Action of uranium on pre implanted mouse embryos

International Nuclear Information System (INIS)

Kundt, Miriam S.

2001-01-01

The cultured preimplantation embryos are normally employed to evaluate the effects of environmental pollutants specially metals. Embryos were obtained from hybrid females CBA x C57 Bl following induction of super ovulation. They were incubated from 1 cell stage during 120 hs. in M16 cultured medium. Three different experiments were carried out: A, B and C using uranyl nitrate UO 2 (NO 3 ) 2 6H 2 O as source of uranium. In experiment 'A' the embryos were cultivated in the same culture dish containing final U concentrations of 13, 26, 52, 104 and 208 μgU/ml. In experiment 'B' embryos in a one cell stage were placed in culture medium with uranyl nitrate with final U concentrations of 26, 52, 104 μgU/ml. After 24 hours those embryos which had reached the two-cell stage were transferred to another culture dish to which fresh solutions of uranyl nitrate were added, maintaining the same concentrations of the previous one. In experiment 'C' the embryos were cultivated containing final U concentrations of 26, 52 and 104 μgU/ml and they were transferred to another culture dish every day to which fresh solutions of uranyl nitrate were added. Different embryos parameters were analyzed: 1) Development grade; 2) Number of cell per embryo and metaphases index; and 3) Embryo ploidy. 1) Embryos were observed each 24 hs. to evaluate development grade: 2, 4 and 8 cell stage, morula, early -expanded- hatched blastocysts and atresic embryos. No significant differences were observed in the proportion of embryos arrested either in the one-cell or in the two cell stages in control culture medium regarding different concentrations of U, in a total of 4388 embryos analyzed. From 2 cell stage, moment that the embryo begins to synthesize its own ARNm, the delay in embryonic development increased dose dependent. On the other hand, the toxicological effects in the same concentration are increase from 'A' treatment to 'C' treatment. Embriotoxicology effects are evidenced by an increment in

The endometrial factor in human embryo implantation

NARCIS (Netherlands)

Boomsma, C.M.

2009-01-01

The studies presented in this thesis aimed to explore the role of the endometrium in the implantation process. At present, embryo implantation is the major rate-limiting step for success in fertility treatment. Clinicians have sought to develop clinical interventions aimed at enhancing implantation

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

Science.gov (United States)

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

2015-11-01

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

Sexing bovine pre-implantation embryos using the polymerase ...

African Journals Online (AJOL)

The paper aims to present a bovine model for human embryo sexing. Cows were super-ovulated, artificially inseminated and embryos were recovered 7 days later. Embryo biopsy was performed; DNA was extracted from blastomeres and amplified using bovine-specific and bovine-Y-chromosomespecific primers, followed ...

NMR studies of preimplantation embryo metabolism in human assisted reproductive techniques: a new biomarker for assessment of embryo implantation potential.

Science.gov (United States)

Pudakalakatti, Shivanand M; Uppangala, Shubhashree; D'Souza, Fiona; Kalthur, Guruprasad; Kumar, Pratap; Adiga, Satish Kumar; Atreya, Hanudatta S

2013-01-01

There has been growing interest in understanding energy metabolism in human embryos generated using assisted reproductive techniques (ART) for improving the overall success rate of the method. Using NMR spectroscopy as a noninvasive tool, we studied human embryo metabolism to identify specific biomarkers to assess the quality of embryos for their implantation potential. The study was based on estimation of pyruvate, lactate and alanine levels in the growth medium, ISM1, used in the culture of embryos. An NMR study involving 127 embryos from 48 couples revealed that embryos transferred on Day 3 (after 72 h in vitro culture) with successful implantation (pregnancy) exhibited significantly (p < 10(-5) ) lower pyruvate/alanine ratios compared to those that failed to implant. Lactate levels in media were similar for all embryos. This implies that in addition to lactate production, successfully implanted embryos use pyruvate to produce alanine and other cellular functions. While pyruvate and alanine individually have been used as biomarkers, the present study highlights the potential of combining them to provide a single parameter that correlates strongly with implantation potential. Copyright © 2012 John Wiley & Sons, Ltd.

The effect of valproic acid and oxcarbazepine on the distribution of adhesion molecules in embryo implantation

International Nuclear Information System (INIS)

Gürgen, Seren Gülşen; Erdoğan, Deniz; Coşkun, Zafer Kutay; Cansu, Ali

2012-01-01

This study was intended to investigate the effect of valproate (VPA) and oxcarbazepine (OXC) on embryo implantation in terms of extracellular matrix protein distribution. Thirty female rats (Wistar albino) were assigned to three groups of 10 animals each. Group 1 was administered two doses of saline solution, group 2, two doses of VPA at 300 mg/kg/day and group 3, two doses of OXC at 100 mg/kg/day, for a period of 3 months. Female rats with vaginal plugs mated with males for one night were placed into separate cages. Day of mating was taken as day 0, and implantation areas were obtained with rats being sacrificed on the morning of day 7. Immunohistochemical staining and electron microscopic protocols were then applied. At electron microscopic evaluation, extraembryonic endoderm and ectoderm layers could not be distinguished in semi-thin sections in the VPA group, while they were partially differentiated in the OXC group. At immunohistochemical staining, laminin was observed in the primary embryonic endoderm cell visceral and parietal layers, the uterine luminal epithelial cells and the secondary decidual zone in the control group. In the VPA group, it was weakly expressed in some embryo trophoectoderm cells and uterine luminal epithelial cells and moderately in some decidual cells. In the OXC group, it was moderately expressed in some trophoectoderm and decidual cells. Collagen IV was localized in the ectoplacental cone cells and secondary decidual zone and weak in the luminal epithelial cells in the control group. In the VPA and OXC groups, collagen IV was negative in all embryonic and maternal structures in the VPA and OXC groups. Vimentin was moderately expressed in the luminal epithelium and strongly expressed in the primary decidual zone and ectoplacental cone cells in the control group. In the VPA group, it was negative in the embryo trophoectoderm, decidual and uterine luminal epithelial cells, while in the OXC group it was moderately localized in the

Expression of proposed implantation marker genes CDX2 and HOXB7 in the blastocyst does not distinguish viable from non-viable human embryos

DEFF Research Database (Denmark)

Kirkegaard, Kirstine; Hindkjær, Johnny Juhl; Ingerslev, Hans Jakob

2012-01-01

expression differs between viable and non-viable embryos in both human and non-humans, suggesting transcriptome analysis of trophectoderm (TE) as a novel method of improving embryo selection. Potential candidate marker genes have been identified with array studies on animal blastocysts. The aim of this study...... was to investigate the expression of selected genes in human blastocysts in relation to the outcome of implantation. Materials and methods: Embryos from 10 oatients undergoing in vitro fertilization treatment were included in the project. A single blastocyst was chosen for biopsy on the morning of day 5 after oocyte...... of 15 key genes associated with developmental competence in animals were evaluated in high quality human embryos with monogenic or chromosomal disorders from a pre-implantation genetic disorder program. Triplicate cDNA amplifications for quantitative (q) RT-PCR were performed using pre-designed gene...

Efficacy of postal communication with patients who have cryopreserved pre-embryos.

Science.gov (United States)

Brzyski, R G

1998-11-01

To compare the characteristics of patients who did and did not respond to a request for information regarding their cryopreserved pre-embryos. Mail survey. Academic-assisted reproductive technology program. One hundred thirty-six patients with cryopreserved pre-embryos. Patients were surveyed by first-class mail regarding their plans for their cryopreserved pre-embryos and their interest in embryo donation. Age, number of stored pre-embryos, and duration of storage of responders and nonresponders at 6 weeks after mailing. Eighty-three patients (62%) did not respond to the survey. Compared with responders, nonresponders were significantly older at the time of embryo cryopreservation, had fewer pre-embryos cryopreserved, and had the pre-embryos cryopreserved for a longer duration. Five responders (9%) expressed an interest in embryo donation. Three patients requested disposal of pre-embryos. Sixteen surveys (12%) were returned as undeliverable. As a group, these patients had the fewest pre-embryos cryopreserved and had the longest duration of storage. A disturbing number of patients with cryopreserved pre-embryos ignored efforts by our program to maintain contact. Older patients with few cryopreserved pre-embryos may require special attention to avoid abandonment.

Embryo implantation: Shedding light on the roles of ovarian ...

African Journals Online (AJOL)

Implantation is a crucial step in mammalian reproduction, as it is a gateway to further embryonic development and successful pregnancy. Successful implantation requires coordinated interactions between the blastocyst and uterus. Uterine receptivity for embryo implantation is regulated by the ovarian hormones estrogen ...

Insights from imaging the implanting embryo and the uterine environment in three dimensions

Science.gov (United States)

Arora, Ripla; Fries, Adam; Oelerich, Karina; Marchuk, Kyle; Sabeur, Khalida; Giudice, Linda C.

2016-01-01

Although much is known about the embryo during implantation, the architecture of the uterine environment in which the early embryo develops is not well understood. We employed confocal imaging in combination with 3D analysis to identify and quantify dynamic changes to the luminal structure of murine uterus in preparation for implantation. When applied to mouse mutants with known implantation defects, this method detected striking peri-implantation abnormalities in uterine morphology that cannot be visualized by histology. We revealed 3D organization of uterine glands and found that they undergo a stereotypical reorientation concurrent with implantation. Furthermore, we extended this technique to generate a 3D rendering of the cycling human endometrium. Analyzing the uterine and embryo structure in 3D for different genetic mutants and pathological conditions will help uncover novel molecular pathways and global structural changes that contribute to successful implantation of an embryo. PMID:27836961

[Conception rate and embryo development in guinea pigs with synchronized estrus induced by progesterone implant].

Science.gov (United States)

Ueda, H; Kosaka, T; Takahashi, K W

1994-01-01

Observations were made on the timing of mating and the pre-implantation development of fertilized eggs in guinea pigs synchronized by long-term progesterone treatment. Females received a subcutaneous implant of progesterone-filled silastic tubing for 14 days. Copulation was observed from the evening of day 4 to the morning of day 6 in 53 of 54 females (98%). Most of them (47/53, 89%) copulated on day 5 after removal of the tubing. Designating the day of copulation (day 5 after removal of the tubing) as day 0 of gestation, embryos collected from the genital tract were at the 4-cell, 8-cell, morula, and blastocyst stages on days 1, 3, 4 and 5 of gestation, respectively. Eggs were recovered at high incidence (85-100%) from days 1 to 5 of gestation. On day 6 gestation, no eggs were recovered from the genital tract, suggesting that implantation had occurred. The mean litter size (+/- S. D.) was 4.0 +/- 0.8 pups, which were born normally after a mean gestation period of 67 +/- 1 days in 7 synchronized females. Since the female guinea pigs synchronized by the long-term progesterone treatment had normal reproductive ability similar to that of cyclic females, this technique would make it possible to obtain animals at a scheduled time even in smaller-sized colonies. In addition, observations on the pre-implantation development of embryos in females with synchronized estrus might be a useful aid in the field of reproductive research.

Endometrial signals improve embryo outcome: functional role of vascular endothelial growth factor isoforms on embryo development and implantation in mice.

Science.gov (United States)

Binder, N K; Evans, J; Gardner, D K; Salamonsen, L A; Hannan, N J

2014-10-10

Does vascular endothelial growth factor (VEGF) have important roles during early embryo development and implantation? VEGF plays key roles during mouse preimplantation embryo development, with beneficial effects on time to cavitation, blastocyst cell number and outgrowth, as well as implantation rate and fetal limb development. Embryo implantation requires synchronized dialog between maternal cells and those of the conceptus. Following ovulation, secretions from endometrial glands increase and accumulate in the uterine lumen. These secretions contain important mediators that support the conceptus during the peri-implantation phase. Previously, we demonstrated a significant reduction of VEGFA in the uterine cavity of women with unexplained infertility. Functional studies demonstrated that VEGF significantly enhanced endometrial epithelial cell adhesive properties and embryo outgrowth. Human endometrial lavages (n = 6) were obtained from women of proven fertility. Four-week old Swiss mice were superovulated and mated with Swiss males to obtain embryos for treatment with VEGF in vitro. Preimplantation embryo development was assessed prior to embryo transfer (n = 19-30/treatment group/output). Recipient F1 female mice (8-12 weeks of age) were mated with vasectomized males to induce pseudopregnancy and embryos were transferred. On Day 14.5 of pregnancy, uterine horns were collected for analysis of implantation rates as well as placental and fetal development (n = 14-19/treatment). Lavage fluid was assessed by western immunoblot analysis to determine the VEGF isoforms present. Mouse embryos were treated with either recombinant human (rh)VEGF, or VEGF isoforms 121 and 165. Preimplantation embryo development was quantified using time-lapse microscopy. Blastocysts were (i) stained for cell number, (ii) transferred to wells coated with fibronectin to examine trophoblast outgrowth or (iii) transferred to pseudo pregnant recipients to analyze implantation rates, placental and

Contribution of immunology to implantation failure of euploid embryos.

Science.gov (United States)

Franasiak, Jason M; Scott, Richard T

2017-06-01

Outcomes in assisted reproduction have seen marked improvement. With increased ability in the embryology laboratory to use extended embryo culture which in turn enables other selective techniques, such as trophectoderm biopsy and comprehensive chromosome screening, the chance of success per embryo transfer is increased. However, even the selection of a euploid blastocyst, which selects out many embryonic factors, does not yield successful implantation and ultimately delivery in all cases. Among the factors that affect implantation failure of apparently reproductively competent embryos, the immune system has been perhaps both the most plausible and the most debated. There are data on T-helper cells, in particular the T H 1-T H 2 balance, peripheral and uterine natural killer cells, and autoantibodies, all of which have been shown to have variable effects on implantation. Many investigators have developed and used a wide range of immune tests and treatments aimed at manipulating the milieu to favor implantation. Although it is certain that the immune system plays a role in implantation, our understanding of the physiology, let alone the pathophysiology, remains incomplete. It is imperative that we gain more clear evidence of causes and test and implement treatment paradigms. In the meantime, immune testing or empirical treatment with the use of immune modulators must be approached with caution. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Inverted light-sheet microscope for imaging mouse pre-implantation development.

Science.gov (United States)

Strnad, Petr; Gunther, Stefan; Reichmann, Judith; Krzic, Uros; Balazs, Balint; de Medeiros, Gustavo; Norlin, Nils; Hiiragi, Takashi; Hufnagel, Lars; Ellenberg, Jan

2016-02-01

Despite its importance for understanding human infertility and congenital diseases, early mammalian development has remained inaccessible to in toto imaging. We developed an inverted light-sheet microscope that enabled us to image mouse embryos from zygote to blastocyst, computationally track all cells and reconstruct a complete lineage tree of mouse pre-implantation development. We used this unique data set to show that the first cell fate specification occurs at the 16-cell stage.

Embryo quality and impact of specific embryo characteristics on ongoing implantation in unselected embryos derived from modified natural cycle in vitro fertilization

NARCIS (Netherlands)

Pelinck, Marie-Jose; Hoek, Annemieke; Simons, Arnold H. M.; Heineman, Maas Jan; van Echten-Arends, Janny; Arts, Eus G. J. M.

Objective: To study the implantation potential of unselected embryos derived from modified natural cycle IVF according to their morphological characteristics. Design: Cohort study. Setting: Academic department of reproductive medicine. Patient(S): A series of 449 single embryo transfers derived from

A multicenter prospective study to assess the effect of early cleavage on embryo quality, implantation, and live-birth rate.

Science.gov (United States)

de los Santos, Maria José; Arroyo, Gemma; Busquet, Ana; Calderón, Gloria; Cuadros, Jorge; Hurtado de Mendoza, Maria Victoria; Moragas, Marta; Herrer, Raquel; Ortiz, Agueda; Pons, Carme; Ten, Jorge; Vilches, Miguel Angel; Figueroa, Maria José

2014-04-01

To investigate the impact of early cleavage (EC) on embryo quality, implantation, and live-birth rates. Prospective cross-sectional study. Multicenter study. Seven hundred embryo transfers and 1,028 early-stage human embryos. None. Implantation according to the presence of EC and embryo quality. The presence of EC is associated with embryo quality, especially in cycles with autologous oocytes. However, the use of EC as an additional criterion for selecting an embryo for transfer does not appear to significantly improve likelihood of implantation. Furthermore, embryos that presented EC had live-birth rates per implanted embryo similar to those that did not show any sign of cleavage. At least for conventional embryo culture and morphologic evaluations, the additional evaluation of EC in embryos may not be valuable to improve embryo implantation. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Embryo-epithelium interactions during implantation at a glance.

Science.gov (United States)

Aplin, John D; Ruane, Peter T

2017-01-01

At implantation, with the acquisition of a receptive phenotype in the uterine epithelium, an initial tenuous attachment of embryonic trophectoderm initiates reorganisation of epithelial polarity to enable stable embryo attachment and the differentiation of invasive trophoblasts. In this Cell Science at a Glance article, we describe cellular and molecular events during the epithelial phase of implantation in rodent, drawing on morphological studies both in vivo and in vitro, and genetic models. Evidence is emerging for a repertoire of transcription factors downstream of the master steroidal regulators estrogen and progesterone that coordinate alterations in epithelial polarity, delivery of signals to the stroma and epithelial cell death or displacement. We discuss what is known of the cell interactions that occur during implantation, before considering specific adhesion molecules. We compare the rodent data with our much more limited knowledge of the human system, where direct mechanistic evidence is hard to obtain. In the accompanying poster, we represent the embryo-epithelium interactions in humans and laboratory rodents, highlighting similarities and differences, as well as depict some of the key cell biological events that enable interstitial implantation to occur. © 2017. Published by The Company of Biologists Ltd.

