and pregnancy rates among women subjected to first in vitro fertilization treatment.

    Science.gov (United States)

    Li, Jing; Long, Ling; Liu, Yu; He, Wei; Li, Min

    2016-02-01

    Generally, undergoing an in vitro fertilization (IVF) treatment is an emotional and physical burden for the infertile woman, which may negatively influence the treatment outcome. We conducted a study to investigate the effectiveness of a mindfulness-based intervention (MBI) among women subjected to first IVF treatment at a fertility medical center in China. Among infertile women registered for their first IVF treatment, 58 completed the intervention, and 50 were assigned to a control group using a non-randomized controlled study. Standardized measures of mindfulness, self-compassion, emotion regulation difficulties, infertility-related coping strategies and fertility quality of life (FertiQoL) were endorsed pre- and post-MBI, and measure of pregnancy rates at the sixth months after the intervention. Both groups were shown to be equivalent at baseline. By the end of the intervention, women who attended the intervention revealed a significant increase in mindfulness, self-compassion, meaning-based coping strategies and all FertiQoL domains. Inversely, they presented a significant decrease in emotion regulation difficulties, active- and passive-avoidance coping strategies. Women in the control group did not present significant changes in any of the psychological measures. Moreover, there were statistically significant differences between participants in the pregnancy rates, the experiment group higher than the control group. Being fully aware of the present moment without the lens of judgment, seems to help women relate to their infertility and IVF treatment in new ways. This is beneficial for promoting their self-compassion, adaptive emotion regulation and infertility-related coping strategies, which, in turn, may influence the FertiQoL and pregnancy rates. The brief and nonpharmaceutical nature of this intervention makes it a promising candidate for women' use during first IVF treatment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. In vitro fertilization and sperm cryopreservation in the black-footed cat (Felis nigripes) and sand cat (Felis margarita).

    Science.gov (United States)

    Herrick, J R; Campbell, M; Levens, G; Moore, T; Benson, K; D'Agostino, J; West, G; Okeson, D M; Coke, R; Portacio, S C; Leiske, K; Kreider, C; Polumbo, P J; Swanson, W F

    2010-03-01

    Studies of in vitro fertilization (IVF) and sperm cryopreservation have been conducted in several small cat species, but virtually no data exist for black-footed cats (Felis nigripes) (BFCs) or sand cats (Felis margarita) (SCs). The objectives of this study were 1) to compare in vitro motility and acrosome status of fresh and cryopreserved (frozen in pellets on dry ice or in straws in liquid nitrogen vapor) BFC and SC spermatozoa cultured in feline-optimized culture medium (FOCM) or Ham F-10, 2) to assess ovarian responsiveness in BFCs and SCs following exogenous gonadotropin treatment and laparoscopic oocyte recovery, and 3) to evaluate the fertility of fresh and frozen-thawed spermatozoa from both species using homologous and heterologous (domestic cat oocytes) IVF in the two culture media. Motility and acrosomal integrity of fresh and frozen-thawed spermatozoa from BFCs and SCs were similar (P > 0.05) in both media during 6 h of culture. Although effects were more pronounced in SCs, cryopreservation in straws was superior (P 80% of recovered oocytes were of optimal (grade 1) quality. The BFC and SC spermatozoa fertilized 60.0%-79.4% of homologous and 37.7%-42.7% of heterologous oocytes in both culture media, with increased (P < 0.05) cleavage of homologous (SC) and heterologous (BFC and SC) oocytes in FOCM. These results provide the first information to date on the gamete biology of two imperiled cat species and further our capacity to apply reproductive technologies for their conservation.

  9. In Vitro Gender-Dependent Inhibition of Porcine Cytochrome P450 Activity by Selected Flavonoids and Phenolic Acids

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    Bo Ekstrand

    2015-01-01

    Full Text Available We investigated gender-related differences in the ability of selected flavonoids and phenolic compounds to modify porcine hepatic CYP450-dependent activity. Using pools of microsomes from male and female pigs, the inhibition of the CYP families 1A, 2A, 2E1, and 3A was determined. The specific CYP activities were measured in the presence of the following selected compounds: rutin, myricetin, quercetin, isorhamnetin, p-coumaric acid, gallic acid, and caffeic acid. We determined that myricetin and isorhamnetin competitively inhibited porcine CYP1A activity in the microsomes from both male and female pigs but did not affect the CYP2A and CYP2E1. Additionally, isorhamnetin competitively inhibited CYP3A in both genders. Noncompetitive inhibition of CYP3A activity by myricetin was observed only in the microsomes from male pigs, whereas CYP3A in female pigs was not affected. Quercetin competitively inhibited CYP2E1 and CYP1A activity in the microsomes from male pigs and irreversibly CY3A in female pigs. No effect of quercetin on CYP2E1 was observed in the microsomes from female pigs. Neither phenolic acids nor rutin affected CYP450 activities. Taken together, our results suggest that the flavonoids myricetin, isorhamnetin, and quercetin may affect the activities of porcine CYP1A, CYP3A, and CYP2E1 in a gender-dependent manner.

  10. Sperm characteristics and in vitro fertilization ability of thawed spermatozoa from Black Manchega ram: electroejaculation and postmortem collection.

    Science.gov (United States)

    García-Alvarez, O; Maroto-Morales, A; Martínez-Pastor, F; Garde, J J; Ramón, M; Fernández-Santos, M R; Esteso, M C; Pérez-Guzmán, M D; Soler, A J

    2009-07-15

    The aim of this study was to assess two models of sperm collection on the quality and fertility of thawed spermatozoa from Black Manchega rams, a threatened breed. Sperm samples were collected by electroejaculation and postmortem from each male. Samples were diluted with Biladyl and frozen. Motility (subjective and objective by means of computer-assisted semen analysis), membrane integrity, and acrosomal status (microscopy) were assessed on fresh and thawed semen; plasmalemma integrity, mitochondrial membrane potential, DNA integrity, and acrosomal status were evaluated by flow cytometry on thawed semen. Thawed spermatozoa were used in a heterologous in vitro fertilization test. After thawing, the proportion of live spermatozoa with intact membrane (YO-PRO-1-/PI-) was higher for postmortem samples (Pelectroejaculated samples (P=0.026 and P=0.003). Both electroejaculated and postmortem samples fertilized oocytes. Nevertheless, electroejaculated samples yielded a higher percentage of hybrid embryos (P=0.041). In conclusion, although postmortem spermatozoa had better sperm quality after thawing, electroejaculated spermatozoa showed higher ratios for sperm quality when only the live population was considered. Electroejaculated and postmortem samples might be used for germplasm banking of this threatened breed, but the fertility of postmortem spermatozoa might be lower.

  11. Microtubule assembly and in vitro development of bovine oocytes with increased intracellular glutathione level prior to vitrification and in vitro fertilization.

    Science.gov (United States)

    Hara, H; Yamane, I; Noto, I; Kagawa, N; Kuwayama, M; Hirabayashi, M; Hochi, S

    2014-11-01

    Although vitrification is a useful technique for preservation of bovine oocytes, the yield of blastocysts derived from the vitrified oocytes is still low. We have recently reported a new type of cryoinjury, multiple aster formation, by which pronuclear migration and development of vitrified-warmed and in vitro-fertilized bovine oocytes are impaired. The aim of the present study was to investigate the effect of glutathione (GSH) content of vitrified bovine oocytes on multiple aster formation and subsequent in vitro development. Treatment of bovine cumulus-oocyte complexes with β-mercaptoethanol (βME) and L-cysteine (Cys) during in vitro maturation resulted in 2.5-fold higher GSH content not only in fresh control but also in vitrified-warmed oocytes. The percentage of normally fertilized zygotes exhibiting sperm aster(s) was >95% in all four groups (with or without βME/Cys × fresh control or vitrified). The frequency of multiple aster formation in vitrified oocytes (three-fold higher than that in fresh control oocytes) was not affected by the increased level of intracellular GSH with βME/Cys. Consequently, the migration and development of pronuclei as well as the yield of blastocysts from vitrified-warmed oocytes (17 versus 41%) were not improved. In addition, there was no effect of increased GSH level on the yield of blastocysts in fresh control groups.

  12. Investigating the Role of Surface Materials and Three Dimensional Architecture on In Vitro Differentiation of Porcine Monocyte-Derived Dendritic Cells

    DEFF Research Database (Denmark)

    Hartmann, Sofie Bruun; Mohanty, Soumyaranjan; Skovgaard, Kerstin

    2016-01-01

    In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured....... In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs) was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three......-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene) differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material...

  13. In vitro culture and characterization of putative porcine embryonic germ cells derived from domestic breeds and yucatan mini pig embryos at days 20-24 of gestation

    DEFF Research Database (Denmark)

    Petkov, Stoyan Gueorguiev; Marks, Hendrik; Klein, Tino

    2011-01-01

    Embryonic germ cells (EGC) are cultured pluripotent cells derived from primordial germ cells (PGC). This study explored the possibility of establishing porcine EGC from domestic breeds and Yucatan mini pigs using embryos at Days 17-24 of gestation. In vitro culture of PGC from both pooled......-Seq expression profiling showed no expression of the core pluripotency markers OCT4, SOX2, and NANOG, although most other pluripotency genes were expressed at levels comparable to those of mouse embryonic stem cells (ESC). Moreover, germ-specific genes such as BLIMP1 retained their expression. Functional......, their injection into immunodeficient mice did not result in teratoma formation. Our results suggest that the PGC-derived cells described in this study are EGC-like, but seem to be multipotent rather than pluripotent cells. Nevertheless, the thorough characterization of these cells in this study, and especially...

  14. Effect of co-culture canine cumulus and oviduct cells with porcine oocytes during maturation and subsequent embryo development of parthenotes in vitro.

    Science.gov (United States)

    Lee, Seok Hee; Oh, Hyun Ju; Kim, Min Jung; Kim, Geon A; Choi, Yoo Bin; Jo, Young Kwang; Setyawan, Erif Maha Nugraha; Lee, Byeong Chun

    2018-01-15

    In the estrus stage, canine oocytes are surrounded by cumulus cells and undergo maturation in the oviduct for 2-3 days after ovulation. We hypothesized that canine oviduct cells (cOC) and canine cumulus cells (cCC) during this stage might affect the maturation of oocytes and thereby improve subsequent embryo development. Therefore, the objective of this study was to compare the effects of a cOC and cCC co-culture on oocyte in vitro maturation (IVM) and subsequent embryo development, and to analyze the gene expressions in a molecular fashion what co-culture actually gives the specific pathways in which the co-culture cells act to improve maturation and embryo development. The effect of co-culture using cOC and cCC on porcine oocyte IVM was investigated. Thereafter, oocytes were activated using electrical stimulation and embryo developmental competence was estimated. The expression of the genes related to oocyte maturation, embryo development and apoptosis were analyzed. Also, reactive oxygen species (ROS) levels after IVM was analyzed. The IVM rate and embryo development including cleavage, blastocyst formation rates, and total blastocyst cell numbers from cOC group were significantly higher than other groups (P culture with cOC improved in vitro porcine oocyte maturation and subsequent embryo development competence. Also, co-culture with cOC during IVM induces a suitable environment for oocyte maturation by enhancing the mRNA level of SMAD2/3 and GDF9, and for embryo development by elevating the expression level of PTGS2, WNT3A and MMP2. In addition, the decreased ROS level in cOC co-culture could have a beneficial influence on oocyte maturation. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. In vitro culture and characterization of putative porcine embryonic germ cells derived from domestic breeds and Yucatan mini pig embryos at Days 20-24 of gestation.

    Science.gov (United States)

    Petkov, Stoyan G; Marks, Hendrik; Klein, Tino; Garcia, Rodrigo S; Gao, Yu; Stunnenberg, Henk; Hyttel, Poul

    2011-05-01

    Embryonic germ cells (EGC) are cultured pluripotent cells derived from primordial germ cells (PGC). This study explored the possibility of establishing porcine EGC from domestic breeds and Yucatan mini pigs using embryos at Days 17-24 of gestation. In vitro culture of PGC from both pooled and individual embryos resulted in the successful derivation of putative EGC lines from Days 20 to 24 with high efficiency. RT-PCR showed that gene expression among all 31 obtained cell lines was very similar, and only minor changes were detected during in vitro passaging of the cells. Genome-wide RNA-Seq expression profiling showed no expression of the core pluripotency markers OCT4, SOX2, and NANOG, although most other pluripotency genes were expressed at levels comparable to those of mouse embryonic stem cells (ESC). Moreover, germ-specific genes such as BLIMP1 retained their expression. Functional annotation clustering of the gene expression pattern of the putative EGC suggests partial differentiation toward endo/mesodermal lineages. The putative EGC were able to form embryoid bodies in suspension culture and to differentiate into epithelial-like, mesenchymal-like, and neuronal-like cells. However, their injection into immunodeficient mice did not result in teratoma formation. Our results suggest that the PGC-derived cells described in this study are EGC-like, but seem to be multipotent rather than pluripotent cells. Nevertheless, the thorough characterization of these cells in this study, and especially the identification of various genes and pathways involved in pluripotency by RNA-Seq, will serve as a rich resource for further derivation of porcine EGC. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. In vitro differentiation of fertile sperm from cryopreserved spermatogonia of the endangered endemic cyprinid honmoroko (Gnathopogon caerulescens)

    Science.gov (United States)

    Higaki, Shogo; Shimada, Manami; Kawamoto, Kazuaki; Todo, Takaaki; Kawasaki, Toshihiro; Tooyama, Ikuo; Fujioka, Yasuhiro; Sakai, Noriyoshi; Takada, Tatsuyuki

    2017-02-01

    Many endemic fish species are threatened with extinction. Conservation strategies and the restoration of endemic fish after extinction must therefore be investigated. Although sperm cryopreservation is indispensable for the conservation of endangered fishes, the limited number of mature fish and limited availability (volume and period) of sperm from small endemic fish hinders the optimization and practical use of this material. In this report, we demonstrate the in vitro differentiation of fertile sperm from cryopreserved spermatogonia of juveniles of the endangered small cyprinid honmoroko (Gnathopogon caerulescens), which is endemic to Lake Biwa in Japan. The entire process of spermatogenesis was recapitulated in vitro using cryopreserved spermatogonia of non-spawning adult and juvenile fish. The differentiation of sperm from spermatogonia was captured as a time-lapse video and confirmed by 5-ethynyl-2‧-deoxyuridine (EdU) incorporation into sperm. Fertility was demonstrated by artificial insemination. These results suggest that the combination of cryopreservation of spermatogonia and in vitro sperm differentiation will provide a new and promising strategy for the preservation of paternal genetic materials.

  17. Clinical predictors of failing one dose of methotrexate for ectopic pregnancy after in vitro fertilization.

    Science.gov (United States)

    Brady, Paula C; Missmer, Stacey A; Farland, Leslie V; Ginsburg, Elizabeth S

    2017-03-01

    The aim of this study is to investigate the clinical predictors of failure of a single dose of methotrexate (MTX) for management of ectopic pregnancy after in vitro fertilization (IVF). A retrospective cohort study was performed of women who conceived ectopic pregnancies following fresh or frozen IVF cycles at an academic infertility clinic between 2007 and 2014, and received intramuscular MTX (50 mg/m2). Successful single-dose MTX treatment was defined as a serum beta-human chorionic gonadotropin (hCG) decline ≥15% between days 4 and 7 post-treatment. Logistic regression models adjusted for oocyte age, number of embryos transferred, and prior ectopic pregnancy were used to estimate the adjusted odds ratio (OR) (95% confidence interval [CI]) of failing one dose of MTX. Sixty-four patients with ectopic pregnancies after IVF were included. Forty required only one dose of MTX (62.5%), while 15 required additional MTX alone (up to four total doses, 23.4%), and 9 required surgery (14.1%). By multivariable logistic regression, the highest tertiles of serum hCG at peak (≥499 IU/L, OR = 9.73, CI 1.88-50.25) and at first MTX administration (≥342 IU/L, OR = 4.74, CI 1.11-20.26), fewer embryos transferred (OR = 0.37 per each additional embryo transferred, CI 0.19-0.74), and adnexal mass by ultrasound (OR = 3.65, CI 1.10-12.11) were each correlated with greater odds of requiring additional MTX and/or surgery. This is the first study to report that in women with ectopic pregnancies after IVF, higher hCG-though well below treatment failure thresholds previously described in spontaneous pregnancies-fewer embryos transferred, and adnexal masses are associated with greater odds of failing one dose of MTX. These findings can be used to counsel IVF patients regarding the likelihood of success with single-dose MTX.

  18. Impact of blood hypercoagulability on in vitro fertilization outcomes: a prospective longitudinal observational study.

    Science.gov (United States)

    Gerotziafas, Grigoris T; Van Dreden, Patrick; Mathieu d'Argent, Emmanuelle; Lefkou, Eleftheria; Grusse, Matthieu; Comtet, Marjorie; Sangare, Rabiatou; Ketatni, Hela; Larsen, Annette K; Elalamy, Ismail

    2017-01-01

    Blood coagulation plays a crucial role in the blastocyst implantation process and its alteration may be related to in vitro fertilization (IVF) failure. We conducted a prospective observational longitudinal study in women eligible for IVF to explore the association between alterations of coagulation with the IVF outcome and to identify the biomarkers of hypercoagulability which are related with this outcome. Thirty-eight women eligible for IVF (IVF-group) and 30 healthy, age-matched women (control group) were included. In the IVF-group, blood was collected at baseline, 5-8 days after administration of gonadotropin-releasing hormone agonist (GnRH), before and two weeks after administration of human follicular stimulating hormone (FSH). Pregnancy was monitored by measurement of β HCG performed 15 days after embryo transfer. Thrombin generation (TG), minimal tissue factor-triggered whole blood thromboelastometry (ROTEM®), procoagulant phospholipid clotting time (Procoag-PPL®), thrombomodulin (TMa), tissue factor activity (TFa), factor VIII (FVIII), factor von Willebrand (FvW), D-Dimers and fibrinogen were assessed at each time point. Positive IVF occurred in 15 women (40%). At baseline, the IVF-group showed significantly increased TG, TFa and TMa and significantly shorter Procoag-PPL versus the control group. After initiation of hormone treatment TG was significantly higher in the IVF-positive as compared to the IVF-negative group. At all studied points, the Procoag-PPL was significantly shorter and the levels of TFa were significantly higher in the IVF-negative group compared to the IVF-positive one. The D-Dimers were higher in the IVF negative as compared to IVF positive group. Multivariate analysis retained the Procoag-PPL and TG as predictors for the IVF outcome. Diagnosis of women with hypercoagulability and their stratification to risk of IVF failure using a model based on the Procoag-PPL and TG is a feasible strategy for the optimization of IVF efficiency

  19. Effect of endometrial cavity fluid on pregnancy rate of fresh versus frozen In Vitro fertilization cycle

    Directory of Open Access Journals (Sweden)

    Nitika Gupta

    2017-01-01

    Full Text Available Objective: This study aims to study the difference in etiology and outcome in terms of implantation rate and abortion rate in fresh (self-stimulated versus frozen (oocyte donation cycle in vitro fertilization (IVF and in transient versus persistent fluid. Material and Methods: This retrospective study was conducted in the Department of Reproductive Medicine of tertiary care center from January 2012 to November 2015. Data were collected retrospectively from the departmental files. Twenty-four patients from fresh IVF-stimulated cycles and 24 from frozen oocyte donation cycle with their endometrium prepared by hormone replacement treatment were included in the study. All patients selected in the study had grade-A embryo transfer of day 3–4 with maximum three embryo transferred. Pregnancy was defined by rising serum beta-human chorionic gonadotrophin levels performed after 14 days of embryo transfer and further confirmed by ultrasonographic visualization of gestational sac at 6 weeks. All biochemical pregnancies were included in implantation failure. All pregnant patients were followed till the termination of pregnancy and further noted as live birth or abortion. Results: Clinical pregnancy rate was seen more in self-stimulated cycle (62.5% with live birth rate of 50% than hormone replacement treatment cycle, in which clinical pregnancy rate was 45.83% with live birth rate of 33.33%. Clinical pregnancy rate was highest in group with very less fluid in cavity (1–2 mm 63% and with live birth of 52.63%. Clinical pregnancy was seen only in two patients of group B with anterior and posterior (AP diameter of fluid in cavity of 2–3 mm with live birth of only one, whereas in group C, with AP diameter of 3–5 mm, none of the patient conceived. This difference was statistically significant. Clinical pregnancy rate was 65.62% in transient fluid accumulation with live birth rate of 53.25%, which was significantly higher than persistent fluid accumulation

  20. 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.

  1. [Deep infiltrative endometriosis without digestive involvement, what is the impact of surgery on in vitro fertilization outcomes? A retrospective study].

    Science.gov (United States)

    Mounsambote, L; Cohen, J; Bendifallah, S; d'Argent, E Mathieu; Selleret, L; Chabbert-Buffet, N; Ballester, M; Antoine, J M; Daraï, E

    2017-01-01

    To evaluate the impact of complete removal of endometriosis in case of deep infiltrative endometriosis without digestive involvement, on in vitro fertilization outcomes. Retrospective monocentric study. We included infertile women with deep infiltrative endometriosis without colorectal involvement that underwent IVF. Women were divided in two groups, following their history: "surgery" when they underwent complete endometriosis resection before IVF and "without surgery" when they underwent IVF without endometriosis removal. We analysed IVF outcomes considering pregnancy rates per cycle and cumulative pregnancy rates per patient. We included 72 patients: 35 in the "surgery" group and 37 in the "without surgery" group. Women in the two groups were comparable in terms of baseline characteristics (age, body mass index, anti-Müllerian hormone, antral follicular count), endometriosis localizations and in vitro fertilization parameters. Cumulative pregnancy rates per patient were similar in both groups (40 % in the "surgery" group and 41 % in the "without surgery" group; P=1). Clinical pregnancy rate per cycle were also comparable groups (24 % in the "surgery" group and 28 % in the "without surgery" group; P=0.67). Surgery performed was comparable in women that became pregnant and in women that did not. Age was lower in women that became pregnant (P=0.01) and there were more pregnancy obtained in women under 35 years. In women with deep infiltrative endometriosis without digestive involvement, in vitro fertilization outcomes were not impacted by surgery. Therapeutic choice between IVF or surgery as first-line treatment remains thus questionable and shall be guided by other influencing factors, such as pain symptomatology, age, tubal permeability, ovarian reserve, partner's sperm characteristics and woman's choice. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  2. Stressful life events are associated with a poor in vitro fertilization (IVF) - outcome: a prospective study

    DEFF Research Database (Denmark)

    Ebbesen, Signe Maria Schneevoigt; Zachariae, Robert; Mehlsen, Mimi Yung

    2009-01-01

    BACKGROUND: There is preliminary evidence to suggest an impact of stress on chances of achieving a pregnancy with in-vitro fertilization (IVF). The majority of the available research has focused on stress related to infertility and going through IVF-treatment, and it is still unclear whether non...... or treatment procedure, including duration of infertility, number of oocytes retrieved and infertility etiology. Mediation analyses indicated that the association between negative life events and IVF pregnancy was partly mediated by the number of oocytes harvested during oocyte retrieval. CONCLUSION: A large...

  3. Biopsy of embryos produced by in vitro fertilization affects development in C57BL/6 mouse strain

    OpenAIRE

    Sugawara, Atsushi; Ward, Monika A.

    2012-01-01

    Preimplantation genetic diagnosis (PGD) is considered highly successful in respect to its accuracy in detecting genetic anomalies but the effects of embryo biopsy on embryonic/fetal growth and development are less known, particularly in conjunction with in vitro fertilization (IVF). Here, we compared biopsied (B) and non-biopsied (NB) mouse embryos for their developmental competence. Embryos C57BL/6 (B6) and B6D2F2 (F2) generated by IVF were subjected to single blastomere biopsy at the 4-cell...

  4. [Jugular vein thrombosis caused by hypercoagulability following in-vitro fertilization-activated protein C resistance and immobilization].

    Science.gov (United States)

    Stölzel, K; Jovanovic, S; Albers, A E

    2013-03-01

    Jugular vein thrombosis (JVT) is extremely difficult to diagnose clinically because of its rarity, the wide range of possible symptoms and the variety of differential diagnoses. A rapid diagnosis is important in order to avoid or prevent imminent life-threatening complications. This study reports a clinical case of extensive JVT due to increased thrombophilia in conjunction with ovarian hyperstimulation syndrome (OHSS) after in vitro fertilization, increased APC resistance and immobilization. It also discusses the current literature that forms the basis for recommendations regarding the diagnosis, therapy and interdisciplinary management.

  5. Transcriptome profile of lung dendritic cells after in vitro porcine reproductive and respiratory syndrome virus (PRRSV) infection.

    Science.gov (United States)

    Pröll, Maren Julia; Neuhoff, Christiane; Schellander, Karl; Uddin, Muhammad Jasim; Cinar, Mehmet Ulas; Sahadevan, Sudeep; Qu, Xueqi; Islam, Md Aminul; Poirier, Mikhael; Müller, Marcel A; Drosten, Christian; Tesfaye, Dawit; Tholen, Ernst; Große-Brinkhaus, Christine

    2017-01-01

    The porcine reproductive and respiratory syndrome (PRRS) is an infectious disease that leads to high financial and production losses in the global swine industry. The pathogenesis of this disease is dependent on a multitude of factors, and its control remains problematic. The immune system generally defends against infectious diseases, especially dendritic cells (DCs), which play a crucial role in the activation of the immune response after viral infections. However, the understanding of the immune response and the genetic impact on the immune response to PRRS virus (PRRSV) remains incomplete. In light of this, we investigated the regulation of the host immune response to PRRSV in porcine lung DCs using RNA-sequencing (RNA-Seq). Lung DCs from two different pig breeds (Pietrain and Duroc) were collected before (0 hours) and during various periods of infection (3, 6, 9, 12, and 24 hours post infection (hpi)). RNA-Seq analysis revealed a total of 20,396 predicted porcine genes, which included breed-specific differentially expressed immune genes. Pietrain and Duroc infected lung DCs showed opposite gene expression courses during the first time points post infection. Duroc lung DCs reacted more strongly and distinctly than Pietrain lung DCs during these periods (3, 6, 9, 12 hpi). Additionally, cluster analysis revealed time-dependent co-expressed groups of genes that were involved in immune-relevant pathways. Key clusters and pathways were identified, which help to explain the biological and functional background of lung DCs post PRRSV infection and suggest IL-1β1 as an important candidate gene. RNA-Seq was also used to characterize the viral replication of PRRSV for each breed. PRRSV was able to infect and to replicate differently in lung DCs between the two mentioned breeds. These results could be useful in investigations on immunity traits in pig breeding and enhancing the health of pigs.

  6. Effect of porcine reproductive and respiratory syndrome virus (PRRSV) (isolate ATCC VR-2385) infection on bactericidal activity of porcine pulmonary intravascular macrophages (PIMs): in vitro comparisons with pulmonary alveolar macrophages (PAMs).

    Science.gov (United States)

    Thanawongnuwech, R; Thacker, E L; Halbur, P G

    1997-11-01

    Porcine pulmonary intravascular macrophages (PIMs) were recovered by in situ pulmonary vascular perfusion with 0.025% collagenase in saline from six 8-week old, crossbred pigs. Pulmonary alveolar macrophages (PAMs) were recovered by bronchoalveolar lavage from the same pigs for comparisons in each assay. The macrophages were exposed to PRRSV (ATCC VR-2385) in vitro for 24 h and infection was confirmed by an indirect immunofluorescence test or transmission electron microscopy. Viral particles tended to accumulate in the vesicles of the Golgi apparatus or endoplasmic reticulum. Bactericidal function assays were performed on the recovered macrophages to determine the effects of the virus on macrophage functions. In vitro PRRSV infection reduced the bactericidal ability of PIMs from 68.3% to 56.4% (P 0.1) at 24 h post-infection. The mean percentage of bacteria killed by macrophages after PRRSV infection was not significantly different among the treatment groups or between the treatment groups and non-infected controls based on colorimetric MTT bactericidal (Staphylococcus aureus) assay. PRRSV did not affect the ability of PIMs or PAMs to internalize opsonized 125I-iododeoxyuridine-labeled S. aureus (P > 0.05). PRRSV infection significantly decreased the production of superoxide anion (P PIMs and by 69.4% in PAMs. PRRSV reduced the myeloperoxidase-H2O2-halide product (P PIMs and by 48.1% for PAMs. The results suggest: (1) PIMs should be considered as an important replication site of PRRSV; (2) PRRSV may have a detrimental effect on both PIMs and PAMs; (3) loss of bactericidal function in PIMs may facilitate hematogenous bacterial infections.

  7. Birth of piglets preselected for gender following in vitro fertilization of in vitro matured pig oocytes by X and Y chromosome bearing spermatozoa sorted by high speed flow cytometry.

    Science.gov (United States)

    Abeydeera, L R; Johnson, L A; Welch, G R; Wang, W H; Boquest, A C; Cantley, T C; Rieke, A; Day, B N

    1998-11-01

    The present study examined the ability to establish pregnancies after transfer of pig embryos derived from in vitro fertilization (IVF) of in vitro matured (IVM) oocytes by X and Y chromosome-bearing spermatozoa sorted by flow cytometry. Cumulus-oocyte complexes (COC) were cultured in BSA-free NCSU-23 medium containing porcine follicular fluid (10%), cysteine (0.1 mg/mL), epidermal growth factor (10 ng/mL), LH (0.5 microgram/mL) and FSH (0.5 microgram/mL) for 22 h, then the oocytes were cultured without hormonal supplements for an additional 22 h. Boar semen was collected and prepared by flow cytometry sorting of X and Y chromosome bearing spermatozoa. After IVM, cumulus-free oocytes were co-incubated with sorted X or Y spermatozoa (2 x 10(4)/mL) for 6 to 7 h in modified Tris-buffered medium containing 2.5 mM caffeine and 0.4% BSA. After IVF, putative embryos were transferred to NCSU-23 medium containing 0.4% BSA for culture. A portion of the oocytes was fixed 12 h after IVF, the remainder were cultured up to 96 h. At 96 h after IVF, 8-cell to morula stage embryos (n = 30 to 35) from each gender were surgically transferred to the uterus of recipient gilts. Insemination of IVM pig oocytes with X- or Y-bearing sperm cells did not influence the rate of penetration (67 vs 80%), polyspermy (40 vs 53%), male pronuclear formation (95 vs 96%), or mean number of spermatozoa per oocyte (1.6 vs 1.6), respectively. Furthermore, no difference was observed between cleavage rates at 48 h after IVF (X, 49 vs Y, 45%). Transfer of embryos derived from X-bearing spermatozoa to 18 recipients resulted in 5 pregnancies and delivery of 23 females and 1 male piglet. Similarly, transfer of embryos derived from Y-bearing sperm cells to 10 recipients resulted in 3 pregnancies, with 9 male piglets delivered. The results show that X- and Y-bearing spermatozoa sorted using USDA sperm sexing technology can be successfully used in an IVM-IVF system to obtain piglets of a predetermined sex.

  8. Management of the first in vitro fertilization cycle for unexplained infertility: a cost-effectiveness analysis of split in vitro fertilization-intracytoplasmic sperm injection.

    Science.gov (United States)

    Vitek, Wendy S; Galárraga, Omar; Klatsky, Peter C; Robins, Jared C; Carson, Sandra A; Blazar, Andrew S

    2013-11-01

    To determine the cost-effectiveness of split IVF-intracytoplasmic sperm injection (ICSI) for the treatment of couples with unexplained infertility. Adaptive decision model. Academic infertility clinic. A total of 154 couples undergoing a split IVF-ICSI cycle and a computer-simulated cohort of women infertility undergoing IVF. Modeling insemination method in the first IVF cycle as all IVF, split IVF-ICSI, or all ICSI, and adapting treatment based on fertilization outcomes. Live birth rate, incremental cost-effectiveness ratio (ICER). In a single cycle, all IVF is preferred as the ICER of split IVF-ICSI or all ICSI ($58,766) does not justify the increased live birth rate (3%). If two cycles are needed, split IVF/ICSI is preferred as the increased cumulative live birth rate (3.3%) is gained at an ICER of $29,666. In a single cycle, all IVF was preferred as the increased live birth rate with split IVF-ICSI and all ICSI was not justified by the increased cost per live birth. If two IVF cycles are needed, however, split IVF/ICSI becomes the preferred approach, as a result of the higher cumulative live birth rate compared with all IVF and the lesser cost per live birth compared with all ICSI. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  9. Management of the first in vitro fertilization cycle for unexplained infertility: a cost-effectiveness analysis of split in vitro fertilization-intracytoplasmic sperm injection

    Science.gov (United States)

    Vitek, Wendy S.; Galárraga, Omar; Klatsky, Peter C.; Robins, Jared C.; Carson, Sandra A.; Blazar, Andrew S.

    2015-01-01

    Objective To determine the cost-effectiveness of split IVF-intracytoplasmic sperm injection (ICSI) for the treatment of couples with unexplained infertility. Design Adaptive decision model. Setting Academic infertility clinic. Patient(s) A total of 154 couples undergoing a split IVF-ICSI cycle and a computer-simulated cohort of women <35 years old with unexplained infertility undergoing IVF. Intervention(s) Modeling insemination method in the first IVF cycle as all IVF, split IVF-ICSI, or all ICSI, and adapting treatment based on fertilization outcomes. Main Outcome Measure(s) Live birth rate, incremental cost-effectiveness ratio (ICER). Result(s) In a single cycle, all IVF is preferred as the ICER of split IVF-ICSI or all ICSI ($58,766) does not justify the increased live birth rate (3%). If two cycles are needed, split IVF/ICSI is preferred as the increased cumulative live birth rate (3.3%) is gained at an ICER of $29,666. Conclusion(s) In a single cycle, all IVF was preferred as the increased live birth rate with split IVF-ICSI and all ICSI was not justified by the increased cost per live birth. If two IVF cycles are needed, however, split IVF/ICSI becomes the preferred approach, as a result of the higher cumulative live birth rate compared with all IVF and the lesser cost per live birth compared with all ICSI. PMID:23876534

  10. Antioxidant effect of crocin on bovine sperm quality and in vitro fertilization.

    Science.gov (United States)

    Sapanidou, V; Taitzoglou, I; Tsakmakidis, Ι; Kourtzelis, I; Fletouris, D; Theodoridis, A; Zervos, I; Tsantarliotou, M

    2015-11-01

    Reactive oxygen species (ROS) production above critical levels affects the genetic and functional integrity of spermatozoa by causing oxidative stress. Spermatozoa are susceptible to oxidative stress in terms of motility and fertilization capacity. Crocin (crocetin di-gentiobiose ester), a main constituent of Crocus Sativus L. (saffron), is known for its antioxidant activity by scavenging ROS, especially superoxide anion. The aim of the present study is to evaluate the effect of crocin on the quality characteristics of spermatozoa and fertilization rate. Frozen-thawed and washed spermatozoa from four different bulls were incubated with three different concentrations of crocin (0.5, 1, and 2 mM), for 120 and 240 minutes, in the presence of a negative control, and were evaluated in terms of motility, viability, acrosomal status, DNA fragmentation index, intracellular ROS, and lipid peroxidation. The most potent concentration of crocin (1 mM) was also added in the fertilization medium to test its impact on fertilization outcome. The results indicate that the incubation of spermatozoa with 1 mM of crocin resulted in a statistically significant lower production of ROS, lower lipid peroxidation and in better maintenance of motility, viability, and acrosomal integrity, with a very small number of fragmented cells, compared to the control and the other treated groups (P quality and its fertilization capability, directly and/or indirectly, by modulating ROS concentration. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Effect of sperm cryopreservation and treatment with calcium ionophore or heparin on in vitro fertilization of horse oocytes.

    Science.gov (United States)

    Alm, H; Torner, H; Blottner, S; Nürnberg, G; Kanitz, W

    2001-09-15

    Little information is available on methods of sperm capacitation for IVF in the horse. In this study, we summarized results of several independent trials that compared acrosome reaction, hyperactivation and chromatin integrity of fresh or cryopreserved stallion spermatozoa after treatment with heparin or with calcium ionophore. We also examined the influence of spermatozoa storage (fresh vs. cryopreserved), capacitation treatment, oocyte maturation time and cumulus morphology on the penetration rate and fertilization rate. We recovered cumulus-oocyte-complexes (COCs) from ovaries by ultrasound guided follicle aspiration or by scraping of follicles from ovaries obtained at a slaughterhouse. Upon recovery, we evaluated the cumulus morphology, and the COCs were matured in vitro for 18 to 24 or 26 to 40 h. Fresh semen and cryopreserved semen were treated either with heparin (200 microg/mL) or calcium ionophore (7.14 microM). Overall, 28.4% (99/349) of the oocytes were penetrated, and 12.9% (45/349) were fertilized. Fresh spermatozoa treated with calcium ionophore showed a higher penetration rate than cryopreserved spermatozoa (36.0 vs. 0%). Fresh and heparin-treated spermatozoa showed a penetration rate of 29.1%, and the same treatment for cryopreserved spermatozoa showed a penetration rate of 33.7%; none of these differences was significant (P>0.05). Fertilization rates after the calcium and heparin treatment followed the same trend and also showed no significant differences. Prolonged maturation period resulted in higher penetration (Pfertilization rates in compact (26 to 40 h: 37.7 and 13.1% vs. 18 to 24 h: 13.1 and 2.8%) and in tendency in expanded COCs (26 to 40 h: 40.0 and 30.3% vs. 18 to 24 h: 29.4 and 13.5%). In oocytes with only a few cumulus cells, the rates tended to be higher after the shorter incubation (18 to 24 h: 33.5 and 18.8% vs. 26 to 40 h: 17.2 and 6.5%). We observed hyperactivation more frequently in fresh than in cryopreserved semen after

  12. Resveratrol prevents capacitation-like changes and improves in vitro fertilizing capability of buffalo frozen-thawed sperm.

    Science.gov (United States)

    Longobardi, Valentina; Zullo, Gianluigi; Salzano, Angela; De Canditiis, Carolina; Cammarano, Andrea; De Luise, Luca; Puzio, Maria Valeria; Neglia, Gianluca; Gasparrini, Bianca

    2017-01-15

    The aim of this study was to evaluate the effect of resveratrol supplementation of semen extender on fertility parameters of frozen-thawed buffalo sperm. After the initial semen assessment, buffalo semen was cryopreserved in BioXcell containing 0 (control group), 0.5, 1, 10, and 50-μM resveratrol. After thawing, viability, motility, and capacitation status (assessed by localization of phosphotyrosine-containing proteins) were evaluated. Based on the results of the dose-response trial, the concentration of 50 μM was selected for further assessments, such as membrane integrity, total antioxidant capacity, reactive oxygen species, and lipid peroxidation (LPO) levels. Moreover, in vitro fertilizing ability by heterologous IVF and in vivo fertility were assessed. No differences among groups were recorded in sperm motility and viability (on average 52.3 ± 2.1% and 76.6 ± 1.3%, respectively). However, data showed a resveratrol dose-dependent effect on sperm capacitation status, with a significant reduction of the cryopreservation-induced capacitation with the higher concentrations tested. In particular, both 10- and 50-μM resveratrol increased (P resveratrol also decreased (P resveratrol increased membrane integrity, indicated by the higher percentage of hypo-osmotic swelling positive sperm (55.6 ± 0.6 vs. 48.4 ± 0.7; P resveratrol-treated sperm compared with the control, as indicated by dihydroethidium values (0.17 ± 0.01 and 0.22 ± 0.01 μM/μL dihydroethidium, respectively; P resveratrol, the normal fertilization rate considerably improved (60.8%, P resveratrol-treated semen (50 μM) compared with the control (48.7 vs. 46.5%, respectively). In conclusion, the inclusion of 50-μM resveratrol in the extender decreases capacitation-like changes and oxidative stress, improving membrane stability and in vitro fertilizing ability of buffalo semen. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Autism and mental retardation among offspring born after in vitro fertilization.

    Science.gov (United States)

    Sandin, Sven; Nygren, Karl-Gösta; Iliadou, Anastasia; Hultman, Christina M; Reichenberg, Abraham

    2013-07-03

    Between 1978 and 2010, approximately 5 million infants were born after in vitro fertilization (IVF) treatments. Yet limited information on neurodevelopment after IVF exists, especially after the first year of life. To examine the association between use of any IVF and different IVF procedures and the risk of autistic disorder and mental retardation in the offspring. A population-based, prospective cohort study using Swedish national health registers. Offspring born between 1982 and 2007 were followed up for a clinical diagnosis of autistic disorder or mental retardation until December 31, 2009. The exposure of interest was IVF, categorized according to whether intracytoplasmic sperm injection (ICSI) for male infertility was used and whether embryos were fresh or frozen. For ICSI, whether sperm were ejaculated or surgically extracted was also considered. Relative risks (RRs) for autistic disorder and mental retardation and rates per 100,000 person-years, comparing spontaneously conceived offspring with those born after an IVF procedure and comparing 5 IVF procedures used in Sweden vs IVF without ICSI with fresh embryo transfer, the most common treatment. We also analyzed the subgroup restricted to singletons. Of the more than 2.5 million infants born, 30,959 (1.2%) were conceived by IVF and were followed up for a mean 10 (SD, 6) years. Overall, 103 of 6959 children (1.5%) with autistic disorder and 180 of 15,830 (1.1%) with mental retardation were conceived by IVF. The RR for autistic disorder after any procedure compared with spontaneous conception was 1.14 (95% CI, 0.94-1.39; 19.0 vs 15.6 per 100,000 person-years). The RR for mental retardation was 1.18 (95% CI, 1.01-1.36; 46.3 vs 39.8 per 100,000 person-years). For both outcomes, there was no statistically significant association when restricting analysis to singletons. Compared with IVF without ICSI with fresh embryo transfer, there were statistically significantly increased risks of autistic disorder following

  14. A novel approach to quantifying ovarian cell lipid content and lipid accumulation in vitro by confocal microscopy in lean women undergoing ovarian stimulation for in vitro fertilization (IVF).

    Science.gov (United States)

    Singh, Prapti; Amin, Marli; Keller, Erica; Simerman, Ariel; Aguilera, Paul; Briton-Jones, Christine; Hill, David L; Abbott, David H; Chazenbalk, Gregorio; Dumesic, Daniel A

    2013-06-01

    To quantify intracellular lipid levels in cumulus cells (CCs) and mural granulosa cells (MGCs) of lean women undergoing gonadotropin therapy for in vitro fertilization (IVF), based upon different cell preparation methods. CCs and MGCs from 16 lean women undergoing ovarian stimulation for IVF were studied. Cells were pooled by cell type, with each type of cell separated into two groups for determination of initial lipid content (Method 1) and subsequent lipid accumulation in vitro (Method 2). Cells for initial lipid content were immediately fixed at the time of the oocyte retrieval with 4% paraformaldehyde in suspension, while those for subsequent lipid accumulation in vitro were cultured for 4 h with 5% fetal calf serum and then fixed. Cells were treated with lipid fluorescent dye BODIPY® FL C16 and nuclear marker DAPI. Intracellular lipid was quantified by confocal microscopy, using ImageJ software analysis. There was no significant effect of cell type (P = 0.2) or cell type-cell preparation method interaction (P = 0.8) on cell area (Method 1: CC 99.7 ± 5.1, MGC 132.8 ± 5.8; Method 2: CC 221.9 ± 30.4, MGC 265.1 ± 48.5 μm(2)). The mean area of all cells combined was significantly less for cells prepared by Method 1 (116.2 ± 4.9 μm(2)) vs. Method 2 (243.5 ± 22.5 μm(2), P vitro was significantly higher in CC (154.0 ± 9.1) than MGC (104.6 ± 9.9 fluorescence/cell area, P vitro (P vitro over time.

  15. Inheritance of a Ring Chromosome 21 in a Couple Undergoing In Vitro Fertilization (IVF: A Case Report

    Directory of Open Access Journals (Sweden)

    Roberto L. P. Mazzaschi

    2011-01-01

    Full Text Available An amniotic fluid sample from an in vitro fertilized pregnancy was referred for cytogenetic analysis based on a Down syndrome screening risk of 1 : 21. Routine cytogenetic analysis showed a nonmosaic karyotype of 46,XX,r(21(p11.2q22.3, with partial monosomy for chromosome 21 due to a ring chromosome replacing one of the normal homologues. Detailed ultrasound scanning for the remainder of the pregnancy did not reveal any unusual findings. Parental bloods showed that the mother was mosaic for the ring 21 with a karyotype of 46,XX,r(21(p11.2q22.3/46,XX and the father had an unrelated Robertsonian translocation, with a karyotype of 45,XY,rob(13;14(q10;q10. Microarray analysis of cultured amniocytes determined the extent of the deletion of chromosome 21 material in the ring. The parents were given genetic counselling, and a phenotypically normal female baby was delivered at term. This case highlights the importance of karyotyping as an initial step in the management of couples referred for in vitro fertilization.