An Efficient Method for Generation of Transgenic Rats Avoiding Embryo Manipulation

Directory of Open Access Journals (Sweden)

Bhola Shankar Pradhan

2016-01-01

Full Text Available Although rats are preferred over mice as an animal model, transgenic animals are generated predominantly using mouse embryos. There are limitations in the generation of transgenic rat by embryo manipulation. Unlike mouse embryos, most of the rat embryos do not survive after male pronuclear DNA injection which reduces the efficiency of generation of transgenic rat by this method. More importantly, this method requires hundreds of eggs collected by killing several females for insertion of transgene to generate transgenic rat. To this end, we developed a noninvasive and deathless technique for generation of transgenic rats by integrating transgene into the genome of the spermatogonial cells by testicular injection of DNA followed by electroporation. After standardization of this technique using EGFP as a transgene, a transgenic disease model displaying alpha thalassemia was successfully generated using rats. This efficient method will ease the generation of transgenic rats without killing the lives of rats while simultaneously reducing the number of rats used for generation of transgenic animal.

Antigen presenting cells costimulatory signaling during pre-implantation pregnancy 

Directory of Open Access Journals (Sweden)

Anna Sławek

2012-09-01

Full Text Available  Success of pregnancy depends on many factors. Three phenomena inducing immune tolerance against semi-allogeneic conceptus may play a crucial role in the pre-implantation period of pregnancy: influence of sex hormones in sex cycle, presence of oocyte or embryo and the presence of semen in the female reproductive tract. On the other hand dendritic cells are the most effective antigen-presenting cells in regulation of immune phenomena and also are considered as potent participants in inducing immune tolerance in the pregnancy. They communicate with T cells in cell contact-dependent manner or via cytokines. During cell-cell contacts, costimulatory molecules play a key role and their expression is often dependent on cytokines milieu. Both costimulatory molecules and cytokines influence generation of T regulatory cells. Interactions of these molecules are closely related. In this paper we would like to pay attention to the importance of antigen presenting cells costimulatory potency in immune regulation during a pre-implantation period of pregnancy.

De novo DNA methylation during monkey pre-implantation embryogenesis.

Science.gov (United States)

Gao, Fei; Niu, Yuyu; Sun, Yi Eve; Lu, Hanlin; Chen, Yongchang; Li, Siguang; Kang, Yu; Luo, Yuping; Si, Chenyang; Yu, Juehua; Li, Chang; Sun, Nianqin; Si, Wei; Wang, Hong; Ji, Weizhi; Tan, Tao

2017-04-01

Critical epigenetic regulation of primate embryogenesis entails DNA methylome changes. Here we report genome-wide composition, patterning, and stage-specific dynamics of DNA methylation in pre-implantation rhesus monkey embryos as well as male and female gametes studied using an optimized tagmentation-based whole-genome bisulfite sequencing method. We show that upon fertilization, both paternal and maternal genomes undergo active DNA demethylation, and genome-wide de novo DNA methylation is also initiated in the same period. By the 8-cell stage, remethylation becomes more pronounced than demethylation, resulting in an increase in global DNA methylation. Promoters of genes associated with oxidative phosphorylation are preferentially remethylated at the 8-cell stage, suggesting that this mode of energy metabolism may not be favored. Unlike in rodents, X chromosome inactivation is not observed during monkey pre-implantation development. Our study provides the first comprehensive illustration of the 'wax and wane' phases of DNA methylation dynamics. Most importantly, our DNA methyltransferase loss-of-function analysis indicates that DNA methylation influences early monkey embryogenesis.

Effects of sphingosine-1-phosphate on gene expression of two cell mouse embryos induced by C2-Ceramide

Directory of Open Access Journals (Sweden)

Xujing Geng

2014-06-01

Conclusions: This study provides a map of genes in the pre-implantation two cell mouse embryo. Further investigation based on these data will provide a better understanding of the effects of S1P on the pre-implantation embryos in other mammalian species, especially human.

Lethality of radioisotopes in early mouse embryos

International Nuclear Information System (INIS)

Macqueen, H.A.

1979-01-01

The development of pre-implantation mouse embryos was found to be prevented by exposure of the embryos to [ 35 S]methionine, but not to [ 3 H]methionine. Such embryos have also been shown to be highly sensitive to [ 3 H]thymidine. These observations are discussed with reference to the path lengths and energies of electrons emitted from the different radioisotopes. (author)

Developmental defects and genomic instability after x-irradiation of wild-type and genetically modified mouse pre-implantation and early post-implantation embryos

International Nuclear Information System (INIS)

Jacquet, P

2012-01-01

Results obtained from the end of the 1950s suggested that ionizing radiation could induce foetal malformations in some mouse strains when administered during early pre-implantation stages. Starting in 1989, data obtained in Germany also showed that radiation exposure during that period could lead to a genomic instability in the surviving foetuses. Furthermore, the same group reported that both malformations and genomic instability could be transmitted to the next generation foetuses after exposure of zygotes to relatively high doses of radiation. As such results were of concern for radiation protection, we investigated this in more detail during recent years, using mice with varying genetic backgrounds including mice heterozygous for mutations involved in important cellular processes like DNA repair, cell cycle regulation or apoptosis. The main parameters which were investigated included morphological development, genomic instability and gene expression in the irradiated embryos or their own progeny. The aim of this review is to critically reassess the results obtained in that field in the different laboratories and to try to draw general conclusions on the risks of developmental defects and genomic instability from an exposure of early embryos to moderate doses of ionizing radiation. Altogether and in the range of doses normally used in diagnostic radiology, the risk of induction of embryonic death and of congenital malformation following the irradiation of a newly fertilised egg is certainly very low when compared to the ‘spontaneous’ risks for such effects. Similarly, the risk of radiation induction of a genomic instability under such circumstances seems to be very small. However, this is not a reason to not apply some precaution principles when possible. One way of doing this is to restrict the use of higher dose examinations on all potentially pregnant women to the first ten days of their menstrual cycle when conception is very unlikely to have occurred

Proteome Changes in Maize Embryo (Zea mays L) Induced by Ion Beam Implantation Treatment

International Nuclear Information System (INIS)

Li Yongliang; Qin Guangyong; Huo Yuping; Tian Shuangqi; Tang Jihua

2009-01-01

Low energy ion beam implantation was applied to the maize (Zea mays L) embryo proteome using two-dimensional gel electrophoresis. Protein profile analysis detected more than 1100 protein spots, 72 of which were determined to be expressed differently in the treated and control (not exposed to ion beam implantation) embryos. Of the 72 protein spots, 53 were up-regulated in the control and 19 were more abundantly expressed in the ion beam-treated embryos. The spots of up- or down-regulated proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Among the identified proteins, 11 were up-regulated in the treated embryos. Four of these up-regulated proteins were antioxidant molecules, three were related to stress response, two to sugar metabolism and two were associated with heat shock response. Of the five proteins up-regulated in the control embryos, three were functionally related to carbohydrate metabolism; the functions of the remaining two proteins were unknown. The data collected during this study indicate that treatment of maize embryos with low energy ion beam implantation induces changes in stress tolerance enzymes/proteins, possibly as a result of alterations in metabolism. (ion beam bioengineering)

Proteome Changes in Maize Embryo (Zea mays L) Induced by Ion Beam Implantation Treatment

Science.gov (United States)

Li, Yongliang; Tang, Jihua; Qin, Guangyong; Huo, Yuping; Tian, Shuangqi

2009-08-01

Low energy ion beam implantation was applied to the maize (Zea mays L) embryo proteome using two-dimensional gel electrophoresis. Protein profile analysis detected more than 1100 protein spots, 72 of which were determined to be expressed differently in the treated and control (not exposed to ion beam implantation) embryos. Of the 72 protein spots, 53 were up-regulated in the control and 19 were more abundantly expressed in the ion beam-treated embryos. The spots of up- or down-regulated proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Among the identified proteins, 11 were up-regulated in the treated embryos. Four of these up-regulated proteins were antioxidant molecules, three were related to stress response, two to sugar metabolism and two were associated with heat shock response. Of the five proteins up-regulated in the control embryos, three were functionally related to carbohydrate metabolism; the functions of the remaining two proteins were unknown. The data collected during this study indicate that treatment of maize embryos with low energy ion beam implantation induces changes in stress tolerance enzymes/proteins, possibly as a result of alterations in metabolism.

Three-dimensional localisation of NANOG, OCT4, and E-CADHERIN in porcine pre- and peri-implantation embryos

DEFF Research Database (Denmark)

Wolf, Xenia Asbæk; Serup, Palle; Hyttel, Poul

2011-01-01

. The expression of NANOG differed remarkably from that reported in other species. NANOG was not detected in the inner cell mass of hatched porcine blastocysts, but later appeared in the epiblast and hypoblast of spherical blastocysts where Rauber's layer had disintegrated. In pre-gastrulating, filamentous embryos...

Sexing bovine pre-implantation embryos using the polymerase ...

African Journals Online (AJOL)

Yomi

2012-03-06

Mar 6, 2012 ... with pregnancy follow-up to October 2008. Hum. Reprod. 25(11):. 2685-2707. Harper JC, Sengupta SB (2012) Preimplantation genetic diagnosis: State of the ART 2011. Hum. Genet. 131(2): 175-186. Hasler JF (2003). The current status and future of commercial embryo transfer in cattle. Anim. Reprod. Sci.

Uterine responses to early pre-attachment embryos in the domestic dog and comparisons with other domestic animal species.

Science.gov (United States)

Graubner, Felix R; Gram, Aykut; Kautz, Ewa; Bauersachs, Stefan; Aslan, Selim; Agaoglu, Ali R; Boos, Alois; Kowalewski, Mariusz P

2017-08-01

In the dog, there is no luteolysis in the absence of pregnancy. Thus, this species lacks any anti-luteolytic endocrine signal as found in other species that modulate uterine function during the critical period of pregnancy establishment. Nevertheless, in the dog an embryo-maternal communication must occur in order to prevent rejection of embryos. Based on this hypothesis, we performed microarray analysis of canine uterine samples collected during pre-attachment phase (days 10-12) and in corresponding non-pregnant controls, in order to elucidate the embryo attachment signal. An additional goal was to identify differences in uterine responses to pre-attachment embryos between dogs and other mammalian species exhibiting different reproductive patterns with regard to luteolysis, implantation, and preparation for placentation. Therefore, the canine microarray data were compared with gene sets from pigs, cattle, horses, and humans. We found 412 genes differentially regulated between the two experimental groups. The functional terms most strongly enriched in response to pre-attachment embryos related to extracellular matrix function and remodeling, and to immune and inflammatory responses. Several candidate genes were validated by semi-quantitative PCR. When compared with other species, best matches were found with human and equine counterparts. Especially for the pig, the majority of overlapping genes showed opposite expression patterns. Interestingly, 1926 genes did not pair with any of the other gene sets. Using a microarray approach, we report the uterine changes in the dog driven by the presence of embryos and compare these results with datasets from other mammalian species, finding common-, contrary-, and exclusively canine-regulated genes. © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

Comparing rat and rabbit embryo-fetal developmental toxicity ...

Science.gov (United States)

A database of embryo-fetal developmental toxicity (EFDT) studies of 379 pharmaceutical compounds in rat and rabbit was analyzed for species differences based on toxicokinetic parameters of area under the curve (AUC) and maximum concentration (Cmax) at the developmental adverse effect level (dLOAEL). For the vast majority of cases (83% based on AUC of n=283), dLOAELs in rats and rabbits were within the same order of magnitude (less than 10-fold different) when compared based on available data on AUC and Cmax exposures. For 13.5% of the compounds the rabbit was more sensitive and for 3.5% of compounds the rat was more sensitive when compared based on AUC exposures. For 12% of the compounds the rabbit was more sensitive and for 1.3% of compounds the rat was more sensitive based on Cmax exposures. When evaluated based on human equivalent dose (HED) conversion using standard factors, the rat and rabbit were equally sensitive. The relative extent of embryo-fetal toxicity in the presence of maternal toxicity was not different between species. Overall effect severity incidences were distributed similarly in rat and rabbit studies. Individual rat and rabbit strains did not show a different general distribution of systemic exposure LOAELs as compared to all strains combined for each species. There were no apparent species differences in the occurrence of embryo-fetal variations. Based on power of detection and given differences in the nature of developmental effects betwe

Dysregulated LIF-STAT3 pathway is responsible for impaired embryo implantation in a Streptozotocin-induced diabetic mouse model

Directory of Open Access Journals (Sweden)

Tong-Song Wang

2015-07-01

Full Text Available The prevalence of diabetes is increasing worldwide with the trend of patients being young and creating a significant burden on health systems, including reproductive problems, but the effects of diabetes on embryo implantation are still poorly understood. Our study was to examine effects of diabetes on mouse embryo implantation, providing experimental basis for treating diabetes and its complications. Streptozotocin (STZ was applied to induce type 1 diabetes from day 2 of pregnancy or pseudopregnancy in mice. Embryo transfer was used to analyze effects of uterine environment on embryo implantation. Our results revealed that the implantation rate is significantly reduced in diabetic mice compared to controls, and the change of uterine environment is the main reason leading to the decreased implantation rate. Compared to control, the levels of LIF and p-STAT3 are significantly decreased in diabetic mice on day 4 of pregnancy, and serum estrogen level is significantly higher. Estrogen stimulates LIF expression under physiological level, but the excessive estrogen inhibits LIF expression. LIF, progesterone or insulin supplement can rescue embryo implantation in diabetic mice. Our data indicated that the dysregulated LIF-STAT3 pathway caused by the high level of estrogen results in the impaired implantation in diabetic mice, which can be rescued by LIF, progesterone or insulin supplement.

Influence of bone marrow-derived mesenchymal stem cells pre-implantation differentiation approach on periodontal regeneration in vivo.

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Cai, Xinjie; Yang, Fang; Yan, Xiangzhen; Yang, Wanxun; Yu, Na; Oortgiesen, Daniel A W; Wang, Yining; Jansen, John A; Walboomers, X Frank

2015-04-01

The implantation of bone marrow-derived mesenchymal stem cells (MSCs) has previously been shown successful to achieve periodontal regeneration. However, the preferred pre-implantation differentiation strategy (e.g. maintenance of stemness, osteogenic or chondrogenic induction) to obtain optimal periodontal regeneration is still unknown. This in vivo study explored which differentiation approach is most suitable for periodontal regeneration. Mesenchymal stem cells were obtained from Fischer rats and seeded onto poly(lactic-co-glycolic acid)/poly(ɛ-caprolactone) electrospun scaffolds, and then pre-cultured under different in vitro conditions: (i) retention of multilineage differentiation potential; (ii) osteogenic differentiation approach; and (iii) chondrogenic differentiation approach. Subsequently, the cell-scaffold constructs were implanted into experimental periodontal defects of Fischer rats, with empty scaffolds as controls. After 6 weeks of implantation, histomorphometrical analyses were applied to evaluate the regenerated periodontal tissues. The chondrogenic differentiation approach showed regeneration of alveolar bone and ligament tissues. The retention of multilineage differentiation potential supported only ligament regeneration, while the osteogenic differentiation approach boosted alveolar bone regeneration. Chondrogenic differentiation of MSCs before implantation is a useful strategy for regeneration of alveolar bone and periodontal ligament, in the currently used rat model. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Alternation of apoptotic and implanting genes expression of mouse embryos after re-vitrification

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Majidi Gharenaz, Nasrin; Movahedin, Mansoureh; Mazaheri, Zohreh; Pour beiranvand, Shahram

2016-01-01

Background: Nowadays, oocytes and embryos vitrification has become a routine technique. Based on clinical judgment, re-vitrification maybe required. But little is known about re-vitrification impact on genes expression. Objective: The impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study. Materials and Methods: In this experimental study, 8 cell embryos (n=240) were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh (n=80), vitrified at 8 cell stage (n=80), vitrified at 8 cell stage thawed and re-vitrified at compaction stage (n=80). Embryos were vitrified by using cryolock, (open system) described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts. Results: Our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos (p=0.03). In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos (p=0.004), however expression of Bax and Bcl-2 (apoptotic) genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos (p=0.003), but it was similar between re-vitrified and vitrified embryos. Conclusion: Re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage. PMID:27679826

Influences of serum from ozone-exposed pregnant rats in an in vitro model of implantation

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In our previous studies, ozone (O3) exposure during implantation [gestational day (GD) 5 and 6)] in rats resulted in intrauterine growth restriction (IUGR), suggesting impairment of implantation with exposure. The aim of this study was to (1) determine if serum collected from pre...

Effects of melatonin administration on embryo implantation and offspring growth in mice under different schedules of photoperiodic exposure.