  16. The new system of shorter porcine oocyte in vitro maturation (18 hours) using ≥8 mm follicles derived from cumulus-oocyte complexes.

    Science.gov (United States)

    Kwak, Seong-Sung; Yoon, Junchul David; Cheong, Seung-A; Jeon, Yubyeol; Lee, Eunsong; Hyun, Sang-Hwan

    2014-01-15

    Despite recent efforts to improve in vitro maturation (IVM) systems for porcine oocytes, developmental competence of in vitro-matured oocytes is still suboptimal compared with those matured in vivo. In this study, we compared oocytes obtained from large (≥8 mm; LF) and medium (3-7 mm; MF) sized follicles in terms of nuclear maturation, intracellular glutathione and reactive oxygen species levels, gene expression, and embryo developmental competence after IVM. In the control group, cumulus-oocyte complexes (COCs) were aspirated from MF and matured for 22 hours with hormones and subsequently matured for 18 to 20 hours without hormones at 39 °C, 5% CO2 in vitro. In the LF group, COCs were obtained from follicles larger than 8 mm and were subjected to IVM for only 18 hours. The ovaries have LF were averagely obtained with 1.7% per day during 2012 and it was significantly higher in the winter season. The results of the nuclear stage assessment of the COCs from the LFs are as follows: before IVM (0 hours); germinal vesicle stage (15.2%), metaphase I (MI) stage (55.4%), anaphase and telophase I stages (15.8%), and metaphase II (MII) stage (13.6%). After 6 hours IVM; germinal vesicle (4.2%), MI (43.6%), anaphase and telophase I (9.4%), and MII (42.8%). After 18-hour IVM; MI (9.7%) and MII (90.3%). Oocytes from LF showed a significant (P numbers in blastocysts) compared with the control group. In conclusion, oocytes from LFs require only 18 hours to complete oocyte maturation in vitro and their developmental competence is significantly greater than those obtained from MFs. Although their numbers are limited, oocytes from LFs might offer an alternative source for the efficient production of transgenic pigs using SCNT. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Angiopoietin-2 1087G > A rs3020221 gene polymorphism is associated with in vitro fertilization and embryo transfer outcome

    Directory of Open Access Journals (Sweden)

    Negar Ajabi

    2017-12-01

    Full Text Available In in vitro fertilization (IVF the eggs are fertilized in the laboratory. The success of IVF depends on the angiogenesis. Angiopoetin-2 (ANGPT2 plays a critical role in angiogenesis and acts as a natural antagonist of the endothelial cell–specific receptor tyrosine kinase (Tie-2. A functional single-nucleotide polymorphism (SNP in the ANGPT2 1087 G > A gene is known to influence in the gene expression in an allele-specific manner. The aim of this project was to study the effect of angiopoietin-2 gene polymorphism on in vitro fertilization and embryo transfer (IVF-ET in Iran. The prevalence of G/A substitution in exon 4 was determined in 160 infertile patients enrolled who had unsuccessful IVF history and 160 control subjects who had successful IVF history. The genomic DNA was extracted from the whole blood using DNA extraction techniques. Genotype of ANGPT2 1087 G > A was performed using Allele-specific polymerase chain reaction (AS-PCR. The genotype frequencies of GG, AG and AA in the IVF negative were 7.5%, 67.5% and 25%, respectively and in the IVF positive were 15%, 77.5% and 7.5%, respectively. The allele frequencies of G, A in the IVF negative were 41.2%, 58.8%, respectively and in IVF positive were 53.8%, 46.2%, respectively. Statistical analysis showed that there is a significant correlation in the genotype between two groups (OR = 4.11, 95% CI = 2.065–8.18, P = 0.0001. It is concluded that ANGPT2 G/A polymorphism is associated with IVF-ET outcome in a population in the north of Iran.

  18. Effect of Saccharomyces cerevisiae var. Boulardii and β-galactomannan oligosaccharide on porcine intestinal epithelial and dendritic cells challenged in vitro with Escherichia coli F4 (K88

    Directory of Open Access Journals (Sweden)

    Badia Roger

    2012-01-01

    Full Text Available Abstract Probiotic and prebiotics, often called "immune-enhancing" feed additives, are believed to deal with pathogens, preventing the need of an immune response and reducing tissue damage. In this study, we investigated if a recently developed β-galactomannan (βGM had a similar protective role compared to Saccharomyces cerevisiae var. Boulardii (Scb, a proven probiotic, in the context of enterotoxigenic Escherichia coli (ETEC infection. ETEC causes inflammation, diarrhea and intestinal damage in piglets, resulting in large economic loses worldwide. We observed that Scb and βGM products inhibited in vitro adhesion of ETEC on cell surface of porcine intestinal IPI-2I cells. Our data showed that Scb and βGM decreased the mRNA ETEC-induced gene expression of pro-inflammatory cytokines TNF-α, IL-6, GM-CSF and chemokines CCL2, CCL20 and CXCL8 on intestinal IPI-2I. Furthermore, we investigated the putative immunomodulatory role of Scb and βGM on porcine monocyte-derived dendritic cells (DCs per se and under infection conditions. We observed a slight up-regulation of mRNA for TNF-α and CCR7 receptor after co-incubation of DC with Scb and βGM. However, no differences were found in DC activation upon ETEC infection and Scb or βGM co-culture. Therefore, our results indicate that, similar to probiotic Scb, prebiotic βGM may protect intestinal epithelial cells against intestinal pathogens. Finally, although these products may modulate DC activation, their effect under ETEC challenge conditions remains to be elucidated.

  19. Effect of gibberellic acid on the quality of sperm and in vitro fertilization outcome in adult male rats

    Directory of Open Access Journals (Sweden)

    Mohammadreza Hosseinchi

    2014-12-01

    Full Text Available Gibberellic acid (GA3 is a group of plant hormones identified in various plants. The aim of this study was to determine the effects of GA3 on sperm parameters and in vitro fertilization (IVF. Fifty six adult male rats were divided into seven groups as, control, treatment and sham. Following 15, 30 and 45 days of GA3 and methanol alcohol (MA administration, rats were euthanized and epididymis tail was transferred to human tubular fluid (HTF medium containing 4 mg mL-1 bovine serum albumin (BSA .Total number of sperms, the percentage of live sperms, immature sperms and sperms with damaged chromatin and IVF were examined. The oocytes were obtained from immature rats after the injection of pregnant mare's serum (PMSG and human chorionic gonadotropin (HCG hormones. Human tubular fluid was used as the fertilization medium and zygotes transferred to fresh 1-cell rat embryos culture medium (mR1ECM to reach the blastocyst stage. This study showed that GA3 could decrease the number of total sperms on days 30 and 45 in treated group comparison with the control and sham groups. Additionally, GA3 increased the immature sperms and sperms with damaged chromatin. The percentage of fertilization, two-cell embryos and blastocyst resulting from the treatment group on days 30 and 45 also decreased and showed significant differences with the control and sham groups (p < 0.05. The results obtained from this study indicated that the oral use of GA3 could reduce the fertility in rats by influencing the sperm number and the quality of sperm’s chromatins.

  20. In vitro and in vivo degradation of rapamycin-eluting Mg-Nd-Zn-Zr alloy stents in porcine coronary arteries.

    Science.gov (United States)

    Shi, Yongjuan; Zhang, Lei; Chen, Jiahui; Zhang, Jian; Yuan, Feng; Shen, Li; Chen, Chenxin; Pei, Jia; Li, Zhonghua; Tan, Jinyun; Yuan, Guangyin

    2017-11-01

    In this work, rapamycin-eluting poly (d, l-lactic acid) coating (PDLLA/RAPA) was prepared on biodegradable Mg-Nd-Zn-Zr alloy (JDBM) for both in vitro and in vivo investigation of the degradation behaviors of the magnesium alloy stent platform. Electrochemical tests and hydrogen evolution test demonstrated significant in vitro protection of the polymeric coating against magnesium degradation both in short and long term. The 3-month in vivo study on the RAPA-eluting JDBM stent implanted into porcine coronary arteries confirmed its favorable safety, and in the meanwhile revealed similar neointima proliferation compared to the second generation DES Firebird 2 with no occurrence of adverse complications. Moreover, Micro-CT examination combined with IVUS and OCT detection indicated a remarkably lower degradation rate and prolonged radial supporting duration of the drug-eluting JDBM stent as compared to the bare, attributable to the protection of the coating in vivo. Hence, rapamycin-eluting JDBM stents exhibit great potential for clinical application. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Bottlenose Dolphin (Tursiops truncatus) Spermatozoa: Collection, Cryopreservation, and Heterologous In Vitro Fertilization.

    Science.gov (United States)

    Sánchez-Calabuig, María Jesús; García-Vázquez, Francisco Alberto; Laguna-Barraza, Ricardo; Barros-García, Carlos; García-Parraga, Daniel; Rizos, Dimitrios; Gutiérrez Adan, Alfonso; Pérez-Gutíerrez, José Félix

    2017-08-21

    The use of cryopreserved dolphin spermatozoa facilitates the exchange of genetic material between aquatic parks and makes spermatozoa accessible to laboratories for studies to further our understanding of marine mammal reproduction. Heterologous IVF, a replacement for homologous IVF, could provide a means to test the sperm fertility potential; to study gamete physiology and early embryo development; and to avoid the use of valuable dolphin oocytes, which are difficult to obtain. Here, we present protocols that have been successfully used to collect and cryopreserve dolphin spermatozoa. The collection of semen is performed by manual stimulation on trained dolphins. Cryopreservation is accomplished using a TRIS egg-yolk based extender with glycerol. In addition, we present a protocol that describes heterologous IVF using dolphin spermatozoa and bovine oocytes and that verifies the hybrid nature of the resulting embryo using PCR. Heterologous fertilization raises questions on fertilization and can be used as a tool to study gamete physiology and early embryo development. In addition, the success of heterologous IVF demonstrates the potential of this technique to test dolphin sperm fertilizing capacity, which is worth further examination.

  2. Stages of endometriosis: Does it affect in vitro fertilization outcome

    Directory of Open Access Journals (Sweden)

    Sonja Pop-Trajkovic

    2014-06-01

    Conclusion: The American Society for Reproductive Medicine classification of endometriosis is useful in predicting IVF outcome. Advanced endometriosis means a worse prognosis for IVF treatment compared to milder stages or tubal factor infertility. The decreased fertilization rate in Stage I/II endometriosis might be a cause of subfertility in these women, as a result of a hostile environment caused by the disease.

  3. LOCALIZATION OF THE SPERM PROTEIN SP22 AND INHIBITION OF FERTILITY IN VIVO AND IN VITRO

    Science.gov (United States)

    We previously established that the levels sperm membrane protein SP22 are highly correlated with the fertility of sperm from the cauda epididymidis of rats exposed to both epididymal and testicular toxicants, and that a testis-specific SP22 transcript is expressed in post-meiotic...

  4. Optimization of three-dimensional imaging on in vitro produced porcine blastocysts and chimeras for stem cell testing

    DEFF Research Database (Denmark)

    Secher, Jan Ole Bertelsen; Freude, Kristine; Li, Rong

    2015-01-01

    of the ICM they are likely to contribute to the fetus and if they are located in the area of the TE they are likely to contribute to the fetal membranes. In this article, we optimize on methods for embryo staining and mounting so that the exact location of injected stem cells within preimplantation porcine......Differential staining is an immunocytochemical staining that visualizes trophectoderm (TE) and the inner cell mass (ICM) of the blastocysts. It is used to determine the blastocyst quality, but could also be a useful tool to assess the integration site of injected cells into the early embryo....... This is relevant for testing of presumed pluripotent stem cells. The gold standard for pluripotent stem cells is to test if the cells are capable of contributing to germline chimeras. Differential staining can be used to evaluate the possibility of chimeric contribution; if the cells are located in the area...

  5. In vitro cultivation and partial characterization of Lawsonia intracellularis from a Japanese field case of porcine proliferative enteropathy.

    Science.gov (United States)

    Koyama, Tomohiro; Hirai, Takuya; Nagai, Shinya

    2006-06-01

    Lawsonia intracellularis isolated from a Japanese field case of porcine proliferative enteropathy (PPE) was cultivated and partially characterized. The bacterial cells isolated from the intestinal mucosa of a pig suffering from the acute form of PPE were used to inoculate rat small intestine cells (IEC-18) and human epithelial cells (HEp-2). Infected foci, which were stained with L. intracellularis-specific rabbit antiserum, were observed in the cell culture at 5 days post inoculation. The DNA sequence of several genes in the Japanese isolate had high similarity with those of the L. intracellularis type strain, suggesting the genetically close relationship of the two strains. This is the first report describing the cultivation and partial characterization of L. intracellularis originated in Japan.

  6. Transcriptome profile of lung dendritic cells after in vitro porcine reproductive and respiratory syndrome virus (PRRSV) infection

    DEFF Research Database (Denmark)

    Pröll, Maren Julia; Neuhoff, Christiane; Schellander, Karl

    2017-01-01

    generally defends against infectious diseases, especially dendritic cells (DCs), which play a crucial role in the activation of the immune response after viral infections. However, the understanding of the immune response and the genetic impact on the immune response to PRRS virus (PRRSV) remains incomplete....... In light of this, we investigated the regulation of the host immune response to PRRSV in porcine lung DCs using RNA-sequencing (RNA-Seq). Lung DCs from two different pig breeds (Pietrain and Duroc) were collected before (0 hours) and during various periods of infection (3, 6, 9, 12, and 24 hours post...... could be useful in investigations on immunity traits in pig breeding and enhancing the health of pigs....

  7. Decolonisation of MRSA, S. aureus and E. coli by cold-atmospheric plasma using a porcine skin model in vitro.

    Directory of Open Access Journals (Sweden)

    Tim Maisch

    Full Text Available In the last twenty years new antibacterial agents approved by the U.S. FDA decreased whereas in parallel the resistance situation of multi-resistant bacteria increased. Thus, community and nosocomial acquired infections of resistant bacteria led to a decrease in the efficacy of standard therapy, prolonging treatment time and increasing healthcare costs. Therefore, the aim of this work was to demonstrate the applicability of cold atmospheric plasma for decolonisation of Gram-positive (Methicillin-resistant Staphylococcus aureus (MRSA, Methicillin-sensitive Staphylococcus aureus and Gram-negative bacteria (E. coli using an ex vivo pig skin model. Freshly excised skin samples were taken from six month old female pigs (breed: Pietrain. After application of pure bacteria on the surface of the explants these were treated with cold atmospheric plasma for up to 15 min. Two different plasma devices were evaluated. A decolonisation efficacy of 3 log(10 steps was achieved already after 6 min of plasma treatment. Longer plasma treatment times achieved a killing rate of 5 log(10 steps independently from the applied bacteria strains. Histological evaluations of untreated and treated skin areas upon cold atmospheric plasma treatment within 24 h showed no morphological changes as well as no significant degree of necrosis or apoptosis determined by the TUNEL-assay indicating that the porcine skin is still vital. This study demonstrates for the first time that cold atmospheric plasma is able to very efficiently kill bacteria applied to an intact skin surface using an ex vivo porcine skin model. The results emphasize the potential of cold atmospheric plasma as a new possible treatment option for decolonisation of human skin from bacteria in patients in the future without harming the surrounding tissue.

  8. Development of a rapid in vitro protein refolding assay which discriminates between peptide-bound and peptide-free forms of recombinant porcine major histocompatibility class I complex (SLA-I)

    DEFF Research Database (Denmark)

    Oleksiewicz, M.B.; Kristensen, B.; Ladekjaer-Mikkelsen, A.S.

    2002-01-01

    , by linking them through glycine-rich linkers to peptides representing T-cell epitopes from classical swine fever virus (CSFV) and foot-and-mouth disease virus (FMDV). An in vitro refold assay was developed, using a monoclonal anti-SLA antibody (PT85A) to gauge refolding. The single best-defined, SLA......-I restricted porcine CD8(+) T-cell epitope currently known is a 9-residue peptide from the polyprotein of CSFV (J. Gen. Virol, 76 (1995) 3039). Based on results with the CSFV epitope and two porcine haplotypes (H4 and H7), the in vitro refold assay appeared able to discriminate between peptide-free and peptide...

  9. Modulation of porcine endogenous retroviruses (PERVs) expression in vitro by shRNA in the presence of cyclosporine A and dexamethasone.

    Science.gov (United States)

    Sypniewski, Daniel; Bednarek, Ilona; Matczyńska, Daria; Gałka, Sabina; Loch, Tomasz; Sołtysik, Dagna; Machnik, Grzegorz; Nowak, Ewa

    2012-12-31

    Despite the fact that the risk of Porcine Endogenous Retroviruses (PERV) infection and propagation in human recipients is extremely low, such an event cannot be completely ruled out, especially in immunosuppressed patients. Therefore, the aim of this study was to analyze the expression of PERVs in vitro in the presence of immunosuppression agents: cyclosporine A (CsA), and dexamethasone (DEX). We investigated the possible interactions between immunosuppression drugs, CsA and DEX, and the efficiency of anti-PERV RNAi. Plasmid-based vectors expressing shRNAs against all PERV genes were constructed and analyzed. PERVs expression in cultures transfected with anti-PERV RNAi constructions and treated with CsA or DEX was analyzed by Real-Time RT-PCR, Western blot, and by the measurement of RT activity. Both CsA and DEX inhibited PERVs expression in cell cultures in vitro. RNAi constructions efficiently knocked down PERV expression in Circe, and de novo PERV-infected HeLa and HEK-293 cell cultures. Pretreatment of Circe cultures with CsA or DEX increased PERVs knockdown by RNAi, but no specific interaction between the drugs and transfection efficiency was observed. Our results demonstrate that cyclosporine A and dexamethasone decrease expression of PERVs in vitro. We also proved that these drugs did not synergize or antagonize RNAi-mediated knockdown of PERVs. These observations may be beneficial in immunosuppressed xenograft recipients; however, due to the controversial literature data concerning influence of immune suppression on graft recipients, our results should be further analyzed.

  10. Characterization of porcine multipotent stem/stromal cells derived from skin, adipose, and ovarian tissues and their differentiation in vitro into putative oocyte-like cells.

    Science.gov (United States)

    Song, Seung-Hee; Kumar, Basavarajappa Mohana; Kang, Eun-Ju; Lee, Yeon-Mi; Kim, Tae-Ho; Ock, Sun-A; Lee, Sung-Lim; Jeon, Byeong-Gyun; Rho, Gyu-Jin

    2011-08-01

    The present study evaluated the alkaline phosphatase activity, cell cycle stage, expression of markers and early transcriptional factors, and in vitro differentiation into selected cell lineages of porcine stem/stromal cells (SCs) isolated from skin (SSCs), adipose, and ovarian (OSCs) tissues. Skin and adipose SCs were isolated from a 6-month-old miniature pig, whereas OSCs were isolated from a newly born piglet. Isolated cells exhibited fibroblast-like cell population with significant renewal capacity and formed colonies by cells out-growth. All cells were positive for alkaline phosphatase activity and showed a relatively lower population at G0/G1 phase of the cell cycle. SCs derived from all tissues were strongly positive for cell surface markers, such as CD29, CD44, CD90, and vimentin. Further, relatively lower expression of cytokeratin and immunophenotype markers, such as major histocompatibility complex II (MHCII) and swine leukocyte antigen (SLA), was also observed. SCs derived from all tissues positively expressed the transcription factors, such as Oct-3/4, Nanog, and Sox-2. After induction, all SCs successfully differentiated into osteocytes and adipocytes and expressed the lineage specific marker genes. Further, cells from all tissues exhibited their potential for in vitro oogenesis with morphological changes and expression of markers during the germ-cell formation, namely Oct-4, growth differentiation factor 9b, c-Mos, Vasa, deleted in azoospermia-like gene, zona pellucida C, and follicle stimulating hormone receptor. Apart from basic features and selected lineage potential among all types of cells, OSCs possessed a greater ability to differentiate into the germ cell lineage in vitro.

  11. Multivariate analyses for determining the association of field porcine fertility with sperm motion traits analysed by computer-assisted semen analysis and with sperm morphology.

    Science.gov (United States)

    Kummer, A B H P; Gaggini, T S; Bernardi, M L; McManus, C; Gonçales, E M; Wentz, I; Bortolozzo, F P

    2013-10-01

    This study investigates the association of semen traits with boar fertility. The fertility outcome (farrowing rate - FR and total piglets born - TB) of 14 boars was obtained from a field trial conducted during 10 week of breeding period on a commercial farm using multiparous sows (n = 948) through single-sire mating with 2 × 10(9) motile sperm cells per artificial insemination (AI) dose. Sperm motion parameters, evaluated with computer-assisted semen analysis system in raw and stored semen at 17°C for 240 h, in addition to morphological sperm defects, measured on the collection day, were included in the analysis to determine which semen traits were important to discriminate the fertility potential of ejaculates from these boars. The data underwent multivariate cluster, canonical and discriminant analyses. Four clusters of boars were formed based on fertility outcome. One boar, with the lowest FR and TB values (89.7% and 11.98), and two boars, with the highest FR and TB values (97.8% and 14.16), were placed in different clusters. The other boars were separated in two distinct clusters (four and seven boars), including boars with intermediate TB (12.64 and 13.22) but divergent values for FR (95.9% vs 91.8%). Semen traits with higher discriminatory power included total motility, progressive motility, amplitude of lateral head displacement and cytoplasmatic droplets. Through multivariate discriminant analysis, more than 80% of the 140 ejaculates were correctly classified into their own group, showing that this analysis may be an efficient statistical tool to improve the discrimination of potential fertility of boars. Nevertheless, the validation of the relationship between fertility and semen traits using this statistical approach needs to be performed on a larger number of farms and with a greater number of boars. © 2013 Blackwell Verlag GmbH.

  12. Use of polarized light microscopy in porcine reproductive technologies.

    Science.gov (United States)

    Caamaño, J N; Maside, C; Gil, M A; Muñoz, M; Cuello, C; Díez, C; Sánchez-Osorio, J R; Martín, D; Gomis, J; Vazquez, J M; Roca, J; Carrocera, S; Martinez, E A; Gómez, E

    2011-09-01

    The meiotic spindle in the oocyte is composed of microtubules and plays an important role during chromosome alignment and separation at meiosis. Polarized light microscopy (PLM) could be useful for a non-invasive evaluation of the meiotic spindle and may allow removal of nuclear structures without fluorochrome staining and ultraviolet exposure. In this study, PLM was used to assess its potential application in porcine reproductive technologies. The objectives of the present study were to assess the efficiency of PLM to detect microtubule-polymerized protein in in vitro-matured porcine oocytes; to examine its effects on the oocyte developmental competence; to select oocytes based on the presence of the meiotic spindle detected by PLM; and to assess the efficiency oocyte enucleation assisted with PLM. In the first experiment, the presence of microtubule-polymerized protein was assessed and confirmed in oocytes (n = 117) by immunostaining and chromatin detection. In the second experiment, oocytes (n = 160) were exposed or not (controls) to PLM for 10 minutes, and then parthenogenetically activated and cultured in vitro. In the third experiment, development competence of oocytes with a positive or negative signal to PLM was analyzed after in vitro fertilization. Finally, oocytes (n = 54) were enucleated using PLM as a tool to remove the meiotic spindle. A positive PLM signal was detected in 98.2 % of the oocytes, which strongly correlated (r = 1; p PLM did not differ significantly from controls on cleavage, total blastocyst, expanded blastocyst rates and total cell numbers. The percentage of oocytes at the MII stage and blastocyst formation rate in the negative PLM group significantly differed from control and PLM positive groups. Overall efficiency of spindle removal using the PLM-Oosight system was 92.6%. These results suggest that polarized light microscopy is an efficient system to detect microtubule-polymerized protein in in vitro-matured porcine oocytes and does

  13. In vitro study of biological activities of anthocyanin-rich berry extracts on porcine intestinal epithelial cells.

    Science.gov (United States)

    Kšonžeková, Petra; Mariychuk, Ruslan; Eliašová, Adriana; Mudroňová, Dagmar; Csank, Tomáš; Király, Ján; Marcinčáková, Dana; Pistl, Juraj; Tkáčiková, L'udmila

    2016-03-15

    Anthocyanins, compounds that represent the major group of flavonoids in berries, are one of the most powerful natural antioxidants. The aim of this study was to evaluate biological activities and comparison of anthocyanin-rich extracts prepared from chokeberry (Aronia melanocarpa), elderberry (Sambucus nigra), bilberry (Vaccinium myrtillus) and blueberry (V. corymbosum) on the porcine intestinal epithelial IPEC-1 cell line. The IC50 values calculated in the antioxidant cell-based dichlorofluorescein assay (DCF assay) were 1.129 mg L(-1) for chokeberry, 1.081 mg L(-1) for elderberry, 2.561 mg L(-1) for bilberry and 2.965 mg L(-1) for blueberry, respectively. We found a significant negative correlation (P < 0.001) between cyanidin glycosides content and IC50 values. Moreover, extracts rich in cyanidin glycosides stimulated proliferation of IPEC-1 cells and did not have cytotoxic effect on cells at an equivalent in vivo concentration. We found that the chokeberry and elderberry extracts rich in cyanidin glycosides possess better antioxidant and anticytotoxic activities in comparison to blueberry or bilberry extracts with complex anthocyanin profiles. © 2015 Society of Chemical Industry.

  14. Tissue engineering of heart valves: PEGylation of decellularized porcine aortic valve as a scaffold for in vitro recellularization.

    Science.gov (United States)

    Zhou, Jianliang; Hu, Shidong; Ding, Jingli; Xu, Jianjun; Shi, Jiawei; Dong, Nianguo

    2013-09-05

    Poly (ethylene glycol) (PEG) has attracted broad interest for tissue engineering applications. The aim of this study was to synthesize 4-arm -PEG-20kDa with the terminal group of diacrylate (4-arm-PEG-DA) and evaluate its dual functionality for decellularized porcine aortic valve (DAV) based on its mechanical and biological properties. 4-arm-PEG-DA was synthesized by graft copolymerization of linear PEG 20,000 monomers, and characterized by IR1H NMR and 13C NMR; PEGylation of DAV was achieved by the Michael addition reaction between propylene acyl and thiol, its effect was tested by uniaxial planar tensile testing, hematoxylin and eosin (HE) and scanning electron microscopy (SEM). Gly-Arg-Gly-Asp-Ser-Pro-Cys (GRGDSPC) peptides and vascular endothelial growth factor-165 (VEGF165) were conjugated onto DAV by branched PEG-DA (GRGDSPC-PEG-DAV-PEG-VEGF165). Mechanical testing confirmed that PEG-cross-linking significantly enhanced the tensile strength of DAV. Immunofluoresce confirmed the GRGDSPC peptides and VEGF165 were conjugated effectively onto DAV; the quantification of conjunction was completed roughly using spectrophotometry and ELISA. The human umbilical vein endothelial cells (HUVECs) grew and spread well on the GRGDSPC-PEG-DAV-PEG-VEGF165. Therefore, PEGylation of DAV not only can improve the tensile strength of DAV, and can also mediate the conjugation of bioactive molecule (VEGF165 and GRGDSPC peptides) on DAV, which might be suitable for further development of tissue engineered heart valve.

  15. In vitro porcine blood-brain barrier model for permeability studies: pCEL-X software pKa(FLUX) method for aqueous boundary layer correction and detailed data analysis.

    Science.gov (United States)

    Yusof, Siti R; Avdeef, Alex; Abbott, N Joan

    2014-12-18

    In vitro blood-brain barrier (BBB) models from primary brain endothelial cells can closely resemble the in vivo BBB, offering valuable models to assay BBB functions and to screen potential central nervous system drugs. We have recently developed an in vitro BBB model using primary porcine brain endothelial cells. The model shows expression of tight junction proteins and high transendothelial electrical resistance, evidence for a restrictive paracellular pathway. Validation studies using small drug-like compounds demonstrated functional uptake and efflux transporters, showing the suitability of the model to assay drug permeability. However, one limitation of in vitro model permeability measurement is the presence of the aqueous boundary layer (ABL) resulting from inefficient stirring during the permeability assay. The ABL can be a rate-limiting step in permeation, particularly for lipophilic compounds, causing underestimation of the permeability. If the ABL effect is ignored, the permeability measured in vitro will not reflect the permeability in vivo. To address the issue, we explored the combination of in vitro permeability measurement using our porcine model with the pKa(FLUX) method in pCEL-X software to correct for the ABL effect and allow a detailed analysis of in vitro (transendothelial) permeability data, Papp. Published Papp using porcine models generated by our group and other groups are also analyzed. From the Papp, intrinsic transcellular permeability (P0) is derived by simultaneous refinement using a weighted nonlinear regression, taking into account permeability through the ABL, paracellular permeability and filter restrictions on permeation. The in vitro P0 derived for 22 compounds (35 measurements) showed good correlation with P0 derived from in situ brain perfusion data (r(2)=0.61). The analysis also gave evidence for carrier-mediated uptake of naloxone, propranolol and vinblastine. The combination of the in vitro porcine model and the software

  16. The predictive value of three-dimensional Doppler ultrasonography in determining implantation in patients underwent in vitro fertilization

    Directory of Open Access Journals (Sweden)

    Yusuf Çakmak

    2015-12-01

    Full Text Available Objective: The aim of this study is to evaluate the relationship between endometrial and sub-endometrial blood flow and implantation rate in patients whose undergone in vitro fertilization. Methods: A prospective study was conducted. Long protocol and antagonist regimens were administered to the patients. Endometrial and sub-endometrial blood flow was evaluated by using 3 dimensional Doppler ultrasonography on the day of oocyt retrieval measurement. For comparison pregnant and non-pregnant women in terms of endometrial and sub-endometrial blood flow, t test was used. The p value was considered statistically meaningful as 0.05. In long protocol group, the number of embryo was greater in pregnant women than non-pregnant women (p=0.012. The number of transferred embryo increased pregnancy rate almost 3.5 fold (p=0.002. Conclusion: The endometrial and subendometrial blood flow is not reliable factor in prediction pregnancy or implantation.

  17. Hair Trace Element and Electrolyte Content in Women with Natural and In Vitro Fertilization-Induced Pregnancy.

    Science.gov (United States)

    Skalny, Anatoly V; Tinkov, Alexey A; Voronina, Irina; Terekhina, Olga; Skalnaya, Margarita G; Kovas, Yulia

    2017-04-25

    The objective of the present study was to perform comparative analysis of hair trace element content in women with natural and in vitro fertilization (IVF)-induced pregnancy. Hair trace element content in 33 women with IVF-induced pregnancy and 99 age- and body mass index-matched control pregnant women (natural pregnancy) was assessed using inductively coupled plasma mass spectrometry. The results demonstrated that IVF-pregnant women are characterized by significantly lower hair levels of Cu, Fe, Si, Zn, Ca, Mg, and Ba at p iron, zinc, calcium, and magnesium deficiency and arsenic overload in women with IVF-induced pregnancy. The obtained data indicate the necessity of regular monitoring of micronutrient status in IVF-pregnant women in order to prevent potential deleterious effects of altered mineral homeostasis.

  18. Live birth rates following in vitro fertilization in women with thyroid autoimmunity and/or subclinical hypothyroidism.

    Science.gov (United States)

    Chai, Joyce; Yeung, Wing-Yee T; Lee, Chi-Yan V; Li, Hang-Wun R; Ho, Pak-Chung; Ng, Hung-Yu E

    2014-01-01

    To investigate whether the live birth rate following in vitro fertilization (IVF) is affected by thyroid autoimmunity (TAI) and/or subclinical hypothyroidism in subfertile women. Retrospective study in a university infertility clinic. A total of 627 women without past or current history of thyroid disorder undergoing their first IVF cycle. Pre-IVF archived blood serum samples were tested for TAI and thyroid function tests. Live birth rate. The clinical pregnancy rate, live birth rate and miscarriage rate were similar among women with or without TAI and/or subclinical hypothyroidism using a TSH threshold 4·5 mIU/l. Thyroid autoantibody level did not affect these IVF outcomes. The live birth rate and miscarriage rate of women with TAI and/or subclinical hypothyroidism following IVF were not impaired. © 2013 John Wiley & Sons Ltd.

  19. GnRH agonist versus GnRH antagonist in in vitro fertilization and embryo transfer (IVF/ET

    Directory of Open Access Journals (Sweden)

    Depalo Raffaella

    2012-04-01

    Full Text Available Abstract Several protocols are actually available for in Vitro Fertilization and Embryo Transfer. The review summarizes the main differences and the clinic characteristics of the protocols in use with GnRH agonists and GnRH antagonists by emphasizing the major outcomes and hormonal changes associated with each protocol. The majority of randomized clinical trials clearly shows that in “in Vitro” Fertilization and Embryo Transfer, the combination of exogenous Gonadotropin plus a Gonadotropin Releasing Hormone (GnRH agonist, which is able to suppress pituitary FSH and LH secretion, is associated with increased pregnancy rate as compared with the use of gonadotropins without a GnRH agonist. Protocols with GnRH antagonists are effective in preventing a premature rise of LH and induce a shorter and more cost-effective ovarian stimulation compared to the long agonist protocol. However, a different synchronization of follicular recruitment and growth occurs with GnRH agonists than with GnRH antagonists. Future developments have to be focused on timing of the administration of GnRH antagonists, by giving a great attention to new strategies of stimulation in patients in which radio-chemotherapy cycles are needed.

  20. The effect of using complementary medicine on the infertility-specific quality of life of women undergoing in vitro fertilization.

    Science.gov (United States)

    Porat-Katz, Anat; Paltiel, Ora; Kahane, Arik; Eldar-Geva, Talia

    2016-11-01

    To evaluate associations between the use of complementary medicine, quality of life (QoL), and lifestyle habits among women undergoing in vitro fertilization (IVF). In a cross-sectional study, women aged 18-44 years undergoing an IVF cycle at a large IVF center in Israel between February 1, 2013 and April 30, 2015 were invited to complete a self-administered questionnaire. Patients who reported using of at least one complementary medicine intervention to treat infertility prior to IVF treatment were considered complementary-medicine users. Fertility QoL and lifestyle behaviors were compared between complementary-medicine users and non-users with the FertiQoL tools. Of 381 patients eligible to participate in the study, 323 completed the questionnaire; 110 (34.1%) participants were complementary-medicine users. Complementary-medicine users demonstrated higher scores for the FertiQol relational domain (P=0.005) and lower scores for the social domain (P=0.010). Complementary-medicine users reported greater utilization of psychosocial support (Pinfertility could be useful in identifying patients who could benefit from psychosocial interventions or lifestyle recommendations. Copyright © 2016 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

  1. Transrectal electroejaculation combined with in-vitro fertilization: effective treatment of anejaculatory infertility due to spinal cord injury.

    Science.gov (United States)

    Brinsden, P R; Avery, S M; Marcus, S; Macnamee, M C

    1997-12-01

    Infertility due to spinal cord injury (SCI) in young men is a frequent complication of their injury. When the simpler methods of management of the erectile and ejaculatory dysfunction that invariably follow the more severe types of SCI are not effective, then semen production by transrectal electroejaculation (TREE) combined with in-vitro fertilization (IVF) and embryo transfer is effective. A retrospective analysis is presented of data on the treatment and outcome of 35 couples who wished to have a family but in whom the male partner had suffered SCI. These 35 couples had 71 attempts at IVF with spermatozoa obtained following TREE. Normal fertilization and cleavage of the embryos occurred in 48.2% of the oocytes. Fresh embryos were transferred in 54 cycles and frozen-thawed embryos in 14 cycles. In all, 18 clinical pregnancies were achieved in 54 fresh and 14 frozen embryo transfer cycles, with a live birth rate of 16.5% (14/85) per treatment cycle started, 20.6% (14/68) per transfer cycle and 40.0% (14/35) per couple who started treatment, in a mean of 1.9 transfer cycles. We conclude that TREE combined with IVF and embryo transfer is an effective treatment for the infertility problems associated with SCI.

  2. Oocyte retrieval timing based on spontaneous luteinizing hormone surge during natural cycle in vitro fertilization treatment.

    Science.gov (United States)

    Bodri, Daniel; Kawachiya, Satoshi; Kondo, Masae; Kato, Ryutaro; Matsumoto, Tsunekazu

    2014-04-01

    To determine the efficiency of oocyte retrieval (OR) timing based on the occurrence of spontaneous LH surge during natural cycle IVF (ncIVF) treatment. Retrospective cohort study. The cohort was divided into five subgroups according to the presumed stage of spontaneous LH surge on scheduling day (1A: before onset; 1B: surge start; 2: ascending slope; 3: peak; and 4: descending slope). Private infertility clinic. Three hundred sixty-five infertile patients who underwent 1,138 ncIVF treatment cycles during 2008-2011. Drug-free ncIVF treatment. Rate of successfully retrieved, fertilized oocytes, cleaved embryos, and live births per scheduled oocyte retrieval. In 61% of the cycles OR was scheduled before or just at the start of the LH surge (groups 1A-1B), whereas in the remaining cases it was scheduled after the surge had already started (groups 2-4). The proportion of cycles with successfully recovered (range, 71%-86%), inseminated (range, 61%-78%), fertilized oocytes (range, 47%-68%), cleaved embryos (range, 45%-66%), and live births (range, 4.1%-9.2%) was not significantly different among subgroups. In ncIVF treatment OR timing based on the occurrence of spontaneous LH surge is feasible, yielding acceptable oocyte recovery, fertilization, and embryo cleavage rates. This strategy combined with a rapid and low-risk OR procedure permits the management of a large ncIVF program on a 7-days-per-week basis within working hours. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  3. Molecular and Cellular Mechanisms of Sperm-Oocyte Interactions Opinions Relative to in Vitro Fertilization (IVF

    Directory of Open Access Journals (Sweden)

    George Anifandis

    2014-07-01

    Full Text Available One of the biggest prerequisites for pregnancy is the fertilization step, where a human haploid spermatozoon interacts and penetrates one haploid oocyte in order to produce the diploid zygote. Although fertilization is defined by the presence of two pronuclei and the extraction of the second polar body the process itself requires preparation of both gametes for fertilization to take place at a specific time. These preparations include a number of consecutive biochemical and molecular events with the help of specific molecules and with the consequential interaction between the two gametes. These events take place at three different levels and in a precise order, where the moving spermatozoon penetrates (a the outer vestments of the oocyte, known as the cumulus cell layer; (b the zona pellucida (ZP; where exocytosis of the acrosome contents take place and (c direct interaction of the spermatozoon with the plasma membrane of the oocyte, which involves a firm adhesion of the head of the spermatozoon with the oocyte plasma membrane that culminates with the fusion of both sperm and oocyte membranes (Part I. After the above interactions, a cascade of molecular signal transductions is initiated which results in oocyte activation. Soon after the entry of the first spermatozoon into the oocyte and oocyte activation, the oocyte’s coat (the ZP and the oocyte’s plasma membrane seem to change quickly in order to initiate a fast block to a second spermatozoon (Part II. Sometimes, two spermatozoa fuse with one oocyte, an incidence of 1%–2%, resulting in polyploid fetuses that account for up to 10%–20% of spontaneously aborted human conceptuses. The present review aims to focus on the first part of the human sperm and oocyte interactions, emphasizing the latest molecular and cellular mechanisms controlling this process.

  4. Sericin accelerates the production of hyaluronan and decreases the incidence of polyspermy fertilization in bovine oocytes during in vitro maturation.

    Science.gov (United States)

    Hosoe, Misa; Yoshida, Nao; Hashiyada, Yutaka; Teramoto, Hidetoshi; Takahashi, Toru; Niimura, Sueo

    2014-01-01

    Fetal bovine serum (FBS) has been widely used as a supplement in the maturation medium of bovine oocytes in vitro. However, serum contains many undefined factors and is potentially infectious to humans and animals. As a serum replacement, we evaluated the feasibility of using the silk protein, sericin, derived from the cocoons of silkworm. To examine the rates of oocyte maturation and fertilization, cumulus-oocyte complexes were cultured in TCM-199 supplemented with 0.01%, 0.05%, 0.1% or 0.15% sericin or 5% FBS. The sizes of the perivitelline space that might relate to polyspermy, the expressions of Has2 and CD44 mRNA, the amount of hyaluronan (hyaluronic acid: HA) contained in the oocytes and the rates of blastocyst formation following insemination were then compared between the oocytes cultured with 0.05% sericin and 5% FBS, because the polyspermy rates in oocytes cultured with 0.05% sericin were significantly lower than in those cultured with 5% FBS. After in vitro maturation (IVM), the mean size of the perivitelline space was significantly greater in oocytes cultured with sericin than in those cultured with FBS, although the rates of nuclear maturation, fertilization and blastocyst formation of oocytes under both IVM conditions were not significantly different. The expression of HAS2 and CD44 mRNA and the amount of HA in the denuded oocytes cultured with 0.05% sericin were significantly greater than in those cultured with FBS. These results indicate the feasibility of sericin as an alternative protein supplement for IVM in bovine oocytes.

  5. [The analysis of physicians' work: announcing the end of attempts at in vitro fertilization].

    Science.gov (United States)

    Santiago-Delefosse, M; Cahen, F; Coeffin-Driol, C

    2003-01-01

    The purpose of this empirical study is to analyze modalities of announcing the end of attempts at in vitro ferti-lization to women who, for various reasons, were not able to have a child after several trials. What are the problems physicians face when, in the course of their work, they make these announcements? How do they give (or not give) support to these women who have placed so much hope in this technique? These are some of the questions that led the authors to conduct this empirical study within the framework of a clinical and qualitative approach to work psychology. Within this framework, work is conceptualised as a complex activity that involves the subject, both bodily and through his various modes of socialisation. The field of clinical and quali-tative approach to work psychology situations focuses on different ways of expressing distress related to contradictory work demands, as the activity is being performed; it also focuses on those creative processes used by the subject to cope with those internal and external conflicts that hinder task performance. A review of the literature and preliminary observations led us to postulate that the problems physicians are faced with when they announce the end of attempts at in vitro fertilisation (IVF) are linked to several conflicts between work values (that are specific to the medical world) and the recognition of work failure: termination of attempts at IVF. The popu-lation that participated in this research project belongs to a network of private practitioners who work with the in-house team of a Parisian clinic. But the group is not uniform and some physicians perform IVFs more frequently than others. Our qualitative study involved 10 semi-directive interviews of approximately 1 1/2 hours each, which were recorded and transcribed. Initial instructions focused on a concrete description of situations of abandonment of attempts at IVF, in terms of their preparation, development, and the way they are experienced

  6. Trolox and ascorbic acid reduce direct and indirect oxidative stress in the IPEC-J2 cells, an in vitro model for the porcine gastrointestinal tract.

    Science.gov (United States)

    Vergauwen, Hans; Tambuyzer, Bart; Jennes, Karen; Degroote, Jeroen; Wang, Wei; De Smet, Stefaan; Michiels, Joris; Van Ginneken, Chris

    2015-01-01

    Oxidative stress in the small intestinal epithelium is a major cause of barrier malfunction and failure to regenerate. This study presents a functional in vitro model using the porcine small intestinal epithelial cell line IPEC-J2 to examine the effects of oxidative stress and to estimate the antioxidant and regenerative potential of Trolox, ascorbic acid and glutathione monoethyl ester. Hydrogen peroxide and diethyl maleate affected the tight junction (zona occludens-1) distribution, significantly increased intracellular oxidative stress (CM-H2DCFDA) and decreased the monolayer integrity (transepithelial electrical resistance and FD-4 permeability), viability (neutral red) and wound healing capacity (scratch assay). Trolox (2 mM) and 1 mM ascorbic acid pre-treatment significantly reduced intracellular oxidative stress, increased wound healing capacity and reduced FD-4 permeability in oxidatively stressed IPEC-J2 cell monolayers. All antioxidant pre-treatments increased transepithelial electrical resistance and viability only in diethyl maleate-treated cells. Glutathione monoethyl ester (10 mM) pre-treatment significantly decreased intracellular oxidative stress and monolayer permeability only in diethyl maleate-treated cells. These data demonstrate that the IPEC-J2 oxidative stress model is a valuable tool to screen antioxidants before validation in piglets.