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Zhang, Lu; Zhang, Zhenzhen; Wang, Feng; Tian, Xiuzhi; Ji, Pengyun; Liu, Guoshi

2017-10-02

Embryo implantation is crucial for animal reproduction. Unsuccessful embryo implantation leads to pregnancy failure, especially in human-assisted conception. Environmental factors have a profound impact on embryo implantation. Because people are being exposed to more light at night, the influence of long-term light exposure on embryo implantation should be explored. The effects of long photoperiodic exposure and melatonin on embryo implantation and offspring growth were examined. Long photoperiodic exposure (18:6 h light:dark) was selected to resemble light pollution. Melatonin (10 -2 , 10 -3 , 10 -4 , 10 -5  M) was added to the drinking water of mice starting at Day 1 (vaginal plugs) until delivery. Melatonin treatment (10 -4 ,10 -5  M) significantly increased litter sizes compared to untreated controls (12.9 ± 0.40 and 12.2 ± 1.01 vs. 11.5 ± 0.43; P melatonin (10 -4  M) was selected for further investigation. No remarkable differences were found between melatonin-treated mice and controls in terms of the pups' birth weights, weaning survival rates, and weaning weights. Long photoperiodic exposure significantly reduced the number of implantation sites in treated mice compared to controls (light/dark, 12/12 h), and melatonin rescued this negative effect. Mechanistic studies revealed that melatonin enhanced the serum 17β-estradiol (E 2 ) levels in the pregnant mice and upregulated the expression of the receptors MT1 and MT2 and p53 in uterine tissue. All of these factors may contribute to the beneficial effects of melatonin on embryo implantation in mice. Melatonin treatment was associated with beneficial effects in pregnant mice, especially those subjected to long photoperiodic exposure. This was achieved by enhanced embryo implantation. At the molecular level, melatonin administration probably increases the E 2 level during pregnancy and upregulates p53 expression by activating MT1/2 in the uterus. All of the changes may improve the

Alternation of apoptotic and implanting genes expression of mouse embryos after re-vitrification

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Nasrin Majidi Gharenaz

2016-08-01

Full Text Available Background: Nowadays, oocytes and embryos vitrification has become a routine technique. Based on clinical judgment, re-vitrification maybe required. But little is known about re-vitrification impact on genes expression. Objective: The impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study. Materials and Methods: In this experimental study, 8 cell embryos (n=240 were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh (n=80, vitrified at 8 cell stage (n=80, vitrified at 8 cell stage thawed and re-vitrified at compaction stage (n=80. Embryos were vitrified by using cryolock, (open system described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts. Results: Our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos (p=0.03. In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos (p=0.004, however expression of Bax and Bcl-2 (apoptotic genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos (p=0.003, but it was similar between re-vitrified and vitrified embryos. Conclusion: Re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage

Local activation of uterine Toll-like receptor 2 and 2/6 decreases embryo implantation and affects uterine receptivity in mice.

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Sanchez-Lopez, Javier Arturo; Caballero, Ignacio; Montazeri, Mehrnaz; Maslehat, Nasim; Elliott, Sarah; Fernandez-Gonzalez, Raul; Calle, Alexandra; Gutierrez-Adan, Alfonso; Fazeli, Alireza

2014-04-01

Embryo implantation is a complex interaction between maternal endometrium and embryonic structures. Failure to implant is highly recurrent and impossible to diagnose. Inflammation and infections in the female reproductive tract are common causes of infertility, embryo loss, and preterm labor. The current work describes how the activation of endometrial Toll-like receptor (TLR) 2 and 2/6 reduces embryo implantation chances. We developed a morphometric index to evaluate the effects of the TLR 2/6 activation along the uterine horn (UH). TLR 2/6 ligation reduced the endometrial myometrial and glandular indexes and increased the luminal index. Furthermore, TLR 2/6 activation increased the proinflammatory cytokines such as interleukin (IL)-1beta and monocyte chemotactic protein (MCP)-1 in UH lavages in the preimplantation day and IL-1 receptor antagonist in the implantation day. The engagement of TLR 2/6 with its ligand in the UH during embryo transfer severely affected the rate of embryonic implantation (45.00% ± 6.49% vs. 16.69% ± 5.01%, P embryo implantation process was verified using an in vitro model of human embryo implantation where trophoblast spheroids failed to adhere to a monolayer of TLR 2- and TLR 2/6-activated endometrial cells. The inhibition of TLR receptors 2 and 6 in the presence of their specific ligands restored the ability of the spheroids to bind to the endometrial cells. In conclusion, the activation of the innate immune system in the uterus at the time of implantation interfered with the endometrial receptivity and reduced the chances of implantation success.

No evidence of association of MUC-1 genetic polymorphism with embryo implantation failure

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D.B. Dentillo

2007-06-01

Full Text Available Pregnancy loss can be caused by several factors involved in human reproduction. Although up to 50% of cases remain unexplained, it has been postulated that the major cause of failed pregnancy is an error of embryo implantation. Transmembrane mucin-1 (MUC-1 is a glycoprotein expressed on the endometrial cell surface which acts as a barrier to implantation. The gene that codes for this molecule is composed of a polymorphic tandem repeat of 60 nucleotides. Our objective was to determine if MUC-1 genetic polymorphism is associated with implantation failure in patients with a history of recurrent abortion. The study was conducted on 10 women aged 25 to 35 years with no history of successful pregnancy and with a diagnosis of infertility. The control group consisted of 32 patients aged 25 to 35 years who had delivered at least two full-term live children and who had no history of abortions or fetal losses. MUC-1 amplicons were obtained by PCR and observed on agarose and polyacrylamide gel after electrophoresis. Statistical analysis showed no significant difference in the number of MUC-1 variable number of tandem repeats between these groups (P > 0.05. Our results suggest that there is no effect of the polymorphic MUC-1 sequence on the implantation failure. However, the data do not exclude MUC-1 relevance during embryo implantation. The process is related to several associated factors such as the mechanisms of gene expression in the uterus, specific MUC-1 post-translational modifications and appropriate interactions with other molecules during embryo implantation.

The rat whole embryo culture assay using the Dysmorphology Score system.

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Zhang, Cindy; Panzica-Kelly, Julie; Augustine-Rauch, Karen

2013-01-01

The rat whole embryo culture (WEC) system has been used extensively for characterizing teratogenic properties of test chemicals. In this chapter, we describe the methodology for culturing rat embryos as well as a new morphological score system, the Dysmorphology Score (DMS) system for assessing morphology of mid gestation (gestational day 11) rat embryos. In contrast to the developmental stage focused scoring associated with the Brown and Fabro score system, this new score system assesses the respective degree of severity of dysmorphology, which delineates normal from abnormal morphology of specific embryonic structures and organ systems. This score system generates an approach that allows rapid identification and quantification of adverse developmental findings, making it conducive for characterization of compounds for teratogenic properties and screening activities.

Effect of Cumulus cell co-culture and Protein Supplement on Success of in vitro Fertilization and Development of Pre-implanted Embryos in mice

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Muhammad-Baqir M-R. Fakhrildin

2005-06-01

Full Text Available Successful oocyte fertilization and normal embryonic development of mice were considered the most important diagnostic criteria for the safety of materials and tools used for human in vitro fertilization and embryo transfer (IVF-ET. Therefore, we studied the influence of cumulus cells co-culture and protein supplement within culture medium on percentages of in vitro fertilization (IVF and normal development of early stages of mouse embryo later. Oocytes were collected and treated with hyaluronidase to remove cumulus cells. Oocytes were divided into four groups namely: Group-1: Oocytes incubated within modified Earl’s medium (MEM supplied with 10% inactivated bovine amniotic fluid as a protein source and cumulus cells; Group-2: Oocytes incubated with MEM supplied with cumulus cells only; Group-3: Oocytes incubated with MEM supplied with 10% inactivated bovine amniotic fluid only; and Group-4: Oocytes  incubated with MEM free of both protein source and cumulus cells. For IVF, 5-6 oocytes were incubated with active spermatozoa under paraffin oil for 18-20 hours at 37° oC in 5% CO2. Percentages of IVF and embryonic development were then recorded. Best results for IVF and normal embryonic development were achieved from oocytes of Group-1 when compared to the other groups. As compared to Group-1, the percentage of IVF for Group-2 and Group-3 were decreased insignificantly and significantly (P<0.002, respectively. Significant (P<0.01 reduction in the percentages of IVF and normal embryonic development were reported in Group-4 as compared to Group-1. Therefore, it was concluded that the presence of cumulus cells co-culture and bovine amniotic fluid as a protein source within culture medium may have an important role on the fertilizing capacity of spermatozoa and oocytes and normal development of pre-implanted mouse embryo later.

Influence of the radiation (Co60) in pre-implants rabbit embryos: effect on atypic mitotic index and embryo pole development

International Nuclear Information System (INIS)

Approbato, Mario S.; Oliveira Moura, Katia K.V. de; Souza Florencio, Rodopiano de; Garcia, Ricardo; Faria, Renato S.; Benedetti, Leonardo N.; Goulart, Flamarion B.

1995-01-01

We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), at three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses: five c Gy and ten c Gy. Six rabbits (36 blastocysts) were used as controls. the matching instant was the zero hour. Exactly six days after (± 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. the embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryo parameters were studied: embryo pole development; percentage of abnormal mitotic figures. irradiation time was associated with lower scores of embryo pole development, but not with irradiation dose. There were no gross abnormalities of embryo pole. The abnormal mitotic cells was affected both by the time and dose of irradiation. (author)

Influence of irradiation (Co60) in pre-implant rabbits embryos: effect on blastocyst diameters and embryos smaller than 2 mm

International Nuclear Information System (INIS)

Approbato, Mario S.; Oliveira Moura, Katia K.V. de; Souza Florencio, Rodopiano de; Cunha Junior, Carlos; Garcia, Ricardo; Faria, Renato S.; Benedetti, Leonardo N.; Goulart, Flamarion B.

1995-01-01

We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), in three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses, 5 c Gy and 10 c Gy. Six rabbits (36 blastocysts) were used as controls. The matching instant was the zero hour. Exactly six days after (± 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. The embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryos parameters were studied: diameter growth; percentage of embryos smaller than 2 mm. We observed that only the irradiation time influenced the blastocysts diameter (no irradiation dose). There was no relation between percentage of embryos smaller than 2 mm and the irradiation. (author)

MicroRNA-200a locally attenuates progesterone signaling in the cervix, preventing embryo implantation.

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Haraguchi, Hirofumi; Saito-Fujita, Tomoko; Hirota, Yasushi; Egashira, Mahiro; Matsumoto, Leona; Matsuo, Mitsunori; Hiraoka, Takehiro; Koga, Kaori; Yamauchi, Naoko; Fukayama, Masashi; Bartos, Amanda; Cha, Jeeyeon; Dey, Sudhansu K; Fujii, Tomoyuki; Osuga, Yutaka

2014-07-01

Although cervical pregnancy and placenta previa, in which the embryo and placenta embed in or adjacent to the cervix, are life-threatening complications that result in massive bleeding and poor pregnancy outcomes in women, the incidence of these aberrant conditions is uncommon. We hypothesized that a local molecular mechanism is normally in place to prevent embryo implantation in the cervix. The ovarian hormones progesterone (P(4)) and estrogen differentially direct differentiation and proliferation of endometrial cells, which confers the receptive state for implantation: P(4) dominance causes differentiation of the luminal epithelium but increases stromal cell proliferation in preparation of the uterus for implantation. In search for the cause of cervical nonresponsiveness to implantation, we found that the statuses of cell proliferation and differentiation between the uterus and cervix during early pregnancy are remarkably disparate under identical endocrine milieu in both mice and humans. We also found that cervical levels of progesterone receptor (PR) protein are low compared with uterine levels during this period, and the low PR protein levels are attributed to elevated levels of microRNA(miR)-200a in the cervix. These changes were associated with up-regulation of the P(4)-metabolizing enzyme 20α-hydroxysteroid dehydrogenase (200α-HSD) and down-regulation of its transcriptional repressor signal transducer and activator of transcription 5 in the cervix. The results provide evidence that elevated levels of miR-200a lead to down-regulation of P(4)-PR signaling and up-regulation of (200α-HSD) in the cervix, rendering it nonresponsive to implantation. These findings may point toward not only the physiological but also the pathological basis of the cervical milieu in embryo implantation.

Embryonic mortality and intrauterine growth retardation (IUGR) associated with placental alterations in pregnant rats treated with methyl methanesulfonate (MMS) at the peri-implantation stage.

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Yokoi, Ryohei; Hayashi, Morimichi; Tamura, Toru; Kobayashi, Kazuo; Kuroda, Junji; Kusama, Hiroshi; Kagami, Hiroshi; Ono, Tamao

2008-12-01

Embryonic mortality and intrauterine growth retardation (IUGR) are induced by exposure of rodents to xenobiotic agents during the pregastrulation period of development. We examined the time course of the effects of methyl methanesulfonate (MMS), an alkylating agent, on conceptus development in order to clarify the relative roles of the embryo and the placenta in their induction. Pregnant rats were treated orally with a single dose of MMS (200 mg/kg) in the morning of gestation day (GD) 6 (peri-implantation stage). Embryonic mortality was increased on GD12 and thereafter by MMS treatment, with newly dead embryos showing placental hypoplasia at GD12. Embryo or fetal weight was also smaller for MMS-treated dams than for control dams from GD14 to GD20. The labyrinth zone and junctional zone (JZ) of the placenta were thinner in MMS-treated rats from GD12 to GD17 and from GD12 to GD20 (except for GD17), respectively. Furthermore, MMS-treated dams showed a smaller number of glycogen cells in the JZ on GD14. In contrast, the placental glycogen concentration was higher and the expression of glucose transporter 1 in the JZ remained at GD20. These results indicate that exposure of pregnant rats to MMS at the peri-implantation stage of embryogenesis affects placental development and growth. The placental impairment induced by MMS was likely responsible for the embryonic death observed 6 days after exposure of dams to this agent as well as for the IUGR of surviving embryos or fetuses throughout the gestation period.

No specific gene expression signature in human granulosa and cumulus cells for prediction of oocyte fertilisation and embryo implantation.

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Tanja Burnik Papler

Full Text Available In human IVF procedures objective and reliable biomarkers of oocyte and embryo quality are needed in order to increase the use of single embryo transfer (SET and thus prevent multiple pregnancies. During folliculogenesis there is an intense bi-directional communication between oocyte and follicular cells. For this reason gene expression profile of follicular cells could be an important indicator and biomarker of oocyte and embryo quality. The objective of this study was to identify gene expression signature(s in human granulosa (GC and cumulus (CC cells predictive of successful embryo implantation and oocyte fertilization. Forty-one patients were included in the study and individual GC and CC samples were collected; oocytes were cultivated separately, allowing a correlation with IVF outcome and elective SET was performed. Gene expression analysis was performed using microarrays, followed by a quantitative real-time PCR validation. After statistical analysis of microarray data, there were no significantly differentially expressed genes (FDR<0,05 between non-fertilized and fertilized oocytes and non-implanted and implanted embryos in either of the cell type. Furthermore, the results of quantitative real-time PCR were in consent with microarray data as there were no significant differences in gene expression of genes selected for validation. In conclusion, we did not find biomarkers for prediction of oocyte fertilization and embryo implantation in IVF procedures in the present study.

Effect of osteoporosis on fixation of osseointegrated implants in rats.

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Li, Yunfeng; He, Sheng; Hua, Yunwei; Hu, Jing

2017-11-01

The effect of osteoporosis on implant osseointegration has been widely investigated, whereas osteoporosis may also newly occur in patient with previously osseointegrated implant. This study was designed to investigate the effect of osteoporosis on implant fixation in rats after successful osseointegration had been obtained. Seventy female Sprague-Dawley rats were included, and each animal received two titanium implants in the distal metaphysis of femur bilaterally. Eight weeks later, ten rats were sacrificed to confirm the establishment of implant osseointegration. All left rats were randomly subjected to bilateral ovariectomy (OVX) or sham operation. Three, six, and twelve weeks later, implant osseointegration, peri-implant bone tissue, and biomechanical properties of implant were analyzed. Right femurs with implants were used for micro-CT and histological analysis, and left femurs with implants were used for biomechanical test. Micro-CT, histology, and biomechanical test confirmed the destructive effect of OVX on previously osseointegrated implant in rats; when compared to sham-operated rats, peri-implant bone volume, trabecular architecture, bone-to-implant contact ratio, as well as biomechanical parameters decreased progressively within 12 weeks. Results also indicated that the effect of OVX on undisturbed bone (proximal tibiae) was much stronger than that on peri-implant bone. Osteoporosis produced a progressive negative effect on previously osseointegrated implant in distal femora of rats during 12 weeks. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2426-2432, 2017. © 2016 Wiley Periodicals, Inc.

Comparison of rat and rabbit embryo-fetal developmental ...

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Regulatory non-clinical safety testing of human pharmaceutical compounds typically requires embryo fetal developmental toxicity (EFDT) testing in two species, (one rodent and one non-rodent, usually the rat and the rabbit). The question has been raised whether under some conditions EFDT testing could be limited to one species, or whether the need for testing in a second species could be decided on a case by case basis. As part of an RIVM/CBG-MEB/HESI/US EPA consortium initiative, we built and queried a database of 379 EFDT studies conducted for marketed and non-marketed pharmaceutical compounds. The animal models (rat and rabbit) were assessed for their potential for adverse developmental and maternal outcomes. The database was analyzed for the prevalence of EFDT incidence and the nature and severity of adverse findings in the two species. Some manifestation of EFDT in either one or both species (rat and rabbit) was demonstrated for 282 compounds (74%), and EFDT was detected in only one species (rat or rabbit) in almost a third (31%, 118 compounds), with approximately 58% rat and 42% rabbit studies identifying an EFDT signal among the 379 compounds tested. For 24 compounds (6%), fetal malformations were observed in one species (rat or rabbit) in the absence of any EFDT in the second species. In general, growth retardation, fetal variations, and malformations were more prominent in the rat, whereas embryo-fetal death was observed more often in the rabbit. Discor

Neuronal damage in chick and rat embryos following X-irradiation

International Nuclear Information System (INIS)

Schneider, B.F.; Norton, S.