  7. The in vitro protection of human decay accelerating factor and hDAF/heme oxygenase-1 transgenes in porcine aortic endothelial cells against sera of Formosan macaques.

    Science.gov (United States)

    Tu, C-F; Tai, H-C; Wu, C-P; Ho, L-L; Lin, Y-J; Hwang, C-S; Yang, T-S; Lee, J-M; Tseng, Y-L; Huang, C-C; Weng, C-N; Lee, P-H

    2010-01-01

    To mitigate hyperacute rejection, pigs have been generated with alpha-Gal transferase gene knockout and transgenic expression of human decay accelerating factor (hDAF), MCP, and CD59. Additionally, heme-oxygenase-1 (HO-1) has been suggested to defend endothelial cells. Sera (MS) (0%, 1%, 5%, 10%, and 15%) from Formosan macaques (Macaca cyclopis, MC), an Old World monkey wildly populated in Taiwan, was used to test the protective in vitro, effects of hDAF or hDAF/hHO-1 on porcine aortic endothelial cells (pAEC) derived from hDAF(+), hDAF(+)/hHO-1(+), and hDAF(+)/hHO-1(-) and 1 nontransgenic pAEC. Ten percent human serum (HS) served as a positive control. When MS addition increased to 10% or 15%, all transgenic pAEC exhibited a greater survival than nontransgenic pAEC. Noticeably, 15% MS reduced survived to 40% in nontransgenic and transgenic pAEC, respectively. These results revealed that hDAF exerted protective effects against MC complement activation. However, comparing with 10% MS and HS in pAEC of nontransgenic pigs, the survivability was higher in HS, suggesting that complement activation by MS was more toxic than that by HS. Furthermore, hDAF(+)/hHO-1(+) showed no further protection against effects of MS on transgenic pAEC. Copyright 2010 Elsevier Inc. All rights reserved.

  8. Purification and characterization of porcine liver DNA polymerase gamma: utilization of dUTP and dTTP during in vitro DNA synthesis.

    Science.gov (United States)

    Mosbaugh, D W

    1988-06-24

    Porcine liver DNA polymerase gamma has been demonstrated to preferentially incorporate dTMP over dUMP during in vitro DNA synthesis. When polymerase activity was measured in standard reactions containing saturating levels of either dTTP or dUTP, the polymerization rate was slightly faster in the reaction containing dTTP. However, under conditions where both dTTP and dUTP competed, at an equal molar concentration, approximately 3-times more thymine residues were incorporated than uracil residues into DNA. Similarly, preferential incorporation of dTMP was observed on several substrates including poly (dA).oligo p(dT), poly (rA).oligo p(dT) and poly (dA-dT). The discrimination against dUMP incorporation was even more apparent with reduced levels of dUTP. These observations were consistent with the finding that the Km for DNA polymerase gamma was about 3-fold lower for dTTP (0.4 microM) than for dUTP (1.1 microM). On the other hand, the Vmax for these two reactions was very similar. Discrimination against dUMP incorporation was also observed during inhibition of polymerase gamma by dideoxyribonucleoside triphosphates. Dideoxythymidine triphosphate preferentially inhibited dUMP incorporation compared to that of dTMP, whereas ddATP, ddCTP and ddGTP inhibited both reactions equally.

  9. A comparison of perinatal outcomes in singletons and multiples born after in vitro fertilization or intracytoplasmic sperm injection stratified for neonatal risk criteria

    NARCIS (Netherlands)

    van Heesch, M.M.J.; Evers, J.L.H.; Dumoulin, J.C.M.; van der Hoeven, M.A.H.B.; van Beijsterveldt, C.E.M.; Bonsel, G.J.; Dykgraaf, R.H.M.; van Goudoever, J.B.; Koopman-Esseboom, C.; Nelen, W.L.D.M.; Steiner, K.; Tamminga, P.; Tonch, N.; van Zonneveld, P.; Dirksen, C.D.

    2014-01-01

    Objective To compare perinatal singleton and multiple outcomes in a large Dutch in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) population and within risk subgroups. Newborns were assigned to a risk category based on gestational age, birthweight, Apgar score and congenital

  10. A comparison of perinatal outcomes in singletons and multiples born after in vitro fertilization or intracytoplasmic sperm injection stratified for neonatal risk criteria

    NARCIS (Netherlands)

    Heesch, M.M. van; Evers, J.L.H.; Dumoulin, J.C.; Hoeven, M.A. van der; Beijsterveldt, C.E. van; Bonsel, G.J.; Dykgraaf, R.H.; Goudoever, J.B. van; Koopman-Esseboom, C.; Nelen, W.L.D.M.; Steiner, K.; Tamminga, P.; Tonch, N.; Zonneveld, P. van; Dirksen, C.D.

    2014-01-01

    OBJECTIVE: To compare perinatal singleton and multiple outcomes in a large Dutch in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) population and within risk subgroups. Newborns were assigned to a risk category based on gestational age, birthweight, Apgar score and congenital

  11. Estudo comparativo in vitro entre biopróteses de pericárdio bovino e porcinas In vitro comparative study between bovine pericardial and porcine bioprosthesis

    Directory of Open Access Journals (Sweden)

    Domingo M Braile

    1996-12-01

    Full Text Available A maioria dos implantes valvulares cardíacos realizados no Brasil é representada pelas válvulas de pericárdio bovino, seguidas por próteses porcinas. Na avaliação de válvulas biológicas, deve-se considerar: desempenho hidrodinámico, resistência à fadiga e processo de calcificação. No presente estudo, foi avaliado o desempenho hidrodinámico de biopróteses de pericárdio bovino (Biopro-PB-Braile Biomédica comparativamente às válvulas porcinas (Biopro-PP-Braile Biomédica através do gradiente médio transvalvular. Os testes hidrodinámicos foram realizados em próteses de diâmetros variando de 19 a 35 mm, submetidas ao Sistema Duplicador de Pulsos Shelhigh (Shelhigh Inc.. O volume de ejeção foi mantido constante em 90 ml, com freqüência de pulso de 60, 70,80, 90 e 100 ciclos por minuto, possibilitando fluxos entre 5 e 9 litros por minuto, equivalentes a fluxos contínuos aproximados de 8 a 18 litros por minuto. Houve tendência à diminuição dos gradientes pressóricos à medida em que aumenta o diâmetro externo das próteses. O gradiente pressórico médio encontrado em próteses de pericárdio bovino foi significativamente menor que o de próteses porcinas (pMost of the cardiac valve implantations in Brazil are represented by bovine pericardial valves, followed by the porcine prostheses. In the evaluation of biological valves, the following should be taken into consideration: hydrodynamic performance resistance to fatigue and calcification process. In this study, the hydrodynamic performance of bovine pericardial bioprostheses (Biopro-BP-Braile Biomedica was evaluated comparatively to porcine valves (Biopro-PP-Braile Biomedica, through the transvalvular medium gradient. The hydrodynamic tests were made on prostheses varying from 19 to 35 mm in diameter, which underwent the pulse duplicator system Shelhigh (Shelligh Inc.. The ejection volume was constantly kept at 90 ml. The pulse frequencies varied between 60 and 100

  12. Attenuation of virulence of Lawsonia intracellularis after in vitro passages and its effects on the experimental reproduction of porcine proliferative enteropathy.

    Science.gov (United States)

    Vannucci, Fabio A; Beckler, Dana; Pusterla, Nicola; Mapes, Samantha M; Gebhart, Connie J

    2013-02-22

    Non-pathogenic Lawsonia intracellularis variants have been obtained through multiple passages in cell culture but there is no information regarding the number of passages necessary to attenuate a pathogenic isolate. The present study evaluated the susceptibility of pigs to L. intracellularis after 10, 20 and 40 passages in vitro. Three groups (six animals/group) were inoculated with pure culture of L. intracellularis on passage 10, 20 or 40 and one group with placebo. The animals were monitored for clinical signs, fecal shedding and serological IgG response during 28 days post-inoculation. Gross and histologic lesions and the level of infection based on the amount of L. intracellularis-specific antigen in the intestinal mucosa identified by immunohistochemistry were evaluated in two animals from each group on days 14, 21 and 28. Animals inoculated with passages 10 and 20 demonstrated proliferative lesions typical of porcine proliferative enteropathy associated with the presence of Lawsonia-specific antigen in the intestinal mucosa. Passage 40-inoculated pigs did not show proliferative lesions or presence of Lawsonia antigen at any time point throughout the study. Similar patterns of the fecal shedding were observed in passage 10 and 20-infected pigs but those infected with passage 40 shed for a short period. Serological IgG responses in passage 10 and 20-inoculated pigs were detected from day 14 post-infection but not at all in passage 40-inoculated animals. These results demonstrate attenuation of the virulence properties of L. intracellularis between 20 and 40 cell passages in vitro. This information will be valuable for design of future experimental models and for studying the mechanisms involved in the attenuation of L. intracellularis virulence. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Adapted preparation technique for screw-type implants: explorative in vitro pilot study in a porcine bone model.

    Science.gov (United States)

    Beer, Andreas; Gahleitner, André; Holm, Anders; Birkfellner, Wolfgang; Homolka, Peter

    2007-02-01

    The aim of this study was to quantify the effect of adapted preparation on the insertion torque of self-tapping implants in cancellous bone. In adapted preparation, bone condensation - and thus, insertion torque - is controlled by changing the diameter of the drilling. After preparation of cancellous porcine vertebral bone with drills of 2.85, 3, 3.15 or 3.35 mm final diameters, Brånemark sytem Mk III implants (3.75 x 11.5 mm) were inserted in 141 sites. During implantation, the insertion torque was recorded. Prior to implant insertion, bone mineralization (bone mineral density (BMD)) was measured with dental quantative computed tomography. The BMD values measured at the implant position were correlated with insertion torque for varying bone condensation. Based on the average torque recorded during implant insertion into the pre-drilled canals with a diameter of 3 mm, torque increased by approximately 17% on reducing the diameter of the drill by 5% (to 2.85 mm). On increasing the diameter of the osteotomy to 3.15 mm (5%) or 3.35 mm (12%), torque values decreased by approximately 21% and 50%, respectively. The results demonstrate a correlation between primary stability (average insertion torque) and the diameter of the implant bed on using a screw-shaped implant. Thus, using an individualized bone mineralization-dependent drilling technique, optimized torque values could be achieved in all tested bone qualities with BMDs ranging from 330 to 500 mg/cm(3). The results indicate that using a bone-dependent drilling technique, higher torque values can also be achieved in poor bone using an individualized drilling resulting in higher bone condensation. As immediate function is dependent on primary stability (high insertion torque), this indicates that primary stability can be increased using a modified drilling technique in lesser mineralized bone.

  14. Prediction of formulation effects on dermal absorption of topically applied ectoparasiticides dosed in vitro on canine and porcine skin using a mixture-adjusted quantitative structure permeability relationship.

    Science.gov (United States)

    Riviere, J E; Brooks, J D; Collard, W T; Deng, J; de Rose, G; Mahabir, S P; Merritt, D A; Marchiondo, A A

    2014-10-01

    Topical application of ectoparasiticides for flea and tick control is a major focus for product development in animal health. The objective of this work was to develop a quantitative structure permeability relationship (QSPeR) model sensitive to formulation effects for predicting absorption and skin deposition of five topically applied drugs administered in six vehicle combinations to porcine and canine skin in vitro. Saturated solutions (20 μL) of (14) C-labeled demiditraz, fipronil, permethrin, imidacloprid, or sisapronil were administered in single or binary (50:50 v/v) combinations of water, ethanol, and transcutol (6 formulations, n = 4-5 replicates per treatment) nonoccluded to 0.64 cm(2) disks of dermatomed pig or dog skin mounted in flow-through diffusion cells. Perfusate flux over 24 h and skin deposition at termination were determined. Permeability (logKp), absorption, and penetration endpoints were modeled using a four-term Abrahams and Martin (hydrogen-bond donor acidity and basicity, dipolarity/polarizability, and excess molar refractivity) linear free energy QSPeR equation with a mixture factor added to compensate for formulation ingredient interactions. Goodness of fit was judged by r(2) , cross-validation coefficient, coefficients (q(2) s), and Williams Plot to visualize the applicability domain. Formulation composition was the primary determinant of permeation. Compounds generally penetrated dog skin better than porcine skin. The vast majority of permeated penetrant was deposited within the dosed skin relative to transdermal flux, an attribute for ectoparasiticides. The best QSPeR logKp model for pig skin permeation (r(2) = 0.86, q(2) s = 0.85) included log octanol/water partition coefficient as the mixture factor, while for dogs (r(2) = 0.91, q(2) s = 0.90), it was log water solubility. These studies clearly showed that the permeation of topical ectoparasiticides could be well predicted using QSPeR models that account for both the physical

  15. Primary cilia on porcine testicular somatic cells and their role in hedgehog signaling and tubular morphogenesis in vitro.

    Science.gov (United States)

    Dores, Camila; Alpaugh, Whitney; Su, Lin; Biernaskie, Jeff; Dobrinski, Ina

    2017-04-01

    The primary cilium is a microtubule-based sensory organelle found on nearly all eukaryotic cells but little is understood about its function in the testis. We investigate the role of primary cilia on testis cells in vitro by inhibiting formation of the primary cilium with Ciliobrevin D, a cell-permeable, reversible chemical inhibitor of ATPase motor cytoplasmic dynein. We analyzed cultured cells for the presence of primary cilia and their involvement in hedgehog signaling. Primary cilia were present on 89.3 ± 2.3 % of untreated testicular somatic cells compared to 3.1 ± 2.5 % cells with primary cilia for Ciliobrevin D-treated cells. Protein levels of Gli-2 and Smoothened were lower on Western blots after suppression of cilia with Ciliobrevin D. The inhibitor did not affect centrosome localization or cell proliferation, indicating that changes were due to ablation of the primary cilium. Testicular somatic cells have the ability to form three-dimensional tubules in vitro. In vitro-formed tubules were significantly longer and wider in the control group than in the Ciliobrevin D-treated group (9.91 ± 0.35 vs. 5.540 ± 1.08 mm and 339.8 ± 55.78 vs. 127.2 ± 11.9 μm, respectively) indicating that primary cilia play a role in tubule formation. Our results establish that the inhibition of ATPase motor cytoplasmic dynein perturbs formation of primary cilia in testicular somatic cells, affects the hedgehog signaling pathway and impairs tubule formation in vitro. These findings provide evidence for a role of cilia in the testis in cell signaling and tubular morphogenesis in vitro.

  16. D-penicillamine and granulosa cells can effectively extend the fertile life span of bovine frozen-thawed spermatozoa in vitro: effect on fertilization and polyspermy.

    Science.gov (United States)

    Pavlok, A

    2000-03-15

    The effect of D-penicillamine on the fertile life span of frozen-thawed bull spermatozoa was studied in Experiment 1. After thawing, the washed spermatozoa were incubated for 8 h in fertilization medium with 1 mg/mL polyvinyl alcohol (PVA) and 0.5 mg/mL D-penicillamine. The addition of cumulus-free oocytes together with 10% of bovine serum (BOS) + 4 IU/mL heparin to the 8-h incubated spermatozoa resulted in high fertilization and polyspermy rates (97/103, 94.2% and 49/97, 50.5%, respectively). When BOS was substituted with 3 mg/mL of BSA, the fertilization and polyspermy rates decreased to 33/91 (36.3%) and 3/33 (9.1%), respectively. The total absence of fertilization was observed after substitution of proteins with PVA. The 8-h sperm incubation in fertilization medium without D-penicillamine and following fertilization in medium + different capacitation supplements resulted in the total absence of or a very low fertilization rate. In Experiment 2, the first set of cumulus-oocyte complexes (COC) and cumulus-free oocytes were fertilized for 8 h with nonincubated spermatozoa. The second set of COC and cumulus-free oocytes were fertilized in the same wells with spermatozoa after removal of the first set. The fertilization rate for the first set of COC and cumulus-free oocytes was 65/67 (97%) and 73/73 (100%), respectively, with 21/65 (32.3%) and 32/73 (43.8%) polyspermy, respectively. In the second set, the high penetration rate (67/73, 91.8%) was observed only for COC, while that for cumulus-free oocytes (19/76, 25%) was significantly lower (P polyspermy. In medium + BSA + 10 or 100 IU/mL heparin, the fertilization rate of COC was 64/72 (88.9%) and 70/79 (88.6%), respectively, with polyspermy at 2/64 (3.1%) and 7/70 (10%), respectively. In medium + BOS + 10 or 100 IU/mL heparin, the fertilization rate was 79/82 (96.3%) and 79/80 (98.7%), respectively, with a significantly (P polyspermy rate (34/79, 43% and 30/79, 38%, respectively). The vitality and capacitation

  17. A comparative study of saffron aqueous extract and its active ingredient, crocin on the in vitro maturation, in vitro fertilization, and in vitro culture of mouse oocytes.

    Science.gov (United States)

    Mokhber Maleki, Elham; Eimani, Hussein; Bigdeli, Mohammad Reza; Ebrahimi, Bita; Shahverdi, Abdol Hossein; Golkar Narenji, Afsane; Abedi, Reyhane

    2014-03-01

    Reactive oxygen species have effects on gamete quality and gamete interaction; they influence spermatozoa, oocytes, embryos, and their environment. In this study, we evaluated the antioxidant effect of different concentrations of saffron (Crocus sativus L.) aqueous extract (SAE) and its ingredient, crocin, on the improvement of in vitro maturation (IVM) and subsequent in vitro fertilization (IVF) and embryo development of mouse oocytes. Cumulus oocyte complexes were collected from ovaries, and germinal vesicle oocytes were cultured in the presence of SAE and crocin. SAE was added at dosages of 5 μg/mL, 10 μg/mL, and 40 μg/mL; dosages of crocin were 50 μg/mL, 100 μg/mL, and 400 μg/mL. All dosages were added to maturation medium and a group without SAE or crocin was considered as the control group. Following IVM, metaphase II oocytes were fertilized and cultured in vitro in order to observe embryo development. Both SAE and crocin improved the rate of IVM, IVF, and in vitro culture. Addition of 40 μg/mL SAE to maturation medium significantly increased the rate of IVM, IVF, and in vitro culture (p < 0.05). Furthermore 100 μg/mL crocin significantly increased the IVM rate compared to the control group (p < 0.05). Use of SAE during IVM can affect on IVM, IVF, and early embryo development in a dose-dependent manner. SAE appears to have a stronger effect than pure crocin. Copyright © 2014. Published by Elsevier B.V.

  18. Ultrasound-mediated transdermal drug delivery of fluorescent nanoparticles and hyaluronic acid into porcine skin in vitro

    Science.gov (United States)

    Wang, Huan-Lei; Fan, Peng-Fei; Guo, Xia-Sheng; Tu, Juan; Ma, Yong; Zhang, Dong

    2016-12-01

    Transdermal drug delivery (TDD) can effectively bypass the first-pass effect. In this paper, ultrasound-facilitated TDD on fresh porcine skin was studied under various acoustic parameters, including frequency, amplitude, and exposure time. The delivery of yellow-green fluorescent nanoparticles and high molecular weight hyaluronic acid (HA) in the skin samples was observed by laser confocal microscopy and ultraviolet spectrometry, respectively. The results showed that, with the application of ultrasound exposures, the permeability of the skin to these markers (e.g., their penetration depth and concentration) could be raised above its passive diffusion permeability. Moreover, ultrasound-facilitated TDD was also tested with/without the presence of ultrasound contrast agents (UCAs). When the ultrasound was applied without UCAs, low ultrasound frequency will give a better drug delivery effect than high frequency, but the penetration depth was less likely to exceed 200 μm. However, with the help of the ultrasound-induced microbubble cavitation effect, both the penetration depth and concentration in the skin were significantly enhanced even more. The best ultrasound-facilitated TDD could be achieved with a drug penetration depth of over 600 μm, and the penetration concentrations of fluorescent nanoparticles and HA increased up to about 4-5 folds. In order to get better understanding of ultrasound-facilitated TDD, scanning electron microscopy was used to examine the surface morphology of skin samples, which showed that the skin structure changed greatly under the treatment of ultrasound and UCA. The present work suggests that, for TDD applications (e.g., nanoparticle drug carriers, transdermal patches and cosmetics), protocols and methods presented in this paper are potentially useful. Project partially supported by the National Natural Science Foundation of China (Grant Nos. 81127901, 81227004, 81473692, 81673995, 11374155, 11574156, 11274170, 11274176, 11474001

  19. Effect of caffeine on motility and vitality of sperm and in vitro fertilization of outbreed mouse in T6 and M16 media.

    Science.gov (United States)

    Nabavi, Narges; Todehdehghan, Fatemeh; Shiravi, Abdollhossein

    2013-09-01

    Caffeine increases the CAMP production that stimulates spermatozoa movement. Caffeine is also used for induction of in vitro acrosome reaction in mammalian spermatozoa, an important step in achieving fertilization. The aim of this study was to assess the effect of caffeine on sperm's motility, vitality and laboratory fertilization rates in mouse in two T6 and M16 media. Epididymal mouse sperms were collected and treated by caffine in T6 and M16 media and their motility and vitality rates were evaluated. The pretreated sperms were added to oocytes in T6 and M16 media with and without caffeine and fertilization rates were recorded after 24 hours incubation. Sperm's motility (81.7±1.67%) and vitality (88.7±1.33%) rates and percentage of fertilized oocytes (67.52±8.16%) in T6 medium plus caffeine compare to control group have increased and shown significant differences at p≤0.01. While the percentages of these parameters in M16 medium supplemented with caffeine were 68.3±6.01%, 78±6.11%, and 42.6±12.96 respectively and in comparison to control group (M16 without caffeine) have not shown significant differences. Addition of caffeine to T6 medium promotes the sperm's motility and vitality and enhances fertilization and early in vitro development of mouse embryos. This article extracted from M.Sc. thesis. (Narges Navabi).