1980-01-01

Exposure of rat and chick embryos to X-irradiation at the time of development of neurons at the telencephalic-diencephalic border results in prolonged damage to neurons in this area as measured by neuronal nuclear size. A dose of 100 rads to the seven-day-old chick embryo has about the same effect as 125 rads to the 15-day-old rat fetus. The nuclear volume of large, multipolar neurons in the chick paleostriatum primitivum and the rat lateral preoptic area are reduced from 10 to 15%. Larger doses of X-irradiation to the chick (150 and 200 rads) cause progressively greater reductions in nuclear size. The large neurons which were measured in the rat and chick are morphologically similar in the two species. Both contain cytoplasmic acetylcholinesterase and have several branched, spiny dendritic processes. The similarity of response of chick and rat neurons to X-irradiation diminishes the significance of maternal factors as the cause of the effects of fetal irradiation in these experiments

Dose-response analysis of phthalate effects on gene expression in rat whole embryo culture

NARCIS (Netherlands)

Robinson, J.F.; Verhoef, A.; van Beelen, V.A.; Pennings, J.L.A.; Piersma, A.H.|info:eu-repo/dai/nl/071276947

2012-01-01

The rat postimplantation whole embryo culture (WEC) model serves as a potential screening tool for developmental toxicity. In this model, cultured rat embryos are exposed during early embryogenesis and evaluated for morphological effects. The integration of molecular-based markers may lead to

Three-Dimensional High-Frequency Ultrasonography for Early Detection and Characterization of Embryo Implantation Site Development in the Mouse.

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Mary C Peavey

Full Text Available Ultrasonography is a powerful tool to non-invasively monitor in real time the development of the human fetus in utero. Although genetically engineered mice have served as valuable in vivo models to study both embryo implantation and pregnancy progression, such studies usually require sacrifice of parous mice for subsequent phenotypic analysis. To address this issue, we used three-dimensional (3-D reconstruction in silico of high-frequency ultrasound (HFUS imaging data for early detection and characterization of murine embryo implantation sites and their development in utero. With HFUS imaging followed by 3-D reconstruction, we were able to precisely quantify embryo implantation site number and embryonic developmental progression in pregnant C57BL6J/129S mice from as early as 5.5 days post coitus (d.p.c. through to 9.5 d.p.c. using a VisualSonics Vevo 2100 (MS550S transducer. In addition to measurements of implantation site number, location, volume and spacing, embryo viability via cardiac activity monitoring was also achieved. A total of 12 dams were imaged with HFUS with approximately 100 embryos examined per embryonic day. For the post-implantation period (5.5 to 8.5 d.p.c., 3-D reconstruction of the gravid uterus in mesh or solid overlay format enabled visual representation in silico of implantation site location, number, spacing distances, and site volume within each uterine horn. Therefore, this short technical report describes the feasibility of using 3-D HFUS imaging for early detection and analysis of post-implantation events in the pregnant mouse with the ability to longitudinally monitor the development of these early pregnancy events in a non-invasive manner. As genetically engineered mice continue to be used to characterize female reproductive phenotypes, we believe this reliable and non-invasive method to detect, quantify, and characterize early implantation events will prove to be an invaluable investigative tool for the study of

Effects of elevated ambient temperature on embryo implantation in rats

African Journals Online (AJOL)

Yomi

2012-03-22

Mar 22, 2012 ... ambient temperature leads to a delayed implantation and reduced number of implantation sites in. Sprague ... rates decrease after exposure to stress. One of the ..... implantation initiation time, support the previous findings.

Effect of organically bound tritium (OBT) on pre-implantation mouse embryos in vitro

International Nuclear Information System (INIS)

Yamada, Takeshi; Ohyama, Harumi

1989-01-01

Effect of organically bound tritium (OBT), such as tritiated thymidine and tritium-labeled amino acids, on mouse preimplantation embryos was examined in vitro. Mouse zygotes fertilized in vitro (BC3F 1 eggs x ICR sperm) were cultured in the media containing OBT in various concentrations up to the blastocyst stage. The LD 50 in terms of tritium concentrations in the culture medium were determined by measuring tritium concentrations in the medium to inhibit 50 % of embryos to form blastocyst in vitro. Tritium activities in the embryos were measured at various times during culture of embryos at LD 50 concentration in order to estimate absorbed radiation dose in embryonic cells. The LD 50 values obtained indicate that OBT could inhibit the embryonic development 1000 times more effectively that tritiated water (HTO). However, differences in LD 50 values in terms of absorbed radiation dose between OBT and HTO is not so essential, and might be explained by localized spatial distribution of OBT within the cell. (author)

Effect of alpha-lipoic acid on endometrial implants in an experimental rat model.

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Pınar, Neslihan; Soylu Karapınar, Oya; Özcan, Oğuzhan; Özgür, Tümay; Bayraktar, Suphi

2017-10-01

To investigate the antioxidant and anti-inflammatory effects of alpha-lipoic acid (ALA) in the treatment of endometriosis in an experimental rat model by evaluating biochemical and histopathologic parameters. Experimental endometriosis was induced by the peritoneal implantation of autologous endometrial tissue. The rats were randomly divided into two groups with eight rats each. Group I was intraperitoneally administered ALA 100 mg/kg/day for 14 days. Group II was intraperitoneally administered saline solution at the same dosage and over the same period. Endometrial implant volume was measured in both groups both pre- and post-treatment. Tumor necrosis factor alpha (TNF-α) was measured in peritoneal fluid. Total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) were assessed in serum. The implants were histopathologically evaluated. In the ALA group, the serum TOS and OSI levels, the endometrial implant volumes, the TNF-α levels in serum and peritoneal fluid, and the histopathologic scores were significantly lower compared to the control group (P < 0.05). Alpha-lipoic acid may have a therapeutic potential in the treatment of endometriosis due to its antioxidant and anti-inflammatory effects. © 2017 Société Française de Pharmacologie et de Thérapeutique.

Influences of granulocyte growth factor in uterine perfusion on pregnancy outcome of patients with failure of embryo implantation for unknown reason.

Science.gov (United States)

He, Jun; Liu, Juan; Zhou, Hua; Chen, Chao Jun

2016-11-01

To investigate the influence of granulocyte growth factor in uterine perfusion on the pregnancy outcome of patients with failure of embryo implantation for unknown reason. Then, 68 patients with failure of embryo implantation for unknown reason were enrolled in our hospital from November 2013 to February 2015, which were divided into observation group and control group by random (34 patients in each group). Patients in observation group received basic treatment for granulocyte growth factor in uterine perfusion on the next day, while patients in control group received basic treatment with placebo. Then, endometrial preparation, adverse reaction and pregnancy outcome of patients were compared between the two groups. Comparing the endometrial preparation and average endometrial thickness of patients in control group (9.87±2.12) with those in observation group [(9.87±2.12), there is no significant difference (Pfactor, patients with failure of embryo implantation can effectively improve clinical pregnancy rate and embryo implantation rate without severe complication. Therefore, treatment of granlocyte growth factor can improve the pregnancy outcome of patients.

Estimating limits for natural human embryo mortality [version 2; referees: 2 approved

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Gavin E. Jarvis

2016-12-01

Full Text Available Natural human embryonic mortality is generally considered to be high. Values of 70% and higher are widely cited. However, it is difficult to determine accurately owing to an absence of direct data quantifying embryo loss between fertilisation and implantation. The best available data for quantifying pregnancy loss come from three published prospective studies (Wilcox, Zinaman and Wang with daily cycle by cycle monitoring of human chorionic gonadotrophin (hCG in women attempting to conceive. Declining conception rates cycle by cycle in these studies indicate that a proportion of the study participants were sub-fertile. Hence, estimates of fecundability and pre-implantation embryo mortality obtained from the whole study cohort will inevitably be biased. This new re-analysis of aggregate data from these studies confirms the impression that discrete fertile and sub-fertile sub-cohorts were present. The proportion of sub-fertile women in the three studies was estimated as 28.1% (Wilcox, 22.8% (Zinaman and 6.0% (Wang. The probability of conceiving an hCG pregnancy (indicating embryo implantation was, respectively, 43.2%, 38.1% and 46.2% among normally fertile women, and 7.6%, 2.5% and 4.7% among sub-fertile women. Pre-implantation loss is impossible to calculate directly from available data although plausible limits can be estimated. Based on this new analysis and a model for evaluating reproductive success and failure it is proposed that a plausible range for normal human embryo and fetal mortality from fertilisation to birth is 40-60%.

Immune responses to implanted human collagen graft in rats

International Nuclear Information System (INIS)

Quteish, D.; Dolby, A.E.

1991-01-01

Immunity to collagen implants may be mediated by cellular and humoral immune responses. To examine the possibility of such immunological reactivity and crossreactivity to collagen, 39 Sprague-Dawley rats (female, 10 weeks old, approximately 250 g wt) were implanted subcutaneously at thigh sites with crosslinked, freeze-dried human placental type I collagen grafts (4x4x2 mm) which had been irradiated (520 Gray) or left untreated. Blood was obtained by intracardiac sampling prior to implantation or from normal rats, and at various times afterwards when the animals were sacrificed. The sera from these animals were examined for circulating antibodies to human, bovine and rat tail (type I) collagens by enzyme-linked immunosorbent assay (ELISA). Also, the lymphoblastogenic responses of spleen lymphocytes from the irradiated collagen-implanted animals were assessed in culture by measuring thymidine uptake with autologous and normal rat sera in the presence of human bovine type I collagens. Implantation of the irradiated and non-irradiated collagen graft in rats led to a significant increase in the level of circulating antibodies to human collagen. Also antibody to bovine and rat tail collagens was detectable in the animals implanted with irradiated collagen grafts but at a lower level than the human collagen. There was a raised lymphoblastogenic response to both human and bovine collagens. The antibody level and lymphoblastogenesis to the tested collagens gradually decreased towards the end of the post-implantation period. (author)

Maternal expression of the histone demethylase Kdm4a is crucial for pre-implantation development

DEFF Research Database (Denmark)

Sankar, Aditya; Kooistra, Susanne Marije; Gonzalez, Javier Martin

2017-01-01

-methylated lysine 9 and lysine 36 of histone H3 (H3K9me2/me3 and H3K36me2/me3). Here, we report that Kdm4a as a maternal factor plays a key role in embryo survival and is vital for female fertility. Kdm4a−/− female mice ovulate normally with comparable fertilization but poor implantation rates, and cannot support......-deficient oocytes displays a poor intrinsic ability to develop into blastocysts. These embryos cannot compete with healthy embryos for implantation in vivo, highlighting Kdm4a as a maternal effect gene. Thus, our study dissects an important dual role for maternal Kdm4a in determining faithful early embryonic...... healthy transplanted embryos to term. This is due to a role for Kdm4a in uterine function, where its loss causes reduced expression of key genes involved in ion transport, nutrient supply and cytokine signalling, which impact embryo survival. In addition, a significant proportion of Kdm4a...

Cytokine-Like Protein 1(Cytl1: A Potential Molecular Mediator in Embryo Implantation.

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Zhichao Ai

Full Text Available Cytokine-like protein 1 (Cytl1, originally described as a protein expressed in CD34+ cells, was recently identified as a functional secreted protein involved in chondrogenesis and cartilage development. However, our knowledge of Cytl1 is still limited. Here, we determined the Cytl1 expression pattern regulated by ovarian hormones at both the mRNA and protein levels. We found that the endometrial expression of Cytl1 in mice was low before or on the first day of gestation, significantly increased during embryo implantation, and then decreased at the end of implantation. We investigated the effects of Cytl1 on endometrial cell proliferation, and the effects on the secretion of leukemia inhibitory factor (LIF and heparin-binding epidermal growth factor (HB-EGF. We also explored the effect of Cytl1 on endometrial adhesion properties in cell-cell adhesion assays. Our findings demonstrated that Cytl1 is an ovarian hormone-dependent protein expressed in the endometrium that enhances the proliferation of HEC-1-A and RL95-2 cells, stimulates endometrial secretion of LIF and HB-EGF, and enhances the adhesion of HEC-1-A and RL95-2 cells to JAR spheroids. This study suggests that Cytl1 plays an active role in the regulation of embryo implantation.

Pre-incubation of porcine semen reduces the incidence of polyspermy on embryos derived from low quality oocytes

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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.

The Effects of Maternal Hyperthyroidism on Histologic Changes in Parietal Lobe in Rat Embryos

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Fatemeh Mirsafi

2017-05-01

Full Text Available Background Maternal hyperthyroidism causes developmental defects on the nervous system of fetuses. Objectives The present study was designed to study the effects of maternal hyperthyroidism on the development of the parietal lobe in the brain of rat embryos. Methods In this experimental study, thirty Sprague-Dawley rats were randomly divided into three groups. The control group received no injections, the sham group received intraperitoneal injections of distilled water solution containing salt and polysorbate (solvent of levothyroxine, and the experimental group received once-daily, intraperitoneal injections of 0.5 mg/kg levothyroxine for a 10-day period to become hyperthyroid rats. The hyperthyroid rats were then mated, and all pregnant rats were killed on the 20th day of gestation. Fetuses were removed, fixed, and processed for histological procedures. The fetuses were sagitally sectioned at 5 µ thickness and stained with hematoxylin-eosin (H and E technique. The sections were examined using a light microscope and Motic software. Results The results showed no significant difference in the studied variables between the sham and control groups. A significantly increase in body weight and a significant decrease in crown-rump length of embryos was observed in the experimental group when compared to the control group. The mean total thickness of the parietal cortex, ventricular layer, and intermediate layer of embryos showed a significant decrease in the experimental group compared to the control and sham groups. The mean number of cells also showed a significant decrease in the intermediate and ventricular layers in the experimental group compared to the control and sham groups. Conclusions This study showed that maternal hyperthyroidism leads to a reduction in development of the parietal cortex in embryos. Maternal hyperthyroidism can disturb the growth and development of embryos.

Embryo quality and implantation rate in two different culture media: ISM1 versus Universal IVF Medium.

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Xella, Susanna; Marsella, Tiziana; Tagliasacchi, Daniela; Giulini, Simone; La Marca, Antonio; Tirelli, Alessandra; Volpe, Annibale

2010-04-01

To compare the outcome of two different culture media marketed by the MediCult AS Company (Jyllinge, Denmark)-Universal IVF Medium and ISM1 Medium culture-which, in addition to glucose, pyruvate, and energy-providing components, also contain amino acids, nucleotides, vitamins, and cholesterol. Laboratory and retrospective clinical study. University teaching hospital. A total of 726 patients, undergoing IVF-intracytoplasmic sperm injection procedure, comparable in mean age range, oocyte retrieval, and infertility indication, were included in the study. Laboratory quality and standard procedures were maintained unaffected. Oocyte retrieval, different embryo culture media. Embryo quality, ongoing pregnancy, and implantation rate. The frequency of good-quality embryos (79% vs. 74%) and the percentages of ongoing pregnancy (27.5% vs. 18%) and implantation rate (15% vs. 10%) were significantly higher in the group treated with ISM1 Medium rather than Universal IVF Medium. ISM1 Medium culture seems to improve the performance of embryonic growth and development, as well as increasing the percentage of pregnancy. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

9 CFR 85.10 - Interstate movement of swine semen and swine embryos for insemination of or implantation into swine.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Interstate movement of swine semen and... ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS PSEUDORABIES § 85.10 Interstate movement of swine semen and swine embryos for insemination of or implantation into swine. Swine semen and swine embryos moved...

LIF supports primitive endoderm expansion during pre-implantation development

DEFF Research Database (Denmark)

Morgani, Sophie M; Brickman, Joshua M

2015-01-01

Embryonic stem cells (ESCs) are pluripotent cell lines that can be maintained indefinitely in an early developmental state. ESC culture conditions almost always require the cytokine LIF to maintain self-renewal. As ESCs are not homogeneous but contain multiple populations reminiscent...... of the blastocyst, identifying the target cells of LIF is necessary to understand the propagation of pluripotency. We recently found that LIF acts under self-renewing conditions to stimulate the fraction of ESCs that express extraembryonic markers, but has little impact on pluripotent gene expression. Here, we...... derivatives, whereas the increase in PrE is mediated both by an increase in proliferation and inhibition of PrE apoptosis that is normally triggered in embryos with an excess of GATA6(+) cells. Thus, it appears that the relative size of the PrE is determined by the number of LIF-producing cells in the embryo...

Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy

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Bradley, Josephine; Pope, Iestyn; Masia, Francesco; Sanusi, Randa; Langbein, Wolfgang; Borri, Paola

2016-01-01

Mammalian oocytes contain lipid droplets that are a store of fatty acids, whose metabolism plays a substantial role in pre-implantation development. Fluorescent staining has previously been used to image lipid droplets in mammalian oocytes and embryos, but this method is not quantitative and often incompatible with live cell imaging and subsequent development. Here we have applied chemically specific, label-free coherent anti-Stokes Raman scattering (CARS) microscopy to mouse oocytes and pre-implantation embryos. We show that CARS imaging can quantify the size, number and spatial distribution of lipid droplets in living mouse oocytes and embryos up to the blastocyst stage. Notably, it can be used in a way that does not compromise oocyte maturation or embryo development. We have also correlated CARS with two-photon fluorescence microscopy simultaneously acquired using fluorescent lipid probes on fixed samples, and found only a partial degree of correlation, depending on the lipid probe, clearly exemplifying the limitation of lipid labelling. In addition, we show that differences in the chemical composition of lipid droplets in living oocytes matured in media supplemented with different saturated and unsaturated fatty acids can be detected using CARS hyperspectral imaging. These results demonstrate that CARS microscopy provides a novel non-invasive method of quantifying lipid content, type and spatial distribution with sub-micron resolution in living mammalian oocytes and embryos. PMID:27151947

A dysmorphology score system for assessing embryo abnormalities in rat whole embryo culture.