  20. Technique of the `in vitro` fertilization and the culture of mouse embryos at preimplantation; Tecnica de fertilizacao `in vitro` e cultura de embrioes de camundongo durante a pre-implantacao

    Energy Technology Data Exchange (ETDEWEB)

    Kikuchi, Olivia Kimiko [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil); Yamada, Takeshi [National Inst. of Radiological Sciences, Chiba (Japan)

    1993-03-01

    The mammal embryo is an intensive cellular proliferating system, very radiosensitive and therefore adequate to the study of the biological effects of ionizing radiation. The technique of the in vitro fertilization and the culture of mouse embryos at preimplantation period, modified by Yamada et al (1982) to improve the efficiency of more than 95% of blastocyst formation is described. (author) 2 refs., 7 figs.

  1. In Vitro Fertilization in Women With Inflammatory Bowel Disease Is as Successful as in Women From the General Infertility Population

    Science.gov (United States)

    Oza, Sveta Shah; Pabby, Vikas; Dodge, Laura E.; Moragianni, Vasiliki A.; Hacker, Michele R.; Fox, Janis H.; Correia, Katharine; Missmer, Stacey A.; Ibrahim, Yetunde; Penzias, Alan S.; Burakoff, Robert; Friedman, Sonia; Cheifetz, Adam S.

    2015-01-01

    BACKGROUND & AIMS Inflammatory bowel disease (IBD) affects women of reproductive age, so there are concerns about its effects on fertility. We investigated the success of in vitro fertilization (IVF) in patients with IBD compared with the general (non-IBD) IVF population. METHODS We conducted a matched retrospective cohort study of female patients with IBD who under-went IVF from 1998 through 2011 at 2 tertiary care centers. Patients were matched 4:1 to those without IBD (controls). The primary outcome was the cumulative rate of live births after up to 6 cycles of IVF. Secondary outcomes included the proportion of patients who became pregnant and the rate of live births for each cycle. RESULTS Forty-nine patients with Crohn’s disease (CD), 71 patients with ulcerative colitis (UC), 1 patient with IBD-unclassified, and 470 controls underwent IVF during the study period. The cumulative rate of live births was 53% for controls, 69% for patients with UC (P = .08 compared with controls), and 57% for patients with CD (P = .87 compared with controls). The incidence of pregnancy after the first cycle of IVF was similar among controls (40.9%), patients with UC (49.3%; P = .18), and patients with CD (42.9%; P = .79). Similarly, the incidence of live births after the first cycle of IVF was similar among controls (30.2%), patients with UC (33.8%; P = .54), and patients with CD (30.6%; P = .95). CONCLUSIONS Based on a matched cohort study, infertile women with IBD achieve rates of live births after IVF that are comparable with those of infertile women without IBD. PMID:25818081

  2. In vitro fertilization pregnancy rates in levothyroxine-treated women with hypothyroidism compared to women without thyroid dysfunction disorders.

    Science.gov (United States)

    Scoccia, Bert; Demir, Habibe; Kang, Yuna; Fierro, Michelle A; Winston, Nicola J

    2012-06-01

    Untreated hypothyroidism can lead to ovulatory dysfunction resulting in oligo-amenorrhea. Treatment with levothyroxine can reverse such dysfunction and thus should improve fertility. The purpose of this retrospective study was to assess whether in vitro fertilization (IVF) pregnancy rates differ in levothyroxine-treated women with hypothyroidism compared to women without thyroid dysfunction/disorders. Treated hypothyroid and euthyroid women undergoing IVF at an academic IVF center were studied after Institutional Review Board approval. Women with hypothyroidism were treated with levothyroxine 0.025-0.15 mg/day for at least 3 months to maintain baseline thyrotropin (TSH) levels of 0.35-4.0 μU/mL prior to commencing IVF treatment (HYPO-Rx group). Causes of infertility were similar in both groups with the exception of male factor, which was more common in the HYPO-Rx group. The main outcomes studied were implantation rate, clinical pregnancy rate, clinical miscarriage rate, and live birth rate. We reviewed the first IVF retrieval cycle performed on 240 women aged 37 years or less during the period January 2003 to December 2007. Women with treated hypothyroidism (n=21) had significantly less implantation, clinical pregnancy, and live birth rates than euthyroid women (n=219). We conclude that, despite levothyroxine treatment, women with hypothyroidism have a significantly decreased chance of achieving a pregnancy following IVF compared to euthyroid patients. A larger prospective study is necessary to assess confounding variables, confirm these findings, and determine the optimal level of TSH prior to and during controlled ovarian hyperstimulation for IVF.

  3. In Vitro Fertilization Pregnancy Rates in Levothyroxine-Treated Women With Hypothyroidism Compared to Women Without Thyroid Dysfunction Disorders

    Science.gov (United States)

    Demir, Habibe; Kang, Yuna; Fierro, Michelle A.; Winston, Nicola J.

    2012-01-01

    Background Untreated hypothyroidism can lead to ovulatory dysfunction resulting in oligo-amenorrhea. Treatment with levothyroxine can reverse such dysfunction and thus should improve fertility. The purpose of this retrospective study was to assess whether in vitro fertilization (IVF) pregnancy rates differ in levothyroxine-treated women with hypothyroidism compared to women without thyroid dysfunction/disorders. Methods Treated hypothyroid and euthyroid women undergoing IVF at an academic IVF center were studied after Institutional Review Board approval. Women with hypothyroidism were treated with levothyroxine 0.025–0.15 mg/day for at least 3 months to maintain baseline thyrotropin (TSH) levels of 0.35–4.0 μU/mL prior to commencing IVF treatment (HYPO-Rx group). Causes of infertility were similar in both groups with the exception of male factor, which was more common in the HYPO-Rx group. The main outcomes studied were implantation rate, clinical pregnancy rate, clinical miscarriage rate, and live birth rate. Results We reviewed the first IVF retrieval cycle performed on 240 women aged 37 years or less during the period January 2003 to December 2007. Women with treated hypothyroidism (n=21) had significantly less implantation, clinical pregnancy, and live birth rates than euthyroid women (n=219). Conclusions We conclude that, despite levothyroxine treatment, women with hypothyroidism have a significantly decreased chance of achieving a pregnancy following IVF compared to euthyroid patients. A larger prospective study is necessary to assess confounding variables, confirm these findings, and determine the optimal level of TSH prior to and during controlled ovarian hyperstimulation for IVF. PMID:22540326

  4. A Comprehensive Analysis of Body Mass Index Effect on in Vitro Fertilization Outcomes

    Directory of Open Access Journals (Sweden)

    Veronica Sarais

    2016-02-01

    Full Text Available The effect of a raised body mass index (BMI on the outcome of assisted reproduction technology (ART still represents a controversial issue. Even less clear is whether BMI acts with a potential detrimental effect on IVF outcomes via a deleterious effect on innate quality of oocytes or on the environmental milieu within the uterus. With the aim to better understand the mechanisms underlying the potential deleterious effect of an increased BMI on IVF outcomes, we have evaluated the effects of female BMI on number and quality of retrieved oocytes, fertilization rate, embryo score and incidences of ongoing pregnancy and live births among couples undergoing IVF in an Italian population. Data from 1602 women who underwent their first IVF cycle were retrospectively analyzed. A significantly reduced percentage of mature oocytes when comparing obese (BMI ≥ 30 kg/m2 and normal-weight patients (BMI = 18.50–24.99 kg/m2 was found. After adjusting for maternal age and other confounders, odds for ongoing pregnancy rate showed no differences across different BMI categories. However, a significant increased odds ratio (OR could be observed for miscarriage rate in patients with BMI ≥ 25 (OR = 2.5; p = 0.04. These results should be taken into account in order to define optimal strategies for overweight and obese patients referring to ART procedures.

  5. The role of gonadotropin-releasing hormone antagonists in in vitro fertilization.

    Science.gov (United States)

    Diedrich, K; Ludwig, M; Felberbaum, R E

    2001-09-01

    Gonadotropin-releasing hormone (GnRH)-antagonists can suppress the pituitary hormone secretion completely within a few hours, allowing the avoidance of premature luteinization within controlled ovarian hyperstimulation (COH) for assisted reproductive technologies (ART) by midcycle administration. Two different protocols were described, which were widely used in COH in several phase II and III studies as well as in clinical practice since the GnRH-antagonists Cetrorelix (Cetrotidesound recording copyright sign; Serono International S.A., Geneva, Switzerland) and Ganirelix (Orgalutansound recording copyright sign, Antagonsound recording copyright sign; Organon, Oss, The Netherlands) are available on the market. Cetrorelix was applied in single- and multiple-dose protocols; Ganirelix was used until now only according to the multiple-dose protocol. Fertilization rates of >60% as well as clinical pregnancy rates of about 30% per transfer sound most promising. Estradiol secretion is not compromised by the GnRH-antagonists using recombinant follicle-stimulating hormone (FSH) for COH. The incidence of a premature leutinizing hormone (LH) surge is far below 2% while the pituitary response remains preserved, allowing the induction of ovulation by GnRH or GnRH-agonists. However, luteal phase support remains mandatory. The incidence of severe ovarian hyperstimulation syndrome (OHSS) seems to be lower under antagonist treatment than in the long agonistic protocol. Treatment time is significantly shortened. Without any doubt GnRH-antagonists have the potential to become the new standard for controlled ovarian hyperstimulation.

  6. Fertilization in vitro with spermatozoa from different mice increased variation in the developmental potential of embryos compared to artificial parthenogenetic activation.

    Science.gov (United States)

    Miao, De-Qiang; Liang, Bo; Wang, Jun-Zuo; Wang, Hui-Li; Cui, Wei; Liu, Yong; Tan, Jing-He

    2009-03-01

    Although successful embryo development is dependent upon genetic and epigenetic contributions from both the male and female, the male potential to adversely affect embryo development has been scarcely studied. It is unclear whether the sperm variation among different males would affect the outcome of oocyte evaluation by embryo development following fertilization. In the present study, variation in the developmental potential of mouse embryos was first compared between in vitro fertilization with epididymal spermatozoa from different males and Sr(2+) parthenogenetic activation using oocytes of different qualities, and then the effect of male on fertilization and embryo development was examined using randomly chosen oocytes and spermatozoa from cauda epididymidis, vas deferens or electro-ejaculates. Rates of fertilization and blastocyst formation were significantly higher with spermatozoa from cauda epididymidis or vas deferens than with ejaculated spermatozoa. Rates of embryonic development differed significantly between different males, but not between different ejaculates of the same male. Analysis of standard errors of means and coefficients of variance indicated that as long as multiple males were involved, the variation in oocyte fertilization/activation and blastocyst formation was always higher after fertilization than after Sr(2+) parthenogenetic activation whether spermatozoa were collected from epididymidis, vas deferens or ejaculates and regardless of oocyte qualities. It is concluded that (1) epididymal mouse spermatozoa fertilize more oocytes than ejaculated spermatozoa under identical experimental conditions; (2) like farm animals, the mice also show a remarkable male effect on the developmental potential of in vitro produced embryos although they are supposed to be less genetically diverse; (3) parthenogenetic activation is recommended for assessment of oocyte quality to exclude the effect of male.

  7. In vitro fertilization and preterm delivery, low birth weight, and admission to the neonatal intensive care unit: a prospective follow-up study.

    Science.gov (United States)

    Wisborg, Kirsten; Ingerslev, Hans Jacob; Henriksen, Tine Brink

    2010-11-01

    To compare the risk of preterm delivery, low birth weight, and admission of the newborn to a neonatal intensive care unit (NICU) in women pregnant after fertility treatment and subfertile women with the risk in fertile women. Prospective follow-up study. Aarhus University Hospital, Skejby, Denmark, 1989-2006. A total of 20,080 liveborn singletons. None. Preterm delivery, low birth weight, and admission of the newborn to a NICU. After adjustment we found a statistically significantly increased risk of preterm delivery and very preterm delivery in women who conceived after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) compared with fertile women. Compared with fertile women, the risk of preterm delivery and very preterm delivery was not statistically significantly different in women pregnant after non-IVF assisted reproductive treatment (non-IVF ART) or subfertile women. We found no association between IVF/ICSI and the risk of low birth weight at term or admittance to the NICU. The increased risk of preterm delivery after IVF/ICSI may be due to the fertility treatment or unknown characteristics in the couples who undergo IVF/ICSI. Copyright © 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. Urinary Phthalate Metabolite Concentrations and Reproductive Outcomes among Women Undergoing in Vitro Fertilization: Results from the EARTH Study.

    Science.gov (United States)

    Hauser, Russ; Gaskins, Audrey J; Souter, Irene; Smith, Kristen W; Dodge, Laura E; Ehrlich, Shelley; Meeker, John D; Calafat, Antonia M; Williams, Paige L

    2016-06-01

    Evidence from both animal and human studies suggests that exposure to phthalates may be associated with adverse female reproductive outcomes. We evaluated the associations between urinary concentrations of phthalate metabolites and outcomes of assisted reproductive technologies (ART). This analysis included 256 women enrolled in the Environment and Reproductive Health (EARTH) prospective cohort study (2004-2012) who provided one to two urine samples per cycle before oocyte retrieval. We measured 11 urinary phthalate metabolites [mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-oxohexyl) phthalate (MEOHP), mono(2-ethyl-5-carboxypentyl) phthalate (MECPP), mono-isobutyl phthalate (MiBP), mono-n-butyl phthalate (MBP), monobenzyl phthalate (MBzP), monoethyl phthalate (MEP), monocarboxyisooctyl phthalate (MCOP), monocarboxyisononyl phthalate (MCNP), and mono(3-carboxypropyl) phthalate (MCPP)]. We used generalized linear mixed models to evaluate the association of urinary phthalate metabolites with in vitro fertilization (IVF) outcomes, accounting for multiple IVF cycles per woman. In multivariate models, women in the highest as compared with lowest quartile of MEHP, MEHHP, MEOHP, MECPP, ΣDEHP (MEHP + MEHHP + MEOHP + MECPP), and MCNP had lower oocyte yield. Similarly, the number of mature (MII) oocytes retrieved was lower in the highest versus lowest quartile for these same phthalate metabolites. The adjusted differences (95% CI) in proportion of cycles resulting in clinical pregnancy and live birth between women in the fourth versus first quartile of ΣDEHP were -0.19 (-0.29, -0.08) and -0.19 (-0.28, -0.08), respectively, and there was also a lower proportion of cycles resulting in clinical pregnancy and live birth for individual DEHP metabolites. Urinary concentrations of DEHP metabolites were inversely associated with oocyte yield, clinical pregnancy, and live birth following ART. Hauser R, Gaskins AJ, Souter I, Smith

  9. Systematic in vitro and in vivo characterization of Leukemia-inhibiting factor- and Fibroblast growth factor-derived porcine induced pluripotent stem cells

    DEFF Research Database (Denmark)

    Secher, Jan Ole Bertelsen; Ceylan, Ahmet; Mazzoni, Gianluca

    2017-01-01

    Derivation and stable maintenance of porcine induced pluripotent stem cells (piPSCs) is challenging. We herein systematically analyzed two piPSC lines, derived by lentiviral transduction and cultured under either leukemia inhibitory factor (LIF) or fibroblast growth factor (FGF) conditions, to shed......, only their prescence within the embryonic membranes could be detected. Whole transcriptome analysis of the piPSCs and porcine neonatal fibroblasts showed that they clustered together, but apart from the two pluripotent cell populations of early porcine embryos, indicating incomplete reprogramming....... Indeed, bioinformatic analysis of the pluripotency-related gene network of the LIF- versus FGF-derived piPSCs revealed that ZFP42 (REX1) expression was absent in both piPSC-like cells, whereas it was expressed in the porcine inner cell mass at Day 7/8. A second striking difference was the expression...

  10. Towards establishing an occupational threshold for cumulative shear force in the vertebral joint - an in vitro evaluation of a risk factor for spondylolytic fractures using porcine specimens.

    Science.gov (United States)

    Howarth, Samuel J; Callaghan, Jack P

    2013-03-01

    Injury models for spondylolytic fracture of the pars interarticularis have long considered repetitive shear loading as a risk factor without quantifying the relationship between shear force magnitude and fatigue life. This investigation sought to quantify the relationship using a basic in vitro approach. Thirty-two (16 C3-C4, 16 C5-C6) porcine cervical specimens were exposed to repetitive shear loading to 20%, 40%, 60%, or 80% of their calculated ultimate anterior shear failure tolerance. Shear force was cyclically applied at 1Hz for 21,600cycles or until bone failure was detected. Cumulative shear force and the number of cycles sustained until failure were calculated. Failure patterns were also documented. Cumulative shear and the number of cycles sustained prior to failure demonstrated a strong non-linearly decreasing relationship with increased force magnitude. In particular, sustained cumulative shear by the 40% group was 2.52 and 2.63MN∗s higher than for the 60% and 80% groups (P<0.0001). Despite undergoing an average of 230 more loading cycles, cumulative shear force sustained by the 60% group was not statistically different from the 80% group. Bilateral fractures of the cranial vertebra's pars interarticularis were most common, but less consistent at higher force magnitudes. Our investigation suggested that pars interarticularis damage may begin non-linearly accumulating with shear forces between 20% and 40% of failure tolerance (approximately 430 to 860N). Models of pars interarticularis injury and estimates of cumulative shear exposure may be enhanced from a tissue-based weighting method for low-back shear. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

  11. Transcription Adaptation during In Vitro Adipogenesis and Osteogenesis of Porcine Mesenchymal Stem Cells: Dynamics of Pathways, Biological Processes, Up-Stream Regulators, and Gene Networks.

    Science.gov (United States)

    Bionaz, Massimo; Monaco, Elisa; Wheeler, Matthew B

    2015-01-01

    The importance of mesenchymal stem cells (MSC) for bone regeneration is growing. Among MSC the bone marrow-derived stem cells (BMSC) are considered the gold standard in tissue engineering and regenerative medicine; however, the adipose-derived stem cells (ASC) have very similar properties and some advantages to be considered a good alternative to BMSC. The molecular mechanisms driving adipogenesis are relatively well-known but mechanisms driving osteogenesis are poorly known, particularly in pig. In the present study we have used transcriptome analysis to unravel pathways and biological functions driving in vitro adipogenesis and osteogenesis in BMSC and ASC. The analysis was performed using the novel Dynamic Impact Approach and functional enrichment analysis. In addition, a k-mean cluster analysis in association with enrichment analysis, networks reconstruction, and transcription factors overlapping analysis were performed in order to uncover the coordination of biological functions underlining differentiations. Analysis indicated a larger and more coordinated transcriptomic adaptation during adipogenesis compared to osteogenesis, with a larger induction of metabolism, particularly lipid synthesis (mostly triglycerides), and a larger use of amino acids for synthesis of feed-forward adipogenic compounds, larger cell signaling, lower cell-to-cell interactions, particularly for the cytoskeleton organization and cell junctions, and lower cell proliferation. The coordination of adipogenesis was mostly driven by Peroxisome Proliferator-activated Receptors together with other known adipogenic transcription factors. Only a few pathways and functions were more induced during osteogenesis compared to adipogenesis and some were more inhibited during osteogenesis, such as cholesterol and protein synthesis. Up-stream transcription factor analysis indicated activation of several lipid-related transcription regulators (e.g., PPARs and CEBPα) during adipogenesis but osteogenesis

  12. Influence of vitamin D levels on in vitro fertilization outcomes in donor-recipient cycles.

    Science.gov (United States)

    Rudick, Briana J; Ingles, Sue Ann; Chung, Karine; Stanczyk, Frank Z; Paulson, Richard J; Bendikson, Kristin A

    2014-02-01

    To elucidate the role of vitamin D in reproduction by examining the relationship between recipient vitamin D levels and pregnancy rates in donor-recipient IVF cycles. Retrospective cohort study. Academic tertiary care center. Ninety-nine recipients of egg donation at University of Southern California Fertility. Serum was collected from egg donor recipients before ET and was tested for vitamin D levels [25(OH)D]. Clinical pregnancy as defined by sonographic presence of a heartbeat at 7-8 weeks of gestation. In a diverse population of 99 recipients (53% Caucasian, 20% Asian, 16% Hispanic, 7% African American), adjusted clinical pregnancy rates were lower among vitamin D-deficient recipients than among vitamin D-replete recipients (37% vs. 78%). Live-birth rates were 31% among vitamin D-deficient recipients, compared with 59% among vitamin D-replete recipients. There were no differences in adjusted clinical pregnancy and live-birth rates among recipients who were vitamin D deficient [25(OH)D<20 ng/mL] vs. among those who were vitamin D insufficient [20 ng/mL ≤ 25(OH)D<30 ng/mL]. Nonreplete vitamin D status [25(OH)D<30 ng/mL] was associated with lower pregnancy rates in recipients of egg donation. Since the oocyte donor-recipient model is able to separate the impact of vitamin D on oocyte vs. endometrium, these data suggest that the effects of vitamin D may be mediated through the endometrium. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  13. Importance of supply integrity for in vitro fertilization and embryo culture.

    Science.gov (United States)

    Morbeck, Dean E

    2012-06-01

    The quality of in vitro culture conditions is a key component of a successful clinical embryology laboratory. Many, but not all, supplies used in the embryology laboratory are screened by the supplier with a bioassay. Embryology laboratories use a variety of approaches to verify the quality of mineral oil, protein, and disposables before clinical use; however, a best practice has not been determined. Some laboratories test every supply, even those already screened by the supplier, whereas other laboratories perform as little testing as possible. Despite screening by the supplier, recent reports of embryo toxicity, specifically with mineral oil, highlight that the integrity of the supply system has gaps. This review describes current bioassay quality control testing and discusses how it applies to screening of products with documented lot-to-lot variation. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  14. Pre-incubation of porcine semen reduces the incidence of polyspermy on embryos derived from low quality oocytes

    Directory of Open Access Journals (Sweden)

    Cláudio Francisco Brogni

    2016-06-01

    Full Text Available ABSTRACT: The main cause of low efficiency of in vitro produced porcine embryos is the high polyspermic penetration rates at fertilization, which is aggravated in low quality oocytes. Experiment 1 evaluated the embryo development in high and low quality oocytes. Experiment 2 evaluated the embryo development and quality of low quality oocytes fertilized with sperm pre-incubated during 0h (control, 0.5h, 1h and 1.5h. Experiment 3 investigated fertilization and monospermic rates of the same groups of Experiment 2. Experiment 4 evaluated embryo development, cell density, fertilization and monospermic rates of high quality oocytes using semen pre incubated during the best time observed in the previous experiments. Cleavage and blastocyst rates were analyzed by chi-square test, and remaining data by ANOVA and Tukey test (P≤0.05. The cleavage (74.8 vs 51.7% and blastocyst (33.7 vs 9.8% rates were greater in oocytes of high versus low quality, with no differences in cell density. Fertilization rates (65.6 to 79.5% were not influenced by pre-incubation time. However, semen pre-incubation during 1.5h increased monospermic penetration (53.3% and cleavage rates (92.5% in low quality oocytes. Blastocyst rate was improved with 1.5h of semen pre incubation; however they were still lower than that observed with high quality control oocytes. Ultimately, pre-incubation did not influence fertilization, monospermic penetration, embryo development rates, nor cell density in oocytes of high quality. Low-quality porcine oocytes resulted in better rates of embryo development if in vitro fertilized with sperm pre-incubated for 1.5 hour.