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Zhang, Cindy X; Danberry, Tracy; Jacobs, Mary Ann; Augustine-Rauch, Karen

2010-12-01

The rodent whole embryo culture (WEC) system is a well-established model for characterizing developmental toxicity of test compounds and conducting mechanistic studies. Laboratories have taken various approaches in describing type and severity of developmental findings of organogenesis-stage rodent embryos, but the Brown and Fabro morphological score system is commonly used as a quantitative approach. The associated score criteria is based upon developmental stage and growth parameters, where a series of embryonic structures are assessed and assigned respective scores relative to their gestational stage, with a Total Morphological Score (TMS) assigned to the embryo. This score system is beneficial because it assesses a series of stage-specific anatomical landmarks, facilitating harmonized evaluation across laboratories. Although the TMS provides a quantitative approach to assess growth and determine developmental delay, it is limited to its ability to identify and/or delineate subtle or structure-specific abnormalities. Because of this, the TMS may not be sufficiently sensitive for identifying compounds that induce structure or organ-selective effects. This study describes a distinct morphological score system called the "Dysmorphology Score System (DMS system)" that has been developed for assessing gestation day 11 (approximately 20-26 somite stage) rat embryos using numerical scores to differentiate normal from abnormal morphology and define the respective severity of dysmorphology of specific embryonic structures and organ systems. This method can also be used in scoring mouse embryos of the equivalent developmental stage. The DMS system enhances capabilities to rank-order compounds based upon teratogenic potency, conduct structure- relationships of chemicals, and develop statistical prediction models to support abbreviated developmental toxicity screens. © 2010 Wiley-Liss, Inc.

Pre-implantation implantable cardioverter defibrillator concerns and Type D personality increase the risk of mortality in patients with an implantable cardioverter defibrillator

DEFF Research Database (Denmark)

Pedersen, Susanne S.; van den Broek, Krista C; Erdman, Ruud A M

2010-01-01

Little is known about the influence of psychological factors on prognosis in implantable cardioverter defibrillator (ICD) patients. We examined the influence of the distressed personality (Type D) and pre-implantation device concerns on short-term mortality in ICD patients.......Little is known about the influence of psychological factors on prognosis in implantable cardioverter defibrillator (ICD) patients. We examined the influence of the distressed personality (Type D) and pre-implantation device concerns on short-term mortality in ICD patients....

Protective effect of Petroselinum crispum extract in abortion using prostadin-induced renal dysfunction in female rats

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Maryam Rezazad

2014-09-01

Full Text Available Objective: Present study investigated the effects of parsley extract on pregnant rat kidneys which have undergone clinical abortion using prostaglandins. The renal protective effect of parsley extract was evaluated in pregnant rats which had an abortion. Parsley was used due to its antioxidant properties. Materials and Methods:  Fifty-four female rats were divided in 9 groups of 6: control pregnant, two pregnant groups which received parsley extract and prostadin, two non-pregnant groups treated with parsley extract and prostadin, a group administered with both treatments, and three groups which received parsley extract in pre-implantation, implantation, and post-implantation periods of embryos. Ethanolic extract (5 mg/kg was given daily to animals for 18 days of pregnancy period. Parameters such as malondialdehyde (MDA, total antioxidant statues (TAS, creatinine, and urea were measured using biochemical assays. Histopathologic studies were also done with Hematoxylin-Eosin staining method. Results: After 18 days of treatment, significant differences were observed in serum creatinine, urea, and MDA and TAS levels. Kidney cross-sections showed edema in prostadin-treated rats while improvements in parsley + prostadin -treated rats were observed. Conclusion: These results suggested that ethanolic extract of Petroselinum crispum reduced the dysfunction in rats kidney caused by prostadin-induced abortion and could have beneficial effect in reducing the progression of prostaglandin-induced edema.

Studies Using an in Vitro Model Show Evidence of Involvement of Epithelial-Mesenchymal Transition of Human Endometrial Epithelial Cells in Human Embryo Implantation*

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Uchida, Hiroshi; Maruyama, Tetsuo; Nishikawa-Uchida, Sayaka; Oda, Hideyuki; Miyazaki, Kaoru; Yamasaki, Akiko; Yoshimura, Yasunori

2012-01-01

Human embryo implantation is a critical multistep process consisting of embryo apposition/adhesion, followed by penetration and invasion. Through embryo penetration, the endometrial epithelial cell barrier is disrupted and remodeled by an unknown mechanism. We have previously developed an in vitro model for human embryo implantation employing the human choriocarcinoma cell line JAR and the human endometrial adenocarcinoma cell line Ishikawa. Using this model we have shown that stimulation with ovarian steroid hormones (17β-estradiol and progesterone, E2P4) and suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, enhances the attachment and adhesion of JAR spheroids to Ishikawa. In the present study we showed that the attachment and adhesion of JAR spheroids and treatment with E2P4 or SAHA individually induce the epithelial-mesenchymal transition (EMT) in Ishikawa cells. This was evident by up-regulation of N-cadherin and vimentin, a mesenchymal cell marker, and concomitant down-regulation of E-cadherin in Ishikawa cells. Stimulation with E2P4 or SAHA accelerated Ishikawa cell motility, increased JAR spheroid outgrowth, and enhanced the unique redistribution of N-cadherin, which was most prominent in proximity to the adhered spheroids. Moreover, an N-cadherin functional blocking antibody attenuated all events but not JAR spheroid adhesion. These results collectively provide evidence suggesting that E2P4- and implanting embryo-induced EMT of endometrial epithelial cells may play a pivotal role in the subsequent processes of human embryo implantation with functional control of N-cadherin. PMID:22174415

Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring

DEFF Research Database (Denmark)

Ingerslev, Hans Jakob; Hindkjær, Johnny Juhl; Kirkegaard, Kirstine

2012-01-01

recently demonstrated to occur from first cleavage cycle in mice using time-lapse microscopy, with the largest impact on the pre-compaction stages. However, embryonic development in mice differs in many aspects from human embryonic development. The objective of this retrospective, descriptive study...... was to evaluate the influence of oxygen tension on human pre-implantation development using time-lapse monitoring. Materials and methods: Human embryos were cultured to the blastocyst stage in a time-lapse incubator (EmbryoScope™) in 20% O2 (group 1), 20% O2 for 24 hours followed by culture in 5% O2 (group 2......) or in 5% O2 (group 3). Eligible were patients with age 8 oocytes retrieved. Group 1 consisted of 120 IVF/ICSI embryos from 26 patients recruited to a study conducted to evaluate the safety of the time-lapse incubator by randomising 1:1 embryos from a patient to culture...

Pre- and Peri-/Post-Compaction Follistatin Treatment Increases In Vitro Production of Cattle Embryos.

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Guo Zhenhua

Full Text Available Our previous studies demonstrated that maternal (oocyte derived follistatin (FST expression is positively associated with bovine oocyte competence and exogenous follistatin treatment during the pre-compaction period of development (d 1-3 post insemination is stimulatory to bovine early embryogenesis in vitro [blastocyst rates and cell numbers/allocation to trophectoderm (TE]. In the present study, bovine embryos were treated with exogenous follistatin during d 1-3, d 4-7 and d 1-7 post insemination to test the hypothesis that embryotropic effects of exogenous follistatin are specific to the pre-compaction period (d 1-3 of early embryogenesis. Follistatin treatment during d 4-7 (peri-/post-compaction period of embryo culture increased proportion of embryos reaching blastocyst and expanded blastocyst stage and total cell numbers compared to controls, but blastocyst rates and total cell numbers were lower than observed following d 1-3 (pre-compaction follistatin treatment. Follistatin supplementation during d 1-7 of embryo culture increased development to blastocyst and expanded blastocyst stages and blastocyst total cell numbers compared to d 1-3 and d 4-7 follistatin treatment and untreated controls. A similar increase in blastocyst CDX2 mRNA and protein (TE cell marker was observed in response to d 1-3, d 4-7 and d 1-7 follistatin treatment. However, an elevation in blastocyst BMP4 protein (TE cell regulator was observed in response to d 1-3 and d 1-7, but not d 4-7 (peri-/post-compaction follistatin treatment. In summary, our study revealed the potential utility of follistatin treatment for increasing the success rate of in vitro embryo production in cattle. Such results also expand our understanding of the embryotropic actions of follistatin and demonstrate that follistatin actions on blastocyst development and cell allocation to the TE layer are not specific to the pre-compaction period.

Pre-persons, commodities or cyborgs: the legal construction and representation of the embryo.

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Fox, M

2000-01-01

This paper explores how embryos have been represented in law. It argues that two main models have underpinned legal discourse concerning the embryo. One discourse, which has become increasingly prevalent, views embryos as legal subjects or persons. Such representations are facilitated by technological developments such as ultrasound imaging. In addition to influencing Parliamentary debate prior to the passage of the Human Fertilisation and Embryology Act 1990, images of embryos as persons feature prominently in popular culture, including advertising and films, and this discourse came to the fore in the 'orphaned embryo' debate in 1996. The main opposing discourse dismisses embryos as commodifiable objects, which fits with a trend towards legal recognition that reproductive materials such as sperm may be classified as property which may be donated or sold. In the case of cryopreserved embryos these competing perspectives have resulted in litigation over the status of frozen embryos. In this paper I argue that it might be productive to shift the debate from this polarised dispute over whether embryos matter or not, whether they are pre-persons or commodities. Instead, I suggest that we should attempt to locate them in a biotechnological milieu, where cyborg metaphors may be utilised, and questions of how we should treat embryos would be contextualized alongside our response to other cyborgs.

Msx homeobox genes critically regulate embryo implantation by controlling paracrine signaling between uterine stroma and epithelium.

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Nallasamy, Shanmugasundaram; Li, Quanxi; Bagchi, Milan K; Bagchi, Indrani C

2012-01-01

The mammalian Msx homeobox genes, Msx1 and Msx2, encode transcription factors that control organogenesis and tissue interactions during embryonic development. We observed overlapping expression of these factors in uterine epithelial and stromal compartments of pregnant mice prior to embryo implantation. Conditional ablation of both Msx1 and Msx2 in the uterus resulted in female infertility due to a failure in implantation. In these mutant mice (Msx1/2(d/d)), the uterine epithelium exhibited persistent proliferative activity and failed to attach to the embryos. Gene expression profiling of uterine epithelium and stroma of Msx1/2(d/d) mice revealed an elevated expression of several members of the Wnt gene family in the preimplantation uterus. Increased canonical Wnt signaling in the stromal cells activated β-catenin, stimulating the production of a subset of fibroblast growth factors (FGFs) in these cells. The secreted FGFs acted in a paracrine manner via the FGF receptors in the epithelium to promote epithelial proliferation, thereby preventing differentiation of this tissue and creating a non-receptive uterus refractory to implantation. Collectively, these findings delineate a unique signaling network, involving Msx1/2, Wnts, and FGFs, which operate in the uterus at the time of implantation to control the mesenchymal-epithelial dialogue critical for successful establishment of pregnancy.

The stress response in gametes and embryos after paternal chemical exposures

International Nuclear Information System (INIS)

Hales, Barbara F.; Aguilar-Mahecha, Adriana; Robaire, Bernard

2005-01-01

There is increasing concern that paternal exposure to toxic chemicals impacts negatively on progeny outcome. Exposure of male rats to a model male-mediated developmental toxicant and anticancer alkylating agent, cyclophosphamide, resulted in increased pre- and post-implantation loss, as well as in malformations. We hypothesize that the stage specificity of the effects of paternal cyclophosphamide exposure on progeny depends on the ability of germ cells to respond to stress, repair DNA or undergo apoptosis. Acute high dose exposure of male rats to cyclophosphamide increased the expression of heat shock proteins and DNA repair genes, predominantly in round spermatids. In contrast, chronic low dose treatment dramatically decreased the expression of stress response genes in pachytene spermatocytes and round spermatids, but not in elongated spermatids; this reduced ability to respond to stress may allow damage to accumulate, resulting in altered sperm function. Increased DNA damage was maximal 3 weeks after drug exposure, during spermiogenesis, a key point in sperm chromatin remodelling. Drug exposure for 9 weeks increased the frequency of spermatozoa with chromosome 4 disomy and nullisomy. DNA damage found in cyclophosphamide-exposed spermatozoa was imparted to the newly fertilized zygote. Drug-exposed spermatozoa decondensed more rapidly than control spermatozoa and male pronuclear formation was earlier. RNA synthesis was higher in 1-cell embryos sired by drug-treated fathers than in controls. Significantly, the profile of gene expression was altered in embryos sired by drug-treated males as early as the 1-cell stage. Thus, exposure of male rats to cyclophosphamide altered male germ cell quality with a consequent temporal and spatial disruption of the zygotic genome activation

Oral Morphine Consumption Reduces Lens Development in Rat Embryos

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Hossein Bahadoran

2012-07-01

Full Text Available Objective: Consumption of morphine, during pregnancy, in addition to inducing defects in the mother’s nervous system function, caused defects or delays in the formation and evolution of embryonic visual system. In the present study, changes in lens development was assessed in embryos exposed in utero to morphine. Material and Methods: Female Wistar rats (250-300 g were mated with male rats and pregnancy was determined by sperm observation in vaginal smear. This day was considered as embryonic day zero (E0. The females were then divided randomly into the experimental and the control groups. The control group received tap water and the experimental group received morphine (0.05 mg/ml in their water. On embryonic day 13 ( E13, blood samples were collected from the retro-orbital sinus of all animals for plasma corticosterone detection. On embryonic day 17(E17, the animals were killed by an overdose of chloroform and the embryos were taken out surgically. The embryos were fixed in 10% formalin for 30 days. At this time, the head of the embryos were removed for tissue processing and Hematoxylin- Eosin (H&E staining. The samples were evaluated using light microscope and MOTIC software. Results: Our data indicated that plasma corticosterone level was dramatically increased and the lens was thinner in the experimental group. (Although the proliferation of lens cells increased in the experiment group but that lens had delay in removing the proliferated and elongation cells with abnormal density in the lateral part of the lens in compare with control group. I have no idea what the authors are stating here. Moreover, the opening of the eyelids was delayed in the off springs of the mothers who received morphine. Conclusions: This study showed that morphine consumption during pregnancy leads to defects in fetal visual system development, particularly in the lens, and eyelids.

Effects of dietary omega-3 and -6 supplementations on phospholipid fatty acid composition in mice uterus during window of pre-implantation.

Science.gov (United States)

Fattahi, Amir; Darabi, Masoud; Farzadi, Laya; Salmassi, Ali; Latifi, Zeinab; Mehdizadeh, Amir; Shaaker, Maghsood; Ghasemnejad, Tohid; Roshangar, Leila; Nouri, Mohammad

2018-03-01

Since fatty acid composition of uterus phospholipids is likely to influence embryo implantation, this study was conducted to investigate the effects of dietary omega-3 and -6 fatty acids on implantation rate as well as uterine phospholipid fatty acids composition during mice pre-implantation period. Sixty female mice were randomly distributed into:1) control (standard pellet), 2) omega-3 (standard pellet + 10% w/w of omega-3 fatty acids) and 3) omega-6 (standard pellet + 10% w/w of omega-6 fatty acids). Uterine phospholipid fatty acid composition during the pre-implantation window (days 1-5 of pregnancy) was analyzed using gas-chromatography. The implantation rate on the fifth day of pregnancy was also determined. Our results showed that on days 1, 2 and 3 of pregnancy, the levels of arachidonic acid (ARA) as well as total omega-6 fatty acids were significantly higher and the levels of linolenic acid and total omega-3 fatty acids were statistically lower in the omega-6 group compared to the omega-3 group (p omega-6 fatty acids, and poly-unsaturated fatty acids levels were significantly different between the two dietary supplemented groups (p omega-6 fatty acids, especially ARA, with the implantation rate. The present study showed that diets rich in omega-3 and -6 fatty acids could differently modify uterine phospholipid fatty acid composition and uterine levels of phospholipid ARA, and that the total omega-6 fatty acids had a positive association with the implantation rate. Copyright © 2017 Elsevier Inc. All rights reserved.

Designer babies on tap? Medical students' attitudes to pre-implantation genetic screening.

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Meisenberg, Gerhard

2009-03-01

This paper describes two studies about the determinants of attitudes to pre-implantation genetic screening in a multicultural sample of medical students from the United States. Sample sizes were 292 in study 1 and 1464 in study 2. Attitudes were of an undifferentiated nature, but respondents did make a major distinction between use for disease prevention and use for enhancement. No strong distinctions were made between embryo selection and germ line gene manipulations, and between somatic gene therapy and germ line gene manipulations. Religiosity was negatively associated with acceptance of "designer baby" technology for Christians and Muslims but not Hindus. However, the strongest and most consistent influence was an apparently moralistic stance against active and aggressive interference with natural processes in general. Trust in individuals and institutions was unrelated to acceptance of the technology, indicating that fear of abuse by irresponsible individuals and corporations is not an important determinant of opposition.

Uterine inactivation of muscle segment homeobox (Msx) genes alters epithelial cell junction proteins during embryo implantation

OpenAIRE

Sun, Xiaofei; Park, Craig B.; Deng, Wenbo; Potter, S. Steven; Dey, Sudhansu K.

2015-01-01

Embryo implantation requires that the uterus differentiate into the receptive state. Failure to attain uterine receptivity will impede blastocyst attachment and result in a compromised pregnancy. The molecular mechanism by which the uterus transitions from the prereceptive to the receptive stage is complex, involving an intricate interplay of various molecules. We recently found that mice with uterine deletion of Msx genes (Msx1d/d/Msx2d/d) are infertile because of implantation failure associ...

Amorphous clusters in Co implanted ZnO induced by boron pre-implantation

Energy Technology Data Exchange (ETDEWEB)

Potzger, K.; Shalimov, A.; Zhou, S.; Schmidt, H.; Mucklich, A.; Helm, M.; Fassbender, J.; Liberati, M.; Arenholz, E.