  15. Susceptibility of human liver cells to porcine endogenous retrovirus.

    Science.gov (United States)

    Lin, Xinzi; Qi, Lin; Li, Zhiguo; Chi, Hao; Lin, Wanjun; Wang, Yan; Jiang, Zesheng; Pan, Mingxin; Gao, Yi

    2013-12-01

    The risk of porcine endogenous retrovirus infection is a major barrier for pig-to-human xenotransplant. Porcine endogenous retrovirus, present in porcine cells, can infect many human and nonhuman primate cells in vitro, but there is no evidence available about in vitro infection of human liver cells. We investigated the susceptibility of different human liver cells to porcine endogenous retrovirus. The supernatant from a porcine kidney cell line was added to human liver cells, including a normal hepatocyte cell line (HL-7702 cells), primary hepatocytes (Phh cells), and a liver stellate cell line (Lx-2 cells), and to human embryonic kidney cells as a reference control. Expression of the porcine endogenous retrovirus antigen p15E in the human cells was evaluated with polymerase chain reaction, reverse transcription-polymerase chain reaction, and Western blot. The porcine endogenous retrovirus antigen p15E was not expressed in any human liver cells (HL-7702, Phh, or Lx-2 cells) that had been exposed to supernatants from porcine kidney cell lines. Porcine endogenous retrovirus-specific fragments were amplified in human kidney cells. Human liver cells tested were not susceptible to infection by porcine endogenous retrovirus. Therefore, not all human cells are susceptible to porcine endogenous retrovirus.

  16. In vitro fertilization and multiple pregnancies: an evidence-based analysis.

    Science.gov (United States)

    2006-01-01

    . IVF was first developed as a method to overcome bilateral Fallopian tube obstruction. The procedure includes several steps: (1) the woman's egg is retrieved from the ovaries; (2) exposed to sperm outside the body and fertilized; (3) the embryo(s) is cultured for 3 to 5 days; and (4) is transferred back to the uterus. IFV is considered to be one of the most effective treatments for infertility today. According to data from the Canadian Assisted Reproductive Technology Registry, the average live birth rate after IVF in Canada is around 30%, but there is considerable variation in the age of the mother and primary cause of infertility. An important advantage of IVF is that it allows for the control of the number of embryos transferred. An elective single embryo transfer in IVF cycles adopted in many European countries was shown to significantly reduce the risk of multiple pregnancies while maintaining acceptable birth rates. However, when number of embryos transferred is not limited, the rate of IVF-associated multiple pregnancies is similar to that of other treatments involving ovarian stimulation. The practice of multiple embryo transfer in IVF is often the result of pressures to increase success rates due to the high costs of the procedure. The average rate of multiple pregnancies resulting from IVF in Canada is currently around 30%. An alternative to IVF is IUI. In spite of reported lower success rates of IUI (pregnancy rates per cycle range from 8.7% to 17.1%) it is generally attempted before IVF due to its lower invasiveness and cost. Two major drawbacks of IUI are that it cannot be used in cases of bilateral tubal obstruction and it does not allow much control over the risk of multiple pregnancies compared with IVF. The rate of multiple pregnancies after IUI with COS is estimated to be about 21% to 29%. Ontario Health Insurance Plan Coverage Currently, the Ontario Health Insurance Plan covers the cost of IVF for women with bilaterally blocked Fallopian tubes only

  17. Obstetric outcomes in women with polycystic ovary syndrome and isolated polycystic ovaries undergoing in vitro fertilization: a retrospective cohort analysis.

    Science.gov (United States)

    Wan, Hei Lok Tiffany; Hui, Pui Wah; Li, Hang Wun Raymond; Ng, Ernest Hung Yu

    2015-03-01

    This retrospective cohort study evaluated the obstetric outcomes in women with polycystic ovary syndrome (PCOS) and isolated polycystic ovaries (PCO) undergoing in vitro fertilization (IVF) treatment. We studied 104 women with PCOS, 184 with PCO and 576 age-matched controls undergoing the first IVF treatment cycle between 2002 and 2009. Obstetric outcomes and complications including gestational diabetes (GDM), gestational hypertension (GHT), gestational proteinuric hypertension (PET), intrauterine growth restriction (IUGR), gestation at delivery, baby's Apgar scores and admission to the neonatal intensive care unit (NICU) were reviewed. Among the 864 patients undergoing IVF treatment, there were 253 live births in total (25 live births in the PCOS group, 54 in the PCO group and 174 in the control group). The prevalence of obstetric complications (GDM, GHT, PET and IUGR) and the obstetric outcomes (gestation at delivery, birth weight, Apgar scores and NICU admissions) were comparable among the three groups. Adjustments for age and multiple pregnancies were made using multiple logistic regression and we found no statistically significant difference among the three groups. Patients with PCO ± PCOS do not have more adverse obstetric outcomes when compared with non-PCO patients undergoing IVF treatment.

  18. Predictors of live birth and pregnancy success after in vitro fertilization in infertile women aged 40 and over.

    Science.gov (United States)

    Kim, Hye Ok; Sung, Nayoung; Song, In Ok

    2017-06-01

    The aim of this study was to evaluate pregnancy outcomes and the live birth rate at 1-year age increments in women aged ≥40 years undergoing fresh non-donor in vitro fertilization (IVF) and embryo transfer (ET), and to identify predictors of success in these patients. This retrospective study was performed among women ≥40 years of age between 2004 and 2011. Of the 2,362 cycles that were conducted, ET was performed in 1,532 (73.1%). The clinical pregnancy rate and live birth rate in women ≥40 years significantly decreased with each year of increased age (plive birth. A statistically significant increase in live birth rates was seen when ≥3 embryos were transferred in patients 40 to 41 years of age, whereas poor pregnancy outcomes were seen in patients ≥43 years of age, regardless of the number of transferred embryos. Moreover, the cumulative live birth rate increased in patients 40 to 42 years of age with repeated IVF cycles, but the follicle-stimulating hormone in those ≥43 years of age rarely showed an increase. IVF-ET has acceptable outcomes in those live birth.

  19. Less depressed or less forthcoming? Self-report of depression symptoms in women preparing for in vitro fertilization

    Science.gov (United States)

    Lewis, Adam M.; Liu, Dawei; Stuart, Scott P.; Ryan, Ginny

    2012-01-01

    PURPOSE While depression has been associated with infertility treatments, it is not routinely assessed in women prior to undergoing in vitro fertilization (IVF) treatment. Findings are mixed regarding the degree to which women report depression prior to IVF. The purpose of this study was to: 1) examine response profiles in women preparing for IVF, and 2) compare responses to those of postpartum, primary care, and general population groups. METHODS Female IVF patients (n=321; 19 – 45 years) completed the PHQ-9 at their first visit. Clinical, demographic characteristics, and incidence of major depressive disorder (MDD) and other depressive disorder (ODD) were examined. Overall score distributions of the IVF group were compared to those of local postpartum patients, and published primary care and general populations. RESULTS Demographic or clinical characteristics did not account for response differences within the IVF group. The IVF group had lower incidences of MDD and ODD than a PHQ-9 normative group. Women in the IVF group reported no depressive symptoms significantly more than postpartum, primary care, and general population groups. CONCLUSIONS Women preparing to undergo IVF report fewer symptoms of depression than multiple comparison groups. Specific quality of life measures may be needed to assess distress in this population. PMID:23138273

  20. The effect of hepatocyte growth factor on mouse oocyte in vitro maturation and subsequent fertilization and embryo development

    Directory of Open Access Journals (Sweden)

    Mohammad H. Bahadori

    2011-05-01

    Full Text Available Background: Oocyte invitro maturation is an enormously promising technology for the treatment of infertility, yet its clinical application remains limited owing to poor success rates. Therefore, this study was devised to evaluate the effect of hepatocyte growth factor (HGF on in vitro maturation of immature mouse oocytes and resulting embryos development. Materials and Method: Cumulus – oocyte complex and germinal vesicle were obtained from eighteen 6-8 weeks-old female NMRI mice 46-48 hours after administration of an injection of 5 IU PMSG (Pregnant Mares’ Serum Gonadotrophin. Oocytes were culture in TCM199 (Tissue culture medium-199 supplemented with dosages of 0, 10, 20, 50 and 100 ng/ml of HGF. After 24 hours, metaphase ІІ oocytes were co-incubated with sperms for 4-6 hours in T6 medium. Following isolation of two pronucleus embryos, cleavage of embryos was assessed in the same medium till blastocyst stage. The number of oocytes and embryos was recorded under an invert microscope and the rate of oocyte maturation, fertilization and embryos cleavage until blastocyst stage compared using of student χ2 test. Results: In all compared groups, oocytes growth and embryos development rate in the 20 ng/ml of HGF treatment group was significantly higher (p<0.05 than the control group (p<0.05.Conclusion: 20 ng/ml of HGF improved the nuclear maturation and embryo development up to blastocyst stage during culture condition

  1. Ectopic pregnancy following in vitro fertilization: meta-analysis and single-center experience during 6 years.

    Science.gov (United States)

    Muller, V; Makhmadalieva, M; Kogan, I; Fedorova, I; Lesik, E; Komarova, E; Dzhemlikhanova, L; Niauri, D; Gzgzyan, A; Ailamazyan, E

    2016-10-01

    Ectopic pregnancy (EP) has been reported to occur in 1.4-5.4% of all clinical pregnancies resulting from in vitro fertilization (IVF) and embryo transfer (ET). Data on factors associated with abnormal embryo implantation following assisted conception are limited. A systematic review and meta-analysis was performed to determine whether there is an association between the day (cleavage-stage, D3, versus blastocyst, D5) or the type (fresh versus frozen/thawed) of ET and EP rate. Risk factors for EP were evaluated in a retrospective study of 1194 women, who achieved pregnancy at our IVF unit between 2010 and 2016. Sixteen papers were considered for the meta-analysis. EP rate did not differ between D3 and D5 fresh ET groups (RR = 0.99, 95%CI: 0.76-1.30) and was higher after fresh versus frozen ET (RR = 1.56, 95%CI: 1.25-1.95). At our clinic, 21 (1.76%) pregnancies were documented as ectopic. The risk of EP was associated with tubal pathology (OR = 3.37, 95%CI: 1.39-8.2), previous appendectomy and past chlamydial infection. Present meta-analysis suggests that EP rate is similar following fresh blastocyst and cleavage ETs, but is significantly reduced after frozen compared with fresh ET. Our own findings demonstrate that tubal pathology has the major impact on EP occurrence following assisted conception.

  2. The design of an in vitro fertilization (IVF laboratory and its importance in risk prevention: applicability of UV radiation

    Directory of Open Access Journals (Sweden)

    Enrique Gea Izquierdo

    2009-12-01

    Full Text Available This study addressed the environmental control of an IVF laboratory and the potential repercussions that the presence of certain physical, chemical and biological agents could have on workers' health or on the tasks they carried out. The design of the laboratory and its facilities contributed to an improvement in work quality and minimised the environmental-related risks. The preventive measures adopted had an influence on the carried out actions.Este estudo dirige-se ao controle ambiental de um laboratório "in vitro fertilization (IVF" e das repercussões potenciais que a presença de determinados agentes físicos, químicos e biológicos possa ter na saúde nos trabalhadores' ou nas tarefas que se realizem. O projeto do laboratório e de suas facilidades contribui a uma melhoria na qualidade do trabalho e minimiza riscos ambiental-relacionados. As medidas preventivas adotadas têm uma influência nas ações realizadas.

  3. Impact of Body Mass Index on Outcomes of In Vitro Fertilization/Intracytoplasmic Sperm Injection Among Polycystic Ovarian Syndrome Patients

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    Na Cui

    2016-10-01

    Full Text Available Background: The aim of this study is to assess the effect of body mass index (BMI on outcomes of in vitro fertilization (IVF / intracytoplasmic sperm injection (ICSI among polycystic ovarian syndrome (PCOS and non-PCOS patients. Methods: This was a retrospective cohort study that was performed in the Second Hospital of Hebei Medical University. Patients who were under 35 years old were included in the study and were divided into four groups based on their BMI. The number of retrieved oocytes, implantation rate, clinical pregnancy rate, miscarriage rate and live births among PCOS and non-PCOS patients were compared between different BMIs. Results: IVF/ICSI pregnancies in obese PCOS women had a considerably higher risk of miscarriage and low rate of clinical pregnancy than in non-obese PCOS pregnancies. However, in non-PCOS patient, obesity significantly elevated miscarriage rate but did not affect clinical pregnancy rate. Conclusion: Obesity in PCOS patients led to poor outcomes of IVF/ICSI.

  4. Predictive value of early serum beta-human chorionic gonadotrophin for the successful outcome in women undergoing in vitro fertilization

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    Neeta Singh

    2013-01-01

    Full Text Available Aims: Pregnancies achieved by in vitro fertilization (IVF are at increased risk of adverse outcome. The main objective of this study was to evaluate the predictive value of β-human chorionic gonadotrophin (β-HCG and age of the patient for the successful outcome in IVF. Materials and Methods: A retrospective study was done in 139 pregnancies after IVF at single IVF center from June 2007 to July 2012. The age of the patient and initial serum values of β-HCG on day 14 of embryo transfer were correlated with ongoing pregnancy (>12 weeks gestation. Results: The β-HCG level on day 14 of more than 347 mIU/ml has a sensitivity of 72.2% and specificity of 73.6% in prediction of pregnancy beyond 12 weeks period of gestation. Positive likelihood ratio (LR is 2.74 and negative LR is 0.37, (receiver operating characteristic area = 0.79. Discussion: In IVF cycles, there is a lot of stress on the couples while the cycle is going on. There was a positive correlation between the higher values of early serum β-HCG levels and ongoing pregnancy. Hence, it can be used as an independent predictor of a successful outcome of IVF cycle. Conclusion: We concluded from our study that early serum β-HCG can be used as a predictor of a successful outcome in IVF.

  5. Predictive value of early serum beta-human chorionic gonadotrophin for the successful outcome in women undergoing in vitro fertilization.

    Science.gov (United States)

    Singh, Neeta; Goyal, Manu; Malhotra, Neena; Tiwari, Abanish; Badiger, Shreenivas

    2013-10-01

    Pregnancies achieved by in vitro fertilization (IVF) are at increased risk of adverse outcome. The main objective of this study was to evaluate the predictive value of β-human chorionic gonadotrophin (β-HCG) and age of the patient for the successful outcome in IVF. A retrospective study was done in 139 pregnancies after IVF at single IVF center from June 2007 to July 2012. The age of the patient and initial serum values of β-HCG on day 14 of embryo transfer were correlated with ongoing pregnancy (>12 weeks gestation). The β-HCG level on day 14 of more than 347 mIU/ml has a sensitivity of 72.2% and specificity of 73.6% in prediction of pregnancy beyond 12 weeks period of gestation. Positive likelihood ratio (LR) is 2.74 and negative LR is 0.37, (receiver operating characteristic area = 0.79). In IVF cycles, there is a lot of stress on the couples while the cycle is going on. There was a positive correlation between the higher values of early serum β-HCG levels and ongoing pregnancy. Hence, it can be used as an independent predictor of a successful outcome of IVF cycle. We concluded from our study that early serum β-HCG can be used as a predictor of a successful outcome in IVF.

  6. Premature luteinization during gonadotropin-releasing hormone antagonist cycles and its relationship with in vitro fertilization outcome.

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    Bosch, Ernesto; Valencia, Iván; Escudero, Ernesto; Crespo, Juana; Simón, Carlos; Remohí, José; Pellicer, Antonio

    2003-12-01

    To determine the prevalence and the effect of premature luteinization in GnRH antagonist IVF-ET cycles. Prospective observational study. In vitro fertilization-embryo transfer (IVF-ET) program at the Instituto Valenciano de Infertilidad. Eighty-one infertile patients undergoing controlled ovarian hyperstimulation with gonadotropins and GnRH antagonist for IVF-ET. Gonadotropin-releasing hormone (GnRH) antagonist was administered from stimulation day 6. Serum P, E(2), and LH were determined on the day of hCG administration. Cycles were grouped according to serum P level on the day of hCG administration ( or =1.2 ng/mL). Clinical pregnancy and implantation rates were determined. The incidence of premature luteinization was 38.3%. Total recombinant FSH dose and stimulation days differed significantly between the groups. Pregnancy rate (25.8% vs. 54.0%) and implantation rate (13.8% vs. 32.0%) were significantly lower in the premature luteinization group. Premature luteinization during GnRH antagonist IVF-ET cycles is a frequent event that is associated with lower pregnancy and implantation rates. Progesterone elevations are not related to serum LH levels and may reflect the mature granulosa cell response to high FSH exposure.

  7. In-vitro fertilization, gamete donation and surrogacy: perceptions of women attending an infertility clinic in Ibadan, Nigeria.

    Science.gov (United States)

    Bello, Folasade A; Akinajo, Opeyemi R; Olayemi, Oladapo

    2014-06-01

    Infertility affects 20% of couples in Nigeria. Assisted reproductive techniques (ART) offered in Nigeria include in-vitro fertilization (IVF), gamete donation and surrogacy. This cross-sectional questionnaire study aimed at assessing the acceptability of ART to women seeking infertility treatment at the University College Hospital, Ibadan, Nigeria. Of the 307 respondents, 58.3% were aware of IVF and 59.3% would accept it as treatment; 35.2% would accept donor eggs and 24.7% would accept donor sperms-a smaller proportion anticipated acceptability by their husbands. Thirty five percent were aware of surrogacy, 37.8% would accept it as treatment; most preferring a stranger as a surrogate. Most felt surrogates should not be paid. Acceptance of ART was associated with older age, longer duration of infertility, previous failed treatment and women without other children. As chances of successful pregnancy are improved in younger individuals, counselling towards overcome barriers to accepting gamete donation and surrogacy should be instituted early.

  8. Current trends of reproductive immunology practices in in vitro fertilization (IVF) - a first world survey using IVF-Worldwide.com.

    Science.gov (United States)

    Kwak-Kim, Joanne; Han, Ae Ra; Gilman-Sachs, Alice; Fishel, Simon; Leong, Milton; Shoham, Zeev

    2013-01-01

    Reproductive immunology has evolved from basic research studies to clinical applications. In this study, we aim to investigate the actual application of reproductive immunology concepts and findings in clinical reproductive medicine such as recurrent pregnancy losses (RPL), repeated implantation failures (RIF), and failed in vitro fertilization (IVF) cycles. A web-based survey was performed on IVF-Worldwide.com. Collected data were analyzed by the computerized software. A significant proportion of physicians recommend thrombophilia workups (86%), parental genetic study (79%), and immunologic evaluations (69%) to IVF candidates who have a history of RPL or chemical pregnancy losses. IVF physicians consider an immunologic workup when patients have two (30%) or three (21%) failed IVF cycles. Assays for anticardiolipin antibody, lupus anticoagulant, thyroid peroxidase antibody, and antinuclear antibody are the four most commonly ordered immunologic tests for RPL (88, 84, 50, 47% each) and RIF (68, 63, 38, 38% each). Cellular immune evaluations, such as NK assay, human leukocyte antigen study, Th1/Th2 study or immunophenotype assay, are less commonly ordered. Reproductive immunology principles have been applied to the clinical management of RPL, RIF, and failed IVF cycles, and a significant proportion of IVF physicians acknowledge the importance of immunologic alterations with reproductive outcomes. © 2012 John Wiley & Sons A/S.

  9. Effect of Fibroblast Co-culture on In Vitro Maturation and Fertilization of Mouse Preantral Follicles

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    Mahmoud Heidari

    2011-01-01

    Full Text Available Background: The aim of this study was to evaluate fibroblast co-culture on in vitro maturation andfertilization of prepubertal mouse preantral follicles.Materials and Methods: The ovaries of 12-14 day old mice were dissected and 120-150 μmintact preantral follicles with one or two layers of granulosa cells, and round oocytes were culturedindividually in α-minimal essential medium (α-MEM supplemented with 5% fetal bovine serum(FBS, 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferrin, seleniummix, 100 μg/ml penicillin and 50 μg/ml streptomycin as base medium for 12 days. A total number of226 follicules were cultured under two conditions: i base medium as control group (n=113; ii basemedium co-cultured with mouse embryonic fibroblast (MEF (n=113. Follicular diameters, alone,in addition to other factors were analyzed by student’s t-test and chi-square test, respectively.Results: The co-culture group showed significant differences (p<0.05 in growth rate (days 4, 6 and8 of the culture period and survival rate. However, there was no significant difference in antrumformation, ovulation rate and embryonic development of released oocytes. There were significantdifferences (p<0.05 in the estradiol and progesterone secretion at all days between the co-cultureand control groups.Conclusion: Fibroblast co-culture increased survival rate and steroid production of preantralfollicles by promoting granulosa cell proliferation.

  10. Demographic determinants and outcome of in vitro fertilization (IVF) services in Saudi Arabia.

    Science.gov (United States)

    Almaslami, Faisal; Aljunid, Syed Mohamed; Ghailan, Khalid

    2018-01-01

    Objectives To assess the demographic characteristics and outcomes of couples undergoing in vitro fertilisation (IVF) treatment at a private hospital in Al Qassim district, Saudi Arabia. Methods For this retrospective study, information was extracted from the hospital electronic database and IVF unit medical records. Results 2259 couples underwent 2703 IVF/ICSI cycles during 2014 to 2016. The utilization rate was approximately 1000 cycle per million of inhabitants. Mean ages ± standard deviation (SD) for women and men were 32.9 ± 5.7 and 39.2 ± 7.4 years, respectively. More couples were diagnosed with secondary infertility (55.2%) than primary infertility (44.8%). Male factor was the commonest single indication for IVF (36.2%). Mean ± SD infertility duration was 4.70 ± 4.03 years. Overall, 949 couples had a successful pregnancy. Age-specific pregnancy rates (PR) were highest for women 40 years (11.6%). Male age and infertility duration had no effect on PR but sperm source (fresh vs. frozen) and female age had significant impacts. However, fresh sperm was used in 90.6% cycles whereas frozen sperm was used in 9.4% cycles. Conclusions IVF treatment outcomes in the Al Qassim district are within the boundaries of average international success rates. Infertile couples seeking IVF services should be counselled with regard to important prognostic factors.