2009-02-09

We demonstrate the formation of superparamagnetic/ferromagnetic regions within ZnO(0001) single crystals sequently implanted with B and Co. While the pre-implantation with B plays a minor role for the electrical transport properties, its presence leads to the formation of amorphous phases. Moreover, B acts strongly reducing on the implanted Co. Thus, the origin of the ferromagnetic ordering in local clusters with large Co concentration is itinerant d-electrons as in the case of metallic Co. The metallic amorphous phases are non-detectable by common X-ray diffraction.

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

Science.gov (United States)

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

2016-01-01

STUDY QUESTION Can a generally applicable morphokinetic algorithm suitable for Day 3 transfers of time-lapse monitored embryos originating from different culture conditions and fertilization methods be developed for the purpose of supporting the embryologist's decision on which embryo to transfer back to the patient in assisted reproduction? SUMMARY ANSWER The algorithm presented here can be used independently of culture conditions and fertilization method and provides predictive power not surpassed by other published algorithms for ranking embryos according to their blastocyst formation potential. WHAT IS KNOWN ALREADY Generally applicable algorithms have so far been developed only for predicting blastocyst formation. A number of clinics have reported validated implantation prediction algorithms, which have been developed based on clinic-specific culture conditions and clinical environment. However, a generally applicable embryo evaluation algorithm based on actual implantation outcome has not yet been reported. STUDY DESIGN, SIZE, DURATION Retrospective evaluation of data extracted from a database of known implantation data (KID) originating from 3275 embryos transferred on Day 3 conducted in 24 clinics between 2009 and 2014. The data represented different culture conditions (reduced and ambient oxygen with various culture medium strategies) and fertilization methods (IVF, ICSI). The capability to predict blastocyst formation was evaluated on an independent set of morphokinetic data from 11 218 embryos which had been cultured to Day 5. PARTICIPANTS/MATERIALS, SETTING, METHODS The algorithm was developed by applying automated recursive partitioning to a large number of annotation types and derived equations, progressing to a five-fold cross-validation test of the complete data set and a validation test of different incubation conditions and fertilization methods. The results were expressed as receiver operating characteristics curves using the area under the

A microcontroller-based implantable nerve stimulator used for rats.

Science.gov (United States)

Sha, Hong; Zheng, Zheng; Wang, Yan; Ren, Chaoshi

2005-01-01

A microcontroller-based stimulator that can be flexible programmed after it has been implanted into a rat was studied. Programmability enables implanted stimulators to generate customized, complex protocols for experiments. After implantation, a coded light pulse train that contains information of specific identification will unlock a certain stimulator. If a command that changing the parameters is received, the microcontroller will update its flash memory after it affirms the commands. The whole size of it is only 1.6 cubic centimeters, and it can work for a month. The devices have been successfully used in animal behavior experiments, especially on rats.

Msx homeobox genes critically regulate embryo implantation by controlling paracrine signaling between uterine stroma and epithelium.

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Shanmugasundaram Nallasamy

Full Text Available The mammalian Msx homeobox genes, Msx1 and Msx2, encode transcription factors that control organogenesis and tissue interactions during embryonic development. We observed overlapping expression of these factors in uterine epithelial and stromal compartments of pregnant mice prior to embryo implantation. Conditional ablation of both Msx1 and Msx2 in the uterus resulted in female infertility due to a failure in implantation. In these mutant mice (Msx1/2(d/d, the uterine epithelium exhibited persistent proliferative activity and failed to attach to the embryos. Gene expression profiling of uterine epithelium and stroma of Msx1/2(d/d mice revealed an elevated expression of several members of the Wnt gene family in the preimplantation uterus. Increased canonical Wnt signaling in the stromal cells activated β-catenin, stimulating the production of a subset of fibroblast growth factors (FGFs in these cells. The secreted FGFs acted in a paracrine manner via the FGF receptors in the epithelium to promote epithelial proliferation, thereby preventing differentiation of this tissue and creating a non-receptive uterus refractory to implantation. Collectively, these findings delineate a unique signaling network, involving Msx1/2, Wnts, and FGFs, which operate in the uterus at the time of implantation to control the mesenchymal-epithelial dialogue critical for successful establishment of pregnancy.

Desenvolvimento pré-embrionário em ratas tratadas com oxcarbazepina nos quatro primeiros dias após a inseminação Pre-embryo development in rats treated with oxcarbazepine in the first four days after insemination

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M. de O. Guerra

2000-10-01

Full Text Available A oxcarbazepina é uma droga antiepiléptica de alta eficácia e poucos efeitos colaterais, mas pouco estudada quanto a seus efeitos durante a gestação humana e animal. OBJETIVO: Verificar se a administração de oxcarbazepine em ratas, nos quatro primeiros dias após a inseminação, altera a viabilidade ou o desenvolvimento do pré-embrião. MÉTODOS: Ratas Wistar foram tratadas com 20 ou 200 mg de oxcarbazepina/ Kg de peso corporal, via gástrica, nos dias 1, 2, 3, ou 4 a partir da inseminação ou, consecutivamente, do 1º ao 4º. Os pré-embriões foram coletados no quinto dia, visando verificar a quantidade e o desenvolvimento até a fase de blastocisto expandido. O peso corporal materno e sinais como pelos eriçados e alteração de atividade locomotora foram anotados para verificar indícios de toxicidade materna. Número de corpos lúteos e peso de ovários foram anotados com vistas à capacidade reprodutiva do animal. RESULTADOS: Não ocorreram perdas de pesos corporais maternos e nenhuma alteração física indicativa de desconforto para as ratas. Peso de ovários e número de corpos lúteos não diferiram entre tratados e controles. O número médio de pré-embriões por mães, o índice de perdas embrionárias, a proporção de blastocistos expandidos com relação ao total de pré-embriões e a média de blastocistos expandidos/mãe, não diferiram entre tratados e controles. CONCLUSÃO: A oxcarbazepina administrada em ratas, seguindo o esquema terapêutico mencionado, não apresentou efeito tóxico sobre a mãe e não alterou o desenvolvimento do pré-embrião.Oxcarbazepine is a highly efficcacious antiepileptic drug which has very few side effects and has been poorly investigated as to its effects during human and animal gestation. PURPOSE: To verify if the administration of oxcapazepine to female rats in the first four days ofter fertilization alters the viability or development of the pre-embryo. METHODS: Wistar rats were

The Effects of Progesterone on Oocyte Maturation and Embryo Development

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Saeed Zavareh

2013-01-01

Full Text Available Oocyte maturation and embryo development are controlled by intra-ovarian factors suchas steroid hormones. Progesterone (P4 exists in the follicular fluid that contributes tonormal mammalian ovarian function and has several critical functions during embryodevelopment and implantation, including endometrial receptivity, embryonic survivalduring gestation and transformation of the endometrial stromal cells to decidual cells.It is well known that the physiological effects of P4 during the pre-implantation stages ofsome mammal’s embryos are mediated by P4 receptors and their gene expression is determined.The effects of P4 on oocytes and embryo development have been assessed bysome investigations, with contradictory results. P4, a dominant steroid in follicular fluidat approximately 18 hours after the luteinizing hormone (LH surge may have a criticalrole in maturation of oocytes at the germinal stage. However, it has been shown that differentconcentrations of P4 could not improve in vitro maturation rates of germinal vesicles(GV in cumulus oocyte complexes (COCs and cumulus denuded oocytes (CDOs.Culture media supplemented with P4 significantly improved mouse embryo development.In addition, an in vivo experimental design has shown high blastocyst survival andimplantation rates in P4-treated mice.In this review we explain some of the findings that pertain to the effects of P4 onoocyte maturation and embryo development both in vitro and in vivo.

Control of the heart rate of rat embryos during the organogenic period

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Ritchie HE

2016-11-01

Full Text Available Helen E Ritchie,1 Carolina Ragnerstam,2 Elin Gustafsson,2 Johanna M Jonsson,2 William S Webster2 1Discipline of Biomedical Science, Sydney Medical School, University of Sydney, Lidcombe, 2Department of Anatomy and Histology, Sydney Medical School, University of Sydney, Sydney, NSW, Australia Abstract: The aim of this study was to gain insight into whether the first trimester embryo could control its own heart rate (HR in response to hypoxia. The gestational day 13 rat embryo is a good model for the human embryo at 5–6 weeks gestation, as the heart is comparable in development and, like the human embryo, has no functional autonomic nerve supply at this stage. Utilizing a whole-embryo culture technique, we examined the effects of different pharmacological agents on HR under normoxic (95% oxygen and hypoxic (20% oxygen conditions. Oxygen concentrations ≤60% caused a concentration-dependent decrease in HR from normal levels of ~210 bpm. An adenosine agonist, AMP-activated protein kinase (AMPK activator and KATP channel opener all caused bradycardia in normoxic conditions; however, putative antagonists for these systems failed to prevent or ameliorate hypoxia-induced bradycardia. This suggests that the activation of one or more of these systems is not the primary cause of the observed hypoxia-induced bradycardia. Inhibition of oxidative phosphorylation also decreased HR in normoxic conditions, highlighting the importance of ATP levels. The β-blocker metoprolol caused a concentration-dependent reduction in HR supporting reports that β1-adrenergic receptors are present in the early rat embryonic heart. The cAMP inducer colforsin induced a positive chronotropic effect in both normoxic and hypoxic conditions. Overall, the embryonic HR at this stage of development is responsive to the level of oxygenation, probably as a consequence of its influence on ATP production. Keywords: embryonic heart rate, embryo, bradycardia, in vitro, ATP, hypoxia

Influence of radiation (Co60) in pre-implant rabbit embryos: effect on mitotic index and embryonic pole malformations

International Nuclear Information System (INIS)

Approbato, M.S.; Moura, K.K.V.O.; Florencio, R.S.; Cunha Junior, C.; Garcia, R.; Faria, R.S.; Benedetti, L.N.; Goulart, F.B.

1995-01-01

We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), at three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses: five c Gy and ten c Gy. Six rabbits (36 blastocysts) were used as controls. the matching instant was the zero hour. Exactly six days after (± 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. the embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryo parameters were studied: mitotic index; embryonic pole malformations. There were no gross abnormalities of embryo pole. The mitotic index were altered both by the time and doses. (author)

Melatonin promotes the in vitro development of pronuclear embryos and increases the efficiency of blastocyst implantation in murine.

Science.gov (United States)

Wang, Feng; Tian, XiuZhi; Zhang, Lu; Tan, DunXian; Reiter, Russel J; Liu, GuoShi

2013-10-01

When a defect occurs in the in vitro development of a pronuclear embryo, the interruption of the subsequent implantation limits the success of assisted conception. This common problem remains to be solved. In this study, we observed that melatonin at its physiological concentration (10(-7)  m) significantly promoted the in vitro development of murine pronuclear embryos. This was indicated by the increased blastocyst rate, hatching blastocyst rate, and blastocyst cell number with melatonin treatment. In addition, when these blastocysts were implanted into female recipient mice, the pregnancy rates (95.0% versus control 67.8%), litter sizes (4.1 pups/litter versus control 2.7 pups/litter), and postnatal survival rates of offspring (96.84% versus control 81.24%) were significantly improved compared with their non-melatonin-treated counterparts. Mechanistic studies revealed that melatonin treatment upregulates gene expression of the antioxidant enzyme, superoxide dismutase (SOD), and the anti-apoptotic factor bcl-2 while downregulating the expression of pro-apoptotic genes p53 and caspase-3. Due to these changes, melatonin treatment reduces ROS production and cellular apoptosis during in vitro embryo development and improves the quality of blastocysts. The implantation of blastocysts with higher quality leads to more healthy offspring and increased pup survival. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Potential hazards to embryo implantation: A human endometrial in vitro model to identify unwanted antigestagenic actions of chemicals

International Nuclear Information System (INIS)

Fischer, L.; Deppert, W.R.; Pfeifer, D.; Stanzel, S.; Weimer, M.; Hanjalic-Beck, A.; Stein, A.; Straßer, M.; Zahradnik, H.P.; Schaefer, W.R.

2012-01-01

Embryo implantation is a crucial step in human reproduction and depends on the timely development of a receptive endometrium. The human endometrium is unique among adult tissues due to its dynamic alterations during each menstrual cycle. It hosts the implantation process which is governed by progesterone, whereas 17β-estradiol regulates the preceding proliferation of the endometrium. The receptors for both steroids are targets for drugs and endocrine disrupting chemicals. Chemicals with unwanted antigestagenic actions are potentially hazardous to embryo implantation since many pharmaceutical antiprogestins adversely affect endometrial receptivity. This risk can be addressed by human tissue-specific in vitro assays. As working basis we compiled data on chemicals interacting with the PR. In our experimental work, we developed a flexible in vitro model based on human endometrial Ishikawa cells. Effects of antiprogestin compounds on pre-selected target genes were characterized by sigmoidal concentration–response curves obtained by RT-qPCR. The estrogen sulfotransferase (SULT1E1) was identified as the most responsive target gene by microarray analysis. The agonistic effect of progesterone on SULT1E1 mRNA was concentration-dependently antagonized by RU486 (mifepristone) and ZK137316 and, with lower potency, by 4-nonylphenol, bisphenol A and apigenin. The negative control methyl acetoacetate showed no effect. The effects of progesterone and RU486 were confirmed on the protein level by Western blotting. We demonstrated proof of principle that our Ishikawa model is suitable to study quantitatively effects of antiprogestin-like chemicals on endometrial target genes in comparison to pharmaceutical reference compounds. This test is useful for hazard identification and may contribute to reduce animal studies. -- Highlights: ► We compare progesterone receptor-mediated endometrial effects of chemicals and drugs. ► 4-Nonylphenol, bisphenol A and apigenin exert weak

Potential hazards to embryo implantation: A human endometrial in vitro model to identify unwanted antigestagenic actions of chemicals

Energy Technology Data Exchange (ETDEWEB)

Fischer, L.; Deppert, W.R. [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany); Pfeifer, D. [Department of Hematology and Oncology, University Hospital Freiburg (Germany); Stanzel, S.; Weimer, M. [Department of Biostatistics, German Cancer Research Center, Heidelberg (Germany); Hanjalic-Beck, A.; Stein, A.; Straßer, M.; Zahradnik, H.P. [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany); Schaefer, W.R., E-mail: wolfgang.schaefer@uniklinik-freiburg.de [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany)

2012-05-01

Embryo implantation is a crucial step in human reproduction and depends on the timely development of a receptive endometrium. The human endometrium is unique among adult tissues due to its dynamic alterations during each menstrual cycle. It hosts the implantation process which is governed by progesterone, whereas 17β-estradiol regulates the preceding proliferation of the endometrium. The receptors for both steroids are targets for drugs and endocrine disrupting chemicals. Chemicals with unwanted antigestagenic actions are potentially hazardous to embryo implantation since many pharmaceutical antiprogestins adversely affect endometrial receptivity. This risk can be addressed by human tissue-specific in vitro assays. As working basis we compiled data on chemicals interacting with the PR. In our experimental work, we developed a flexible in vitro model based on human endometrial Ishikawa cells. Effects of antiprogestin compounds on pre-selected target genes were characterized by sigmoidal concentration–response curves obtained by RT-qPCR. The estrogen sulfotransferase (SULT1E1) was identified as the most responsive target gene by microarray analysis. The agonistic effect of progesterone on SULT1E1 mRNA was concentration-dependently antagonized by RU486 (mifepristone) and ZK137316 and, with lower potency, by 4-nonylphenol, bisphenol A and apigenin. The negative control methyl acetoacetate showed no effect. The effects of progesterone and RU486 were confirmed on the protein level by Western blotting. We demonstrated proof of principle that our Ishikawa model is suitable to study quantitatively effects of antiprogestin-like chemicals on endometrial target genes in comparison to pharmaceutical reference compounds. This test is useful for hazard identification and may contribute to reduce animal studies. -- Highlights: ► We compare progesterone receptor-mediated endometrial effects of chemicals and drugs. ► 4-Nonylphenol, bisphenol A and apigenin exert weak

No cytotoxic effects from application of pentoxifylline to spermatozoa on subsequent pre-implantation embryo development in mice

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Mohammad Ali Khalili

2017-06-01

Full Text Available The aim was to assess the effect of spermatozoa exposed to PTX on the rates of fertilization and embryo development and apoptotic cells within blastocysts in an animal model. Mice Oocytes were inseminated with spermatozoa exposed to 3.6 mmol PTX for 30 min, or with neat spermatozoa. Then fertilization and embryo development rate, blastocyst formation and quality, as well as total cell number of blastocyst, and DNA fragmentation index (DFI in blastocysts were surveyed in both groups. Fertilization and embryo development rate were similar between the groups. The rates of blastocyst formation did not differ significantly between control and PTX groups (52.4% vs. 51.8%. The average of total cell count in blastocysts and DFI in control and PTX groups were also insignificant (31.08 ± 1.5 vs. 34.14 ± 1.5 and 9.76 ± 5.0 vs. 11.77 ± 5.4. Application of PTX for enhancing sperm motility does not cause a cytotoxic effect on subsequent embryo development and embryo genome integrity.

Ovarian stimulation and embryo quality

NARCIS (Netherlands)

Baart, Esther; Macklon, Nick S.; Fauser, Bart J. C. M.

To Study the effects of different ovarian stimulation approaches on oocyte and embryo quality, it is imperative to assess embryo quality with a reliable and objective method. Embryos rated as high quality by standardized morphological assessment are associated with higher implantation and pregnancy

Microphone directionality, pre-emphasis filter, and wind noise in cochlear implants.

Science.gov (United States)

Chung, King; McKibben, Nicholas

2011-10-01

Wind noise can be a nuisance or a debilitating masker for cochlear implant users in outdoor environments. Previous studies indicated that wind noise at the microphone/hearing aid output had high levels of low-frequency energy and the amount of noise generated is related to the microphone directionality. Currently, cochlear implants only offer either directional microphones or omnidirectional microphones for users at-large. As all cochlear implants utilize pre-emphasis filters to reduce low-frequency energy before the signal is encoded, effective wind noise reduction algorithms for hearing aids might not be applicable for cochlear implants. The purposes of this study were to investigate the effect of microphone directionality on speech recognition and perceived sound quality of cochlear implant users in wind noise and to derive effective wind noise reduction strategies for cochlear implants. A repeated-measure design was used to examine the effects of spectral and temporal masking created by wind noise recorded through directional and omnidirectional microphones and the effects of pre-emphasis filters on cochlear implant performance. A digital hearing aid was programmed to have linear amplification and relatively flat in-situ frequency responses for the directional and omnidirectional modes. The hearing aid output was then recorded from 0 to 360° at flow velocities of 4.5 and 13.5 m/sec in a quiet wind tunnel. Sixteen postlingually deafened adult cochlear implant listeners who reported to be able to communicate on the phone with friends and family without text messages participated in the study. Cochlear implant users listened to speech in wind noise recorded at locations that the directional and omnidirectional microphones yielded the lowest noise levels. Cochlear implant listeners repeated the sentences and rated the sound quality of the testing materials. Spectral and temporal characteristics of flow noise, as well as speech and/or noise characteristics before

Uterine inactivation of muscle segment homeobox (Msx) genes alters epithelial cell junction proteins during embryo implantation.

Science.gov (United States)

Sun, Xiaofei; Park, Craig B; Deng, Wenbo; Potter, S Steven; Dey, Sudhansu K

2016-04-01

Embryo implantation requires that the uterus differentiate into the receptive state. Failure to attain uterine receptivity will impede blastocyst attachment and result in a compromised pregnancy. The molecular mechanism by which the uterus transitions from the prereceptive to the receptive stage is complex, involving an intricate interplay of various molecules. We recently found that mice with uterine deletion ofMsxgenes (Msx1(d/d)/Msx2(d/d)) are infertile because of implantation failure associated with heightened apicobasal polarity of luminal epithelial cells during the receptive period. However, information on Msx's roles in regulating epithelial polarity remains limited. To gain further insight, we analyzed cell-type-specific gene expression by RNA sequencing of separated luminal epithelial and stromal cells by laser capture microdissection fromMsx1(d/d)/Msx2(d/d)and floxed mouse uteri on d 4 of pseudopregnancy. We found that claudin-1, a tight junction protein, and small proline-rich (Sprr2) protein, a major component of cornified envelopes in keratinized epidermis, were substantially up-regulated inMsx1(d/d)/Msx2(d/d)uterine epithelia. These factors also exhibited unique epithelial expression patterns at the implantation chamber (crypt) inMsx1(f/f)/Msx2(f/f)females; the patterns were lost inMsx1(d/d)/Msx2(d/d)epithelia on d 5, suggesting important roles during implantation. The results suggest thatMsxgenes play important roles during uterine receptivity including modulation of epithelial junctional activity.-Sun, X., Park, C. B., Deng, W., Potter, S. S., Dey, S. K. Uterine inactivation of muscle segment homeobox (Msx) genes alters epithelial cell junction proteins during embryo implantation. © FASEB.

The Effect of a GnRH Agonist Injection or Progesterone Implant at Diestrus in Cryopreserved Embryo Transferred Cows

OpenAIRE

KIRBAŞ, Mesut; BÜLBÜL, Bülent; KÖSE, Mehmet; DURSUN, Şükrü; ÇOLAK, Mehmet

2014-01-01

In this study, the effect of a single dose of GnRH on d 13 or progesterone implant for 7 days between d 13 and 20 on plasma progesterone levels and pregnancy rates on cryopreserved embryo transferred cows were investigated. Synchronized 48 Brown Swiss recipient cows were used as animal material. Seven days after estrus detection, cryopreserved cattle embryos were transferred into recipients and cows were assigned randomly into three groups. In GnRH group (n=16), cows were intramuscularly inje...

In vitro and in vivo Development of Cloned Ovine Embryos using in vitro and in vivo Matured Oocytes

DEFF Research Database (Denmark)

Holm, P; Nagashima, H; Sun, F-J

1995-01-01

Cloning of sheep embryos by nucleus transplantation can be achieved by using in vivo matured (oviductal) oocytes and in vivo culture. However, these steps involve cumbersome procedures. Therefore, the effects of in vivo vs. the equivalent in vitro procedures on the pre-implantation development...... matured oocytes were enucleated and fused with inserted blastomeres from donor embryos. In vitro matured oocytes were enucleated and allowed to age prior to blastomere insertion and electrofusion. Fused embryos were cultured for approximately 132 h either in vivo in ligated sheep oviducts or in vitro...

Possible sources of neuroprotection following subretinal silicon chip implantation in RCS rats

Science.gov (United States)

Pardue, Machelle T.; Phillips, Michael J.; Yin, Hang; Fernandes, Alcides; Cheng, Yian; Chow, Alan Y.; Ball, Sherry L.

2005-03-01

Current retinal prosthetics are designed to stimulate existing neural circuits in diseased retinas to create a visual signal. However, implantation of retinal prosthetics may create a neurotrophic environment that also leads to improvements in visual function. Possible sources of increased neuroprotective effects on the retina may arise from electrical activity generated by the prosthetic, mechanical injury due to surgical implantation, and/or presence of a chronic foreign body. This study evaluates these three neuroprotective sources by implanting Royal College of Surgeons (RCS) rats, a model of retinitis pigmentosa, with a subretinal implant at an early stage of photoreceptor degeneration. Treatment groups included rats implanted with active and inactive devices, as well as sham-operated. These groups were compared to unoperated controls. Evaluation of retinal function throughout an 18 week post-implantation period demonstrated transient functional improvements in eyes implanted with an inactive device at 6, 12 and 14 weeks post-implantation. However, the number of photoreceptors located directly over or around the implant or sham incision was significantly increased in eyes implanted with an active or inactive device or sham-operated. These results indicate that in the RCS rat localized neuroprotection of photoreceptors from mechanical injury or a chronic foreign body may provide similar results to subretinal electrical stimulation at the current output evaluated here.

Improved Bone Micro Architecture Healing Time after Implant Surgery in an Ovariectomized Rat.

Science.gov (United States)

Takahashi, Takahiro; Watanabe, Takehiro; Nakada, Hiroshi; Sato, Hiroki; Tanimoto, Yasuhiro; Sakae, Toshiro; Kimoto, Suguru; Mijares, Dindo; Zhang, Yu; Kawai, Yasuhiko

2016-01-01

The present animal study investigated whether oral intake of synthetic bone mineral (SBM) improves peri-implant bone formation and bone micro architecture (BMA). SBM was used as an intervention experimental diet and AIN-93M was used as a control. The SBM was prepared by mixing dicalcium phosphate dihydrate (CaHPO 4 ·2H 2 O) and magnesium and zinc chlorides (MgCl 2 and ZnCl 2 , respectively), and hydrolyzed in double-distilled water containing dissolved potassium carbonate and sodium fluoride. All rats were randomly allocated into one of two groups: a control group was fed without SBM (n = 18) or an experimental group was fed with SBM (n = 18), at seven weeks old. At 9 weeks old, all rats underwent implant surgery on their femurs under general anesthesia. The implant was inserted into the insertion socket prepared at rats' femur to a depth of 2.5 mm by using a drill at 500 rpm. Nine rats in each group were randomly selected and euthanized at 2 weeks after implantation. The remaining nine rats in each group continued their diets, and were euthanized in the same manner at 4 weeks after implantation. The femur, including the implant, was removed from the body and implant was pulled out by an Instron universal testing machine. After the implant removal, BMA was evaluated by bone surface ratio (BS/BV), bone volume fraction (BV/TV), trabecular thickness (TbTh), trabecular number (TbN), trabecular star volume (Vtr), and micro-CT images. BS/BV, BV/TV, TbTh and Vtr were significantly greater in the rats were fed with SBM than those were fed without SBM at 2 and 4 weeks after implantation (P implant formation and BMA, prominent with trabecular bone structure. The effect of SBM to improve secondary stability of the implant, and shortening the treatment period should be investigated in the future study.

The Effects of Maternal Hyperthyroidism on Histologic Changes in Parietal Lobe in Rat Embryos

OpenAIRE

Fatemeh Mirsafi; Gholamreza Kaka; Mahnaz Azarnia

2017-01-01

Background Maternal hyperthyroidism causes developmental defects on the nervous system of fetuses. Objectives The present study was designed to study the effects of maternal hyperthyroidism on the development of the parietal lobe in the brain of rat embryos. Methods In this experimental study, thirty Sprague-Dawley rats were randomly divided into three groups. The control group rec...

Determination of metallic traces in kidneys, livers, lungs and spleens of rats with metallic implants after a long implantation time.

Science.gov (United States)

Rubio, Juan Carlos; Garcia-Alonso, Maria Cristina; Alonso, Concepcion; Alobera, Miguel Angel; Clemente, Celia; Munuera, Luis; Escudero, Maria Lorenza

2008-01-01

Metallic transfer from implants does not stop at surrounding tissues, and metallic elements may be transferred by proteins to become lodged in organs far from the implant. This work presents an in vivo study of metallic implant corrosion to measure metallic element accumulation in organs located far from the implant, such as kidneys, livers, lungs and spleens. The studied metallic implant materials were CoCr alloy, Ti, and the experimental alloy MA956 coated with alpha-alumina. The implants were inserted in the hind legs of Wistar rats. Analysis for Co, Cr, Ti and Al metallic traces was performed after a long exposure time of 12 months by Inductively Coupled Plasma (ICP) with Mass Spectrometry (MS). According to the results, the highest Cr and Ti concentrations were detected in spleens. Co is mainly found in kidneys, since this element is eliminated via urine. Cr and Ti traces increased significantly in rat organs after the long implantation time. The organs of rats implanted with the alpha-alumina coated experimental MA956 did not present any variation in Al content after 12 months, which means there was no degradation of the alumina layer surface.

RepSox improves viability and regulates gene expression in rhesus monkey-pig interspecies cloned embryos.

Science.gov (United States)

Zhu, Hai-Ying; Jin, Long; Guo, Qing; Luo, Zhao-Bo; Li, Xiao-Chen; Zhang, Yu-Chen; Xing, Xiao-Xu; Xuan, Mei-Fu; Zhang, Guang-Lei; Luo, Qi-Rong; Wang, Jun-Xia; Cui, Cheng-Du; Li, Wen-Xue; Cui, Zheng-Yun; Yin, Xi-Jun; Kang, Jin-Dan

2017-05-01

To investigate the effect of the small molecule, RepSox, on the expression of developmentally important genes and the pre-implantation development of rhesus monkey-pig interspecies somatic cell nuclear transfer (iSCNT) embryos. Rhesus monkey cells expressing the monomeric red fluorescent protein 1 which have a normal (42) chromosome complement, were used as donor cells to generate iSCNT embryos. RepSox increased the expression levels of the pluripotency-related genes, Oct4 and Nanog (p  0.05), this was not significant. RepSox can improve the developmental potential of rhesus monkey-pig iSCNT embryos by regulating the expression of pluripotency-related genes.

In vitro and in vivo Development of Cloned Ovine Embryos using in vitro and in vivo Matured Oocytes

DEFF Research Database (Denmark)

Holm, P; Nagashima, H; Sun, F-J

1995-01-01

Cloning of sheep embryos by nucleus transplantation can be achieved by using in vivo matured (oviductal) oocytes and in vivo culture. However, these steps involve cumbersome procedures. Therefore, the effects of in vivo vs. the equivalent in vitro procedures on the pre-implantation development of...

[Pre-operation evaluation and intra-operation management of cochlear implantation].

Science.gov (United States)

Zhang, Dao-xing; Hu, Bao-hua; Xiao, Yu-li; Shi, Bo-ning

2004-10-01

To summarize pre-operation evaluation experiences in cochlear implantation. Performing auditory evaluation and image analysis seriously in 158 severe hearing loss or total deaf cases before cochlear implantation, comparing their performance with the findings during and post operation. Among the total 158 cases, 116 cases with normal structure, 42 cases with the abnormal findings of the inner or middle ear. Stapedial gusher happened in 6 cases, 1 case was not predicted before operation. Except 1 case with serious malformation, the findings of other 157 cases in operation were consistent with the pre-operation evaluation. We helped all patients reconstruct auditory conduction with cochlear implantation, and the average hearing level up to 37.6 dB SPL. Performing image analysis seriously before operation and planning for operation according to HRCT can do great help to cochlear implantation. The operation under the HRCT instruction has less complications.

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

DEFF Research Database (Denmark)

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

2005-01-01

The processes of cellular differentiation were studied in somatic cell nuvlear transfer (SCNT), in vitro cultured (IVC) and in vivo developed (in vivo) ovine embryos on days 7, 9, 11, 13, 17 and 19. SCNT embryos were constructed from in vitro matured oocytes and granulosa cells, and IVC embryos...... were produced by in vitro culture of in vivo fertilized zygotes. Most SCNT and IVC embryos were transferred to recipients on day 6 while some remained in culture for day 7 processing. In vivo embryos were collected as zygotes, transferred to intermediate recipients and retransferred to final recipients...

Neutron irradiation of rat embryos in utero

International Nuclear Information System (INIS)

Vogel, H.H. Jr.

1978-01-01

In the rat radiation is most effective in producing congenital anomalies during the organ-forming period (days 9 to 13), which is approximately equivalent to the 14th to 50th days of human pregnancy. We have exposed female Sprague--Dawley rats on the 18th day of pregnancy to single whole-body doses of fission neutrons (20 to 150 rads). After 20 rads there was a small decrease in body weight which lasted from birth to weaning. During this period 9% of the irradiated rats died compared with 4% of the controls. After 50 rads, 65/275 (23.6%) of the rats died between birth and weaning, and the body-weight loss of the survivors was increased. After 100 rads, 62/133 (47%) died at birth or day 1 and 103/133 (77.4%) died before weaning. A large and significant decrease in body weight persisted in the survivors. After 150 rads of fission neutrons, all 95 rats died within 48 hr of birth. From cross-fostering experiments, we believe this is a direct effect of radiation on the embryos and not an indirect action through the mother or her milk. The LD 50 for the period from birth to weaning is approximately 75 rads of fission neutrons. Studies of organ weight were conducted daily for the first week after birth in an attempt to find the cause of radiation mortality. Body weight of the irradiated animals averaged only about one-half that of the controls. The liver, kidney, brain, and testes of the neutron-irradiated rats weighed significantly less than those of the controls. The weights of the spleen, lungs, duodenum, and stomach were decreased but not significantly. The bone marrow appeared depleted in the irradiated long bones, but the spleen maintained active hematopoiesis 1 to 2 months after neutron exposure

Association of the transcription profile of bovine oocytes and embryos with developmental potential

Czech Academy of Sciences Publication Activity Database

Kaňka, Jiří; Němcová, Lucie; Toralová, Tereza; Vodičková Kepková, Kateřina; Vodička, Petr; Jeseta, M.; Machatková, M.

2012-01-01

Roč. 134, 1-2 (2012), 29-35 ISSN 0378-4320. [Embryo Genomics Meeting /3./. Bonn, 20.08.2012-22.08.2012] R&D Projects: GA ČR GA523/09/1035; GA MZe QI91A018 Institutional support: RVO:67985904 Keywords : oocyte * in vitro maturation * pre-implantation development Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.897, year: 2012

Comparative effects of sodium channel blockers in short term rat whole embryo culture

Energy Technology Data Exchange (ETDEWEB)

Nilsson, Mats F, E-mail: Mats.Nilsson@farmbio.uu.se [Department of Pharmaceutical Biosciences, Uppsala University (Sweden); Sköld, Anna-Carin; Ericson, Ann-Christin; Annas, Anita; Villar, Rodrigo Palma [AstraZeneca R and D Södertälje (Sweden); Cebers, Gvido [AstraZeneca R and D, iMed, 141 Portland Street, Cambridge, MA 02139 (United States); Hellmold, Heike; Gustafson, Anne-Lee [AstraZeneca R and D Södertälje (Sweden); Webster, William S [Department of Anatomy and Histology, University of Sydney (Australia)

2013-10-15

This study was undertaken to examine the effect on the rat embryonic heart of two experimental drugs (AZA and AZB) which are known to block the sodium channel Nav1.5, the hERG potassium channel and the L-type calcium channel. The sodium channel blockers bupivacaine, lidocaine, and the L-type calcium channel blocker nifedipine were used as reference substances. The experimental model was the gestational day (GD) 13 rat embryo cultured in vitro. In this model the embryonic heart activity can be directly observed, recorded and analyzed using computer assisted image analysis as it responds to the addition of test drugs. The effect on the heart was studied for a range of concentrations and for a duration up to 3 h. The results showed that AZA and AZB caused a concentration-dependent bradycardia of the embryonic heart and at high concentrations heart block. These effects were reversible on washout. In terms of potency to cause bradycardia the compounds were ranked AZB > bupivacaine > AZA > lidocaine > nifedipine. Comparison with results from previous studies with more specific ion channel blockers suggests that the primary effect of AZA and AZB was sodium channel blockage. The study shows that the short-term rat whole embryo culture (WEC) is a suitable system to detect substances hazardous to the embryonic heart. - Highlights: • Study of the effect of sodium channel blocking drugs on embryonic heart function • We used a modified method rat whole embryo culture with image analysis. • The drugs tested caused a concentration dependent bradycardia and heart block. • The effect of drugs acting on multiple ion channels is difficult to predict. • This method may be used to detect cardiotoxicity in prenatal development.