  11. Relationship between Endometrial Thickness and In Vitro Fertilization-Intracytoplasmic Sperm Injection Outcome

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    Turgut Aydin

    2013-01-01

    Full Text Available Background: This study assessed the relationship between endometrial thickness onday of human chorionic gonadotropin (hCG administration and in vitro fertilizationintracytoplasmicsperm injection (IVF-ICSI.Materials and Methods: This prospective cross-sectional study included a total of 593women. Patients were treated with either the agonist or antagonist protocol according to theclinician’s and patient’s preference. Endometrial thickness on the hCG day was measuredby transvaginal ultrasonography (TV-USG. Patients were divided into four groups accordingto endometrial lines, as follows: 14 mm (group 4.Results: Implantation rate (IR, clinical pregnancy rate (CPR, and ongoing pregnancyrate (OPR were significantly lower in group 1 than the other three groups (p<0.05.However, there was no significant difference among groups 2, 3 and 4. Although theendometrial line in the agonist protocol was higher than in the antagonist protocol, thedifference was not statistically significant.Conclusion: The chance of pregnancy appears to be lower in individuals with endometrialthickness less than 7 mm compared with those of higher value.

  12. An increase in IL-1β concentrations in embryo culture-conditioned media obtained by in vitro fertilization on day 3 is related to successful implantation.

    Science.gov (United States)

    Sequeira, Karina; Espejel-Núñez, Aurora; Vega-Hernández, Eva; Molina-Hernández, Anayansi; Grether-González, Patricia

    2015-11-01

    This study aimed to evaluate interleukin (IL)-1β concentrations in maternal serum and in embryo-cultured conditioned media and to correlate these findings with success of implantation. A total of 70 infertile women who underwent in vitro fertilization treatment were studied. IL-1β concentrations were quantified in maternal serum and in embryo culture-conditioned media on days 1 and 3. The findings were compared between those who achieved pregnancy and those who did not. No significant differences were found in IL-1β serum concentrations between the groups. IL-1β was not detected in day 1 culture-conditioned medium. On day 3, IL-1β was quantified in 27 patients, and IL-1β concentrations were significantly higher in women who achieved pregnancy than in those who did not (p culture-conditioned medium in patients who achieve pregnancy after in vitro fertilization treatment indicate a possible role of embryonic IL-1β in the implantation process.

  13. The influences of GnRH, oxytocin and vasoactive intestinal peptide on LH and PRL secretion by porcine pituitary cells in vitro.

    Science.gov (United States)

    Bogacka, I; Siawrys, G; Okrasa, S; Kaminski, T; Przala, J

    2002-09-01

    (Groups 2 and 3) or P4. In contrast, in OVX gilts primed with placebo, VIP was without any effect on PRL secretion. In conclusion, the results of our in vitro studies confirmed the stimulatory effect of GnRH on LH secretion by porcine pituitary cells and also suggest a participation of OT and VIP in modulation of LH and PRL secretion at the pituitary level in a way dependent on hormonal status of animals.

  14. Absence of hepatitis B virus (HBV) DNA in children born after exposure of their mothers to HBV during in vitro fertilization.

    Science.gov (United States)

    Quint, W G; Fetter, W P; van Os, H C; Heijtink, R A

    1994-01-01

    During in vitro fertilization, 22 human embryos were exposed to hepatitis B virus (HBV) in contaminated human serum present in the culture medium. All mothers experienced hepatitis B during the first trimester of pregnancy, and two had hepatitis B surface antigen and HBV DNA, as determined by PCR, at the time of delivery. No HBV DNA was found in serum or lymphocytes from the exposed 22 infants. HBV DNA was also absent from one infant at autopsy. PMID:8027321

  15. In vitro fertilization and artificial activation of eggs of the direct-developing anuran Eleutherodactylus coqui

    OpenAIRE

    Toro Esteban; Michael Scott F

    2004-01-01

    Abstract Although much is known about the reproductive biology of pond-breeding frogs, there is comparatively little information about terrestrial-breeding anurans, a highly successful and diverse group. This study investigates the activation and in vitro fertilization of eggs of the Puerto Rican coqui frog obtained by hormonally induced ovulation. We report that spontaneous activation occurs in 34% of eggs, probably in response to mechanical stress during oviposition. Artificial activation, ...

  16. Hormonal priming, induction of ovulation and in-vitro fertilization of the endangered Wyoming toad (Bufo baxteri)

    Science.gov (United States)

    Browne, Robert K; Seratt, Jessica; Vance, Carrie; Kouba, Andrew

    2006-01-01

    The endangered Wyoming toad (Bufo baxteri) is the subject of an extensive captive breeding and reintroduction program. Wyoming toads in captivity rarely ovulate spontaneously and hormonal induction is used to ovulate females or to stimulate spermiation in males. With hormonal induction, ovulation is unreliable and egg numbers are low. The sequential administration of anovulatory doses of hormones (priming) has increased egg numbers and quality in both anurans and fish. Consequently, we tested the efficacy of a combination of human Chorionic Gonadotrophin (hCG) and Luteinizing Hormone Releasing Hormone analogue (LHRHa) administered as one dose, or two or three sequential doses to Bufo baxteri on egg numbers, fertilization and early embryo development. Spawning toads deposited eggs into Simplified Amphibian Ringers (SAR) solution to enable controlled in-vitro fertilization (IVF) with sperm from hormonally induced male toads. Unprimed females receiving a single mixed normally ovulatory dose of 500 IU hCG plus 4 micrograms of LHRHa produced no eggs. Whereas females primed with this dose and an anovulatory dose (100 IU hCG and 0.8 micrograms of LHRHa) of the same hormones, or primed only with an anovulatory dose, spawned after then receiving an ovulatory dose. Higher total egg numbers were produced with two primings than with one priming. Moreover, two primings produced significantly more eggs from each individual female than one priming. The cleavage rate of eggs was not found to differ between one or two primings. Nevertheless, embryo development with eggs from two primings gave a significantly greater percentage neurulation and swim-up than those from one priming. Of the male toads receiving a single dose of 300 IU hCG, 80% produced spermic urine with the greatest sperm concentration 7 hours post-administration (PA). However, peak sperm motility (95%) was achieved at 5 hours PA and remained relatively constant until declining 20 hours PA. In conclusion, Bufo baxteri

  17. Clinical review of 24-35 year olds conceived with and without in vitro fertilization: study protocol.

    Science.gov (United States)

    Lewis, Sharon; Kennedy, Joanne; Burgner, David; McLachlan, Robert; Ranganathan, Sarath; Hammarberg, Karin; Saffery, Richard; Amor, David J; Cheung, Michael M H; Doyle, Lex W; Juonala, Markus; Donath, Susan; McBain, John; Halliday, Jane

    2017-09-20

    Children conceived by assisted reproductive technologies (ART) currently comprise 4% of Australian births. The manipulation of biological parameters related to fertilization and implantation are integral to successful ART but potentially pose a risk to the longer-term health of the offspring. There is consensus that many common adult health problems (particularly cardiovascular, metabolic and respiratory conditions) have their origins in early life, possibly before birth, and that risk trajectories track through childhood until clinical disease manifests in adulthood. Early life epigenetic variation may play a role in this process. However little is known about the long-term health of individuals conceived by ART. In a previous study, based on telephone-interviews, we found that young adults conceived by in vitro fertilization (IVF) had significantly more maternal reported atopic respiratory, endocrine, nutritional, and metabolic conditions than non-IVF conceived matched controls. Here we outline the protocol for a follow-up biomedical assessment of this cohort and a questionnaire to obtain information on potential confounders. We are conducting a clinical review of an existing, well characterised cohort comprising 547 IVF-conceived adults and 549 matched controls. We are measuring cardiovascular intermediate phenotypes, metabolic parameters and respiratory function, complemented by epigenome-wide DNA methylation analysis. A pilot study demonstrated the feasibility of our proposed protocol and its acceptability to participants. Participants attend a 2-3 h clinical assessment and complete a study-specific online questionnaire. Measurements include: 1) cardiovascular phenotypes: carotid artery intima-media thickness and distensibility, retinal vascular calibre, resting blood pressure, pulse wave velocity and pulse wave analysis; 2) respiratory function: spirometry, plethysmography, multiple breath washout; 3) auxology: height, weight, waist circumference, bio

  18. Effect of rapamycin treatment during post-activation and/or in vitro culture on embryonic development after parthenogenesis and in vitro fertilization in pigs.

    Science.gov (United States)

    Elahi, F; Lee, H; Lee, J; Lee, S T; Park, C K; Hyun, S-H; Lee, E

    2017-10-01

    This study investigated the effects of early induction of autophagy on embryonic development in pigs. For this, oocytes or embryos were treated with an autophagy inducer, rapamycin (RP), during post-activation (Pa), in vitro fertilization (IVF) and/or in vitro culture (IVC). When parthenogenesis (PA) embryos were untreated (control) or treated with various concentrations of RP for 4 hr during Pa, 100 nm RP showed a higher blastocyst formation (48.8 ± 2.7%) than the control (34.6 ± 3.0%). When PA embryos were treated during the first 24 hr of IVC, blastocyst formation was increased (p < .05) by 1 and 10 nm RP (61.9 ± 3.0 and 59.6 ± 3.0%, respectively) compared to the control (43.2 ± 1.8%) and 100 nm RP (47.8 ± 3.2%), with a higher embryo cleavage in response to 10 nm RP (87.3 ± 2.4%) than the control (74.1 ± 3.2%). RP treatment during IVC and Pa + IVC showed increased blastocyst formation (44.7 ± 2.5 and 44.1 ± 2.0%, respectively) compared to the control (33.2 ± 2.0%). In addition, RP treatment during Pa and/or IVC increased glutathione content and inversely reduced reactive oxygen species. In IVF, RP treatment for 6 hr during IVF significantly increased embryonic development (34.0 ± 2.6%) compared to the control (24.8 ± 1.6%), but treatment during IVC for 24 hr with RP did not (23.0 ± 3.8%). Autophagy was significantly increased in PA oocytes by the RP treatment during Pa but not altered by the treatment during the first 24 hr of IVC. Overall, RP treatment positively regulated the pre-implantation development of pig embryos, probably by regulating cellular redox state and stimulating autophagy. © 2017 Blackwell Verlag GmbH.

  19. Transcription Adaptation during In Vitro Adipogenesis and Osteogenesis of Porcine Mesenchymal Stem Cells: Dynamics of Pathways, Biological Processes, Up-Stream Regulators, and Gene Networks.

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    Massimo Bionaz

    Full Text Available The importance of mesenchymal stem cells (MSC for bone regeneration is growing. Among MSC the bone marrow-derived stem cells (BMSC are considered the gold standard in tissue engineering and regenerative medicine; however, the adipose-derived stem cells (ASC have very similar properties and some advantages to be considered a good alternative to BMSC. The molecular mechanisms driving adipogenesis are relatively well-known but mechanisms driving osteogenesis are poorly known, particularly in pig. In the present study we have used transcriptome analysis to unravel pathways and biological functions driving in vitro adipogenesis and osteogenesis in BMSC and ASC. The analysis was performed using the novel Dynamic Impact Approach and functional enrichment analysis. In addition, a k-mean cluster analysis in association with enrichment analysis, networks reconstruction, and transcription factors overlapping analysis were performed in order to uncover the coordination of biological functions underlining differentiations. Analysis indicated a larger and more coordinated transcriptomic adaptation during adipogenesis compared to osteogenesis, with a larger induction of metabolism, particularly lipid synthesis (mostly triglycerides, and a larger use of amino acids for synthesis of feed-forward adipogenic compounds, larger cell signaling, lower cell-to-cell interactions, particularly for the cytoskeleton organization and cell junctions, and lower cell proliferation. The coordination of adipogenesis was mostly driven by Peroxisome Proliferator-activated Receptors together with other known adipogenic transcription factors. Only a few pathways and functions were more induced during osteogenesis compared to adipogenesis and some were more inhibited during osteogenesis, such as cholesterol and protein synthesis. Up-stream transcription factor analysis indicated activation of several lipid-related transcription regulators (e.g., PPARs and CEBPα during adipogenesis

  20. Cyclic test comparison of all-inside device and inside-out sutures for radial meniscus lesion repair: an in vitro porcine model study.

    Science.gov (United States)

    Lee, Yee Han Dave; Nyland, John; Burden, Robert; Caborn, David N M

    2012-12-01

    To compare biomechanical fixation and gapping characteristics of a new all-inside meniscus repair method for radial meniscus lesion repair versus conventional inside-out suture repair under submaximal cyclic loading and load-to-failure test conditions. Fresh-frozen porcine tibiae with attached lateral menisci and joint capsules were harvested and stored for 48 hours at -20°C. After thawing for 12 hours, equivalent-size healthy specimens were randomly assigned to 2 groups of 8 specimens each. Standardized radial lesions were repaired with the Sequent device (ConMed Linvatec, Largo, FL) (group 1) or conventional inside-out suturing with No. 2-0 braided polyester suture (group 2). Repaired specimens were placed in custom clamps and mounted on a servohydraulic device. After a 2-N preload, specimens were cycled from 5 to 20 N (0.1 Hz), before undergoing 1,000 submaximal loading cycles between 5 and 20 N (0.5 Hz). A 40-second delay at 100, 500, and 1,000 cycles enabled digital photographs to be taken for gapping measurement determination. Specimens then underwent load-to-failure testing (12.5 mm/s). Fixation failure mode was documented. Group displacement did not differ after 1, 100, 500, and 1,000 submaximal loading cycles. Group peak gapping did not differ at 100, 500, and 1,000 submaximal loading cycles. Load at failure and displacement and stiffness during load-to-failure testing did not differ between groups. During load-to-failure testing, all-inside specimens failed by implant dislodgement from the meniscus periphery whereas the inside-out repaired specimens failed by suture rupture. Under controlled in vitro biomechanical test conditions, the all-inside device provided radial meniscus lesion fixation that was comparable, but not superior, to conventional inside-out suturing. The all-inside radial lateral meniscus lesion repair method may provide comparable fixation to conventional inside-out suturing without the need for additional incisions and their associated

  1. Investigating the Role of Surface Materials and Three Dimensional Architecture on In Vitro Differentiation of Porcine Monocyte-Derived Dendritic Cells.

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    Sofie Bruun Hartmann

    Full Text Available In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured in materials other than polystyrene and applying three-dimensional structures more similar to the in vivo environment. Polydimethylsiloxane (PDMS is an often used polymer for lab-on-a-chip devices but not much is known about the effect of changing the culture surface material from polystyrene to PDMS. In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material and architecture. Distinct clustering of cells, based on similar expression patterns of 46 genes of interest, was seen for cells isolated from two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS. Changing the material from polystyrene to PDMS resulted in cells with expression patterns usually associated with macrophage expression (upregulation of CD163 and downregulation of CD1a, FLT3, LAMP3 and BATF3. However, this was purely based on gene expression level, and no functional assays were included in this study which would be necessary in order to classify the cells as being macrophages. When changing to three-dimensional culture the cells became increasingly activated in terms of IL6, IL8, IL10 and CCR5 gene expression. Further stimulation

  2. 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.

  3. 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

  4. Association of in vitro fertilization with global and IGF2/H19 methylation variation in newborn twins.

    Science.gov (United States)

    Loke, Y J; Galati, J C; Saffery, R; Craig, J M

    2015-04-01

    In vitro fertilization (IVF) and its subset intracytoplasmic sperm injection (ICSI), are widely used medical treatments for conception. There has been controversy over whether IVF is associated with adverse short- and long-term health outcomes of offspring. As with other prenatal factors, epigenetic change is thought to be a molecular mediator of any in utero programming effects. Most studies focused on DNA methylation at gene-specific and genomic level, with only a few on associations between DNA methylation and IVF. Using buccal epithelium from 208 twin pairs from the Peri/Postnatal Epigenetic Twin Study (PETS), we investigated associations between IVF and DNA methylation on a global level, using the proxies of Alu and LINE-1 interspersed repeats in addition to two locus-specific regulatory regions within IGF2/H19, controlling for 13 potentially confounding factors. Using multiple correction testing, we found strong evidence that IVF-conceived twins have lower DNA methylation in Alu, and weak evidence of lower methylation in one of the two IGF2/H19 regulatory regions and LINE-1, compared with naturally conceived twins. Weak evidence of a relationship between ICSI and DNA methylation within IGF2/H19 regulatory region was found, suggesting that one or more of the processes associated with IVF/ICSI may contribute to these methylation differences. Lower within- and between-pair DNA methylation variation was also found in IVF-conceived twins for LINE-1, Alu and one IGF2/H19 regulatory region. Although larger sample sizes are needed, our results provide additional insight to the possible influence of IVF and ICSI on DNA methylation. To our knowledge, this is the largest study to date investigating the association of IVF and DNA methylation.

  5. Opportunities for reproductive tourism: cost and quality advantages of Turkey in the provision of in-vitro Fertilization (IVF) services.

    Science.gov (United States)

    Yildiz, M Said; Khan, M Mahmud

    2016-08-12

    The scale and scope of medical tourism have expanded rapidly over the last few decades. Turkey is becoming an important player in this market because of its relatively better service quality and large comparative cost advantage. This paper compares cost, quality and effectiveness of in-vitro fertilization (IVF) in the USA and in Turkey. The data from Turkey were obtained from a hospital specializing in IVF services and the US data came from secondary sources. Package price offered by the dominant IVF-service provider to international patients in Turkey was used as a measure of cost for Turkey while IVF-specific service prices were used to estimate the cost for USA. To compare quality and effectiveness of IVF services, a number of general clinical quality indicators and IVF success rate were used. Indicators of quality, cost and success rate in the Turkish hospital were found to be better than the corresponding indicators in US hospitals. The cost difference of IVF services between USA and Turkey is so significant that the overall cost of obtaining the service from Turkey remains lower even with additional expenses for travel and accommodation. Cost-effectiveness ratio of IVF treatment per successful clinical pregnancy was much lower in Turkey than in the USA. It appears that cost and quality are the two most important factors affecting demand for health care services by international patients in Turkey. Like other important players in the medical tourism market, Turkey should be able to take advantage of its success in IVF, a highly specialized niche market, to transform its health system into an important exporter of general health services.

  6. Results of gonadotrophin stimulation with the option to convert cycles to in vitro fertilization in cases of multifollicular development.

    Science.gov (United States)

    Bergh, C; Bryman, I; Nilsson, L; Janson, P O

    1998-01-01

    To avoid a high cancellation rate and/or a high multiple pregnancy rate due to multifollicular development in gonadotrophin stimulated cycles, such cycles were converted in the same cycle to in vitro fertilization/embryo transfer (IVF/ET). The results from a four year period using this strategy are summarized. Seventy-three anovulatory women (seven WHO group I, 66 WHO group II) were studied during this period. In a majority of the cycles a GnRH-analogue was used for down-regulation according to a long protocol, followed by stimulation with FSH and/or hMG. Out of 154 WHO group II gonadotrophin stimulation cycles intended for ovulation induction, 25 cycles were converted to IVF. The pregnancy and delivery rates in the IVF-converted cycles were 50% and 41%, respectively, and 31% and 22% when gonadotrophin stimulation was followed by intercourse. The cancellation rate, including both ovulation induction and IVF cycles, was 15% and the multiple pregnancy rate was 30%, mainly twins. Lean women achieved better outcome than obese women. In WHO group I only 12 cycles were performed. One cycle was converted to IVF resulting in delivery and one cycle was cancelled. The pregnancy- and delivery rates were both 50% when gonadotrophin stimulation was followed by intercourse. It is concluded that the option to convert a gonadotrophin stimulation cycle to IVF in the same cycle, in cases of multifollicular development, seemed to be a good alternative. The conversion results in a low cancellation rate and a low incidence of high order multiple pregnancies. Patients should be informed of this opportunity before entering ovulation stimulation.

  7. Effect of intramural fibroid on uterine and endometrial vascularity in infertile women scheduled for in-vitro fertilization.

    Science.gov (United States)

    Kamel, Ahmed; El-Mazny, Akmal; Ramadan, Wafaa; Abdelaziz, Suzy; Gad-Allah, Sherine; Saad, Hany; Hussein, Ahmed M; Salah, Emad

    2018-02-01

    To study the effect of intramural fibroids on uterine and endometrial vascularity in infertile women scheduled for in-vitro fertilization (IVF). 3D power Doppler was used to measure the endometrial volume and blood flow indices in 182 women with intramural fibroids not affecting the uterine cavity and compared them to a matched control group without fibroids. There was significantly increased vascularity in the endometrium of the fibroid group as denoted by higher endometrial VI (p = 0.018), FI (p = 0.027) and Endometrial VFI. No significant difference in mean uterine artery RI (p = 0.277) or PI (p = 0.187). Among the fibroid group 62.6% had a fibroid > 4 cm. Women with fibroids > 4 cm had a significantly higher Endometrial FI (p = 0.037), and VFI (p = 0.02). Uterine artery blood flow was not affected, as uterine RI (p = 0.369) and PI (p = 0.321) were not statistically different. Compared with the control group (non fibroid), women with fibroids > 4 cm had significantly higher endometrial VI (p = 0.013), FI (p = 0.004), and VFI (p  4 cm significantly increase endometrial vascularity. This increase in blood flow may be a factor that affects the outcome of IVF.

  8. A logistic model to predict early pregnancy loss following in vitro fertilization based on 2601 infertility patients.

    Science.gov (United States)

    Yi, Yan; Lu, Guangxiu; Ouyang, Yan; lin, Ge; Gong, Fei; Li, Xihong

    2016-03-31

    According to previous studies, even after embryonic cardiac activity is detected, the pregnancy loss rate remains 3-4 %. The objectives of this study were to investigate the differences in ultrasound parameters between a miscarriage group and an ongoing pregnancy group during the 1(st) trimester and to build a logistic model to predict early pregnanc