Comparative effects of sodium channel blockers in short term rat whole embryo culture

International Nuclear Information System (INIS)

Nilsson, Mats F; Sköld, Anna-Carin; Ericson, Ann-Christin; Annas, Anita; Villar, Rodrigo Palma; Cebers, Gvido; Hellmold, Heike; Gustafson, Anne-Lee; Webster, William S

2013-01-01

This study was undertaken to examine the effect on the rat embryonic heart of two experimental drugs (AZA and AZB) which are known to block the sodium channel Nav1.5, the hERG potassium channel and the L-type calcium channel. The sodium channel blockers bupivacaine, lidocaine, and the L-type calcium channel blocker nifedipine were used as reference substances. The experimental model was the gestational day (GD) 13 rat embryo cultured in vitro. In this model the embryonic heart activity can be directly observed, recorded and analyzed using computer assisted image analysis as it responds to the addition of test drugs. The effect on the heart was studied for a range of concentrations and for a duration up to 3 h. The results showed that AZA and AZB caused a concentration-dependent bradycardia of the embryonic heart and at high concentrations heart block. These effects were reversible on washout. In terms of potency to cause bradycardia the compounds were ranked AZB > bupivacaine > AZA > lidocaine > nifedipine. Comparison with results from previous studies with more specific ion channel blockers suggests that the primary effect of AZA and AZB was sodium channel blockage. The study shows that the short-term rat whole embryo culture (WEC) is a suitable system to detect substances hazardous to the embryonic heart. - Highlights: • Study of the effect of sodium channel blocking drugs on embryonic heart function • We used a modified method rat whole embryo culture with image analysis. • The drugs tested caused a concentration dependent bradycardia and heart block. • The effect of drugs acting on multiple ion channels is difficult to predict. • This method may be used to detect cardiotoxicity in prenatal development

Embryo-fetal development toxicity of honokiol microemulsion intravenously administered to pregnant rats.

Science.gov (United States)

Zhang, Qianqian; Ye, Xiangfeng; Wang, Lingzhi; Peng, Bangjie; Zhang, Yingxue; Bao, Jie; Li, Wanfang; Wei, Jinfeng; Wang, Aiping; Jin, Hongtao; Chen, Shizhong

2016-02-01

The aim of this study was to evaluate the embryo-fetal development toxicity of honokiol microemulsion. The drug was intravenously injected to pregnant SD rats at dose levels of 0, 200, 600 and 2000 μg/kg/day from day 6-15 of gestation. All the pregnant animals were observed for body weights and any abnormal changes and subjected to caesarean-section on gestation day (GD) 20; all fetuses obtained from caesarean-section were assessed by external inspection, visceral and skeletal examinations. No treatment-related external alterations as well as visceral and skeletal malformations were observed in honokiol microemulsion groups. There was no significant difference in the body weight gain of the pregnant rats, average number of corpora lutea, and the gravid uterus weight in the honokiol microemulsion groups compared with the vehicle control group. However, at a dose level of 2000 μg/kg/day, there was embryo-fetal developmental toxicity observed, including a decrease in the body length and tail length of fetuses. In conclusion, the no-observed-adverse-effect level (NOAEL) of honokiol microemulsion is 600 μg/kg/day, 75 times above the therapeutic dosage and it has embryo-fetal toxicity at a dose level of 2000 μg/kg/day, which is approximately 250 times above the therapeutic dosage. Copyright © 2015 Elsevier Inc. All rights reserved.

Hypothesis: Co-transfer of genuine embryos and implantation-promoting compounds via artificial containers improve endometrium receptivity.

Science.gov (United States)

Celik, Onder; Acet, Mustafa; Celik, Sudenaz; Sahin, Levent; Koc, Onder; Celik, Nilufer

2017-06-01

As with other organs endometrial functions are altered with the advancing age. Age related decrease in reproductive functions leads to decline in the number of oocytes retrieved and the synthesis of endometrial receptivity molecules. Despite the significant improvement in assisted reproductive technologies we do not have so many options to enhance endometrial receptivity. Due to lack of drugs having endometrium receptivity enhancement properties, oocyte donation seems to be the only solution for women with implantation failure. The euploid oocytes come from young and healthy donors may overcome age associated endometrial receptivity defect. Nevertheless, many reasons restrict us from using oocyte donation in women with implantation failure. We, therefore, hypothesized that by mimicking a young blastocyst's effect on endometrium, the transfer of genuine embryos and implantation-promoting compounds together might be the new treatment option for infertile women with recurrent implantation failure. Artificial beads, MI or GV oocytes, and empty zona can be used as a container for intrauterine replacement of implantation-promoting compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Effect of low dose aspirin on osseointegration around titanium implants in osteoporotic rats].

Science.gov (United States)

Yang, Q; Li, F L

2018-02-09

Objective: To investigate the effect of aspirin on osseointegration around titanium implants in ostoeporotic rats and to provide evidence for future researches and clinical application. Methods: A total of 60 female SD rats, aged 3-4 months, were divided into ovariectomy group (Ovx group, n= 48) and sham-ovariectomy group (Sham group, n= 12). The rats in Ovx group received ovariectomy and those in Sham group underwent sham-ovariectomy. Twelve weeks later, six rats in each group were randomly selected to confirm the osteoporosis models. The Ovx group was divided into 4 subgroups with 12 rats in each group, namely the osteoporosis group (OP group), and Aspirin groups (A1, A2, A3 group). Pure screw titanium implants were placed in the right tibia near metaphysis of all rats. Three days after implant surgery, aspirin groups were intragastrically administered aspirin at a dose of 2.06, 4.11, 8.21 mg·kg(-1)·d(-1) (A1, A2, A3), and OP group and Sham group were fed the same amount of normal saline. Four and 12 weeks following implantations surgery, half of the rats in each group were randomly chosen and sacrificed. Implant bone contact rate (IBCR), combined bone lamella width (CBLW) and trabercular width (TW) were observed and calculated using histomorphometric measurement. Results: Four weeks after implantations surgery, the TW and CBLW of rats in A1 group [(39.60±2.77) and (27.56±4.14) μm] and the IBCR, TW and CBLW of rats in A2 group and A3 group [A2: (47.21±4.19)%, (48.74±3.20) and (35.91±3.79) μm; A3: (47.35±6.07)%, (50.27±5.25) and (40.66±2.11) μm] were much higher than those in OP group [(33.89±7.17)%, (32.20±6.10) and (19.77±6.80) μm]( P 0.05). Twelve weeks after implantations surgery, the IBCR and CBLW of rats in A1 group [ (85.86±3.64) %, (53.12±8.68) μm], and the IBCR, TW and CBLW of rats in A2 group and A3 group [A2: (85.64±3.97)%, (69.42±6.78) and (54.19±3.12) μm; A3: (86.22±3.48)%, (75.43±3.50) and (55.79±5.60) μm] were much higher

Supplementation of fetal bovine serum alters histone modification H3R26me2 during preimplantation development of in vitro produced bovine embryos

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Daniel R. Arnold

2015-07-01

Full Text Available Abstract In vitro production (IVP of bovine embryos is not only of great economic importance to the cattle industry, but is also an important model for studying embryo development. The aim of this study was to evaluate the histone modification, H3R26me2 during pre-implantation development of IVP bovine embryos cultured with or without serum supplementation and how these in vitro treatments compared to in vivo embryos at the morula stage. After in vitro maturation and fertilization, bovine embryos were cultured with either 0 or 2.5% fetal bovine serum (FBS. Development was evaluated and embryos were collected and fixed at different stages during development (2-, 4-, 8-, 16-cell, morula and blastocyst. Fixed embryos were then used for immunofluorescence utilizing an antibody for H3R26me2. Images of stained embryos were analyzed as a percentage of total DNA. Embryos cultured with 2.5% FBS developed to blastocysts at a greater rate than 0%FBS groups (34.85±5.43% vs. 23.38±2.93%; P<0.05. Levels of H3R26me2 changed for both groups over development. In the 0%FBS group, the greatest amount of H3R26me2 staining was at the 4-cell (P<0.05, 16-cell (P<0.05 and morula (P<0.05 stages. In the 2.5%FBS group, only 4-cell stage embryos were significantly higher than all other stages (P<0.01. Morula stage in vivo embryos had similar levels as the 0%FBS group, and both were significantly higher than the 2.5%FBS group. These results suggest that the histone modification H3R26me2 is regulated during development of pre-implantation bovine embryos, and that culture conditions greatly alter this regulation.

Effect of pre-implanted oxygen in Si on the retention of implanted He

Energy Technology Data Exchange (ETDEWEB)

Manuaba, A. [KFKI Research Institute for Particle and Nuclear Physics, P.O. Box 49, H-1525 Budapest (Hungary); Paszti, F. [KFKI Research Institute for Particle and Nuclear Physics, P.O. Box 49, H-1525 Budapest (Hungary)]. E-mail: paszti@rmki.kfki.hu; Ramos, A.R. [ITN - Instituto Tecnologico e Nuclear, Estrada Nacional 10, P-2686-953, Sacavem (Portugal); Khanh, N.Q. [MTA Research Institute for Technical Physics and Materials Science, P.O. Box 49, H-1525 Budapest (Hungary); Pecz, B. [MTA Research Institute for Technical Physics and Materials Science, P.O. Box 49, H-1525 Budapest (Hungary); Zolnai, Z. [MTA Research Institute for Technical Physics and Materials Science, P.O. Box 49, H-1525 Budapest (Hungary); Tunyogi, A. [KFKI Research Institute for Particle and Nuclear Physics, P.O. Box 49, H-1525 Budapest (Hungary); Szilagyi, E. [KFKI Research Institute for Particle and Nuclear Physics, P.O. Box 49, H-1525 Budapest (Hungary)

2006-08-15

Buried SiO {sub x} layers, with different x values, were formed by implanting 80 keV O{sup +} ions with different fluences into single crystal Si samples at room temperature. Into each of these O pre-implanted layers, 20 keV He{sup +} was implanted up to the fluence of 1 x 10{sup 17} ion/cm{sup 2}. The He distribution profiles were determined by 2045 keV proton backscattering spectrometry. It was found that as the O content increases, the retained He gradually decreases at the beginning, then rapidly falls at x = 0.6 till it disappears at x = 1.3. The process that leads to this phenomenon is discussed.

Does gestrinone antagonize the effects of estrogen on endometrial implants upon the peritoneum of rats?

Directory of Open Access Journals (Sweden)

Vera Lúcia Rodrigues Lobo

2008-01-01

Full Text Available OBJECTIVE: To evaluate the effects of estrogen treatment in combination with gestrinone on an experimental rat model of endometriosis. METHODS: Uterine transplants were attached to the peritoneum of female Wistar rats via a surgical autotransplantation technique. The implanted area was measured during the proestrus phase and after hormonal treatment. We performed morphometric analysis and examined the macroscopic and morphometric alterations of endometrial implants after hormonal treatment in ovariectomized rats. RESULTS: The high dose of estrogen caused macroscopic increases in the endometrial implant group compared with other groups, which were similar to increases in the proestrus phase. The low dose showed morphometric development of implants, such as an increase in number of endometrial glands, leukocyte infiltration and mitosis. Gestrinone antagonized both doses of estrogen. CONCLUSION: Our findings suggest that gestrinone antagonizes estrogen's effects on rat peritoneal endometrial implants.

Ethanol impedes embryo transport and impairs oviduct epithelium

International Nuclear Information System (INIS)

Xu, Tonghui; Yang, Qiuhong; Liu, Ruoxi; Wang, Wenfu; Wang, Shuanglian; Liu, Chuanyong; Li, Jingxin

2016-01-01

Most studies have demonstrated that alcohol consumption is associated with decreased fertility. The aim of this study was to investigate the effects of alcohol on pre-implantation embryo transport and/or early embryo development in the oviduct. We reported here that ethanol concentration-dependently suppressed the spontaneous motility of isolated human oviduct strips (EC50 50 ± 6 mM), which was largely attenuated in the present of L-NAME, a classical nitric oxide synthase(NOS) competitive inhibitor. Notably, either acute or chronic alcohol intake delayed egg transport and retarded early development of the embryo in the mouse oviduct, which was largely rescued by co-administration of L-NAME in a acute alcohol intake group but not in chronic alcohol intake group. It is worth mentioning that the oviductal epithelium destruction was verified by scanning electron microscope (SEM) observations in chronic alcohol intake group. In conclusion, alcohol intake delayed egg transport and retarded early development of the embryo in the oviduct by suppressing the spontaneous motility of oviduct and/or impairing oviductal epithelium. These findings suggested that alcohol abuse increases the incident of ectopic pregnancy.

Ethanol impedes embryo transport and impairs oviduct epithelium.

Science.gov (United States)

Xu, Tonghui; Yang, Qiuhong; Liu, Ruoxi; Wang, Wenfu; Wang, Shuanglian; Liu, Chuanyong; Li, Jingxin

2016-05-16

Most studies have demonstrated that alcohol consumption is associated with decreased fertility. The aim of this study was to investigate the effects of alcohol on pre-implantation embryo transport and/or early embryo development in the oviduct. We reported here that ethanol concentration-dependently suppressed the spontaneous motility of isolated human oviduct strips (EC50 50±6mM), which was largely attenuated in the present of L-NAME, a classical nitric oxide synthase(NOS) competitive inhibitor. Notably, either acute or chronic alcohol intake delayed egg transport and retarded early development of the embryo in the mouse oviduct, which was largely rescued by co-administration of L-NAME in a acute alcohol intake group but not in chronic alcohol intake group. It is worth mentioning that the oviductal epithelium destruction was verified by scanning electron microscope (SEM) observations in chronic alcohol intake group. In conclusion, alcohol intake delayed egg transport and retarded early development of the embryo in the oviduct by suppressing the spontaneous motility of oviduct and/or impairing oviductal epithelium. These findings suggested that alcohol abuse increases the incident of ectopic pregnancy. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Time Course of Peri-Implant Bone Regeneration around Loaded and Unloaded Implants in a Rat Model

Science.gov (United States)

Jariwala, Shailly H.; Wee, Hwabok; Roush, Evan P.; Whitcomb, Tiffany L.; Murter, Christopher; Kozlansky, Gery; Lakhtakia, Akhlesh; Kunselman, Allen R.; Donahue, Henry J.; Armstrong, April D.; Lewis, Gregory S.

2018-01-01

The time-course of cancellous bone regeneration surrounding mechanically loaded implants affects implant fixation, and is relevant to determining optimal rehabilitation protocols following orthopaedic surgeries. We investigated the influence of controlled mechanical loading of titanium-coated polyether-ether ketone (PEEK) implants on osseointegration using time-lapsed, non-invasive, in vivo micro-computed tomography (micro-CT) scans. Implants were inserted into proximal tibial metaphyses of both limbs of eight female Sprague-Dawley rats. External cyclic loading (60 μm or 100 μm displacement, 1 Hz, 60 seconds) was applied every other day for 14 days to one implant in each rat, while implants in contralateral limbs served as the unloaded controls. Hind limbs were imaged with high-resolution micro-CT (12.5 μm voxel size) at 2, 5, 9, and 12 days post-surgery. Trabecular changes over time were detected by 3D image registration allowing for measurements of bone-formation rate (BFR) and bone-resorption rate (BRR). At day 9, mean %BV/TV for loaded and unloaded limbs were 35.5 ± 10.0 % and 37.2 ± 10.0 %, respectively, and demonstrated significant increases in bone volume compared to day 2. BRR increased significantly after day 9. No significant differences between bone volumes, BFR, and BRR were detected due to implant loading. Although not reaching significance (p = 0.16), an average 119 % increase in pull-out strength was measured in the loaded implants. PMID:27381807

Evaluation of porous vitreous carbon or silicon implants by radiology in rat's skull

International Nuclear Information System (INIS)

Vaccari-Mazzetti, Marcelo Paulo; Kobata, Celio Toshiro; Fabiani, Paulo; Martins, Dulce Maria Fonseca Soares; Gomes, Paulo de Oliveira; Martins, Jose Luiz

2008-01-01

Purpose: Evaluate by CT the use of porous vitreous carbon (PVC) and silicon (S) implants as the replacement bone in the craniofacial skeleton of rats. Methods: 40 rats divided in: Group A (n=20) PVC submitted to the implant of a fragment in skull. After the euthanasia, the animals were divided into two subgroups: A I: 10 animals, studied in the 7th postoperative day (P.O) and AII: 10 animals, studied in the 28th P.O. In group B, S, 20 rats were submitted to S implant in the skull. All other steps were identical to group A, with designation of subgroups BI and BII. CT with beams in axial cuts of 1 mm thickness to obtain 3-D information It was used Hounsfield scale for evaluate the radio density of the implant. They were used non parametric tests to analyze the results. Results: The 7th PO boss remained in the two groups, but for 28th PO, observed reduction in the volume of the implant in Group A, not observed in group B. CT studies noticed different radio densities around all of S prostheses (pseudo capsule), that do not appeared in CPV implants. The S has remained unchanged in the CT, but the CPV has had a modification in its radio density (p≤0,05), in all implants. Conclusion: In CT evaluation the implants of CPV have greater deformation that the S, which makes them not suitable for replacement of membranous bone in the rat skull. (author)

Behavior of the P1.HTR mastocytoma cell line implanted in the chorioallantoic membrane of chick embryos

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S.F. Avram

2013-01-01

Full Text Available The P1.HTR cell line includes highly transfectable cells derived from P815 mastocytoma cells originating from mouse breast tissue. Despite its widespread use in immunogenic studies, no data are available about the behavior of P1.HTR cells in the chick embryo chorioallantoic membrane model. The objective of the present investigation was to study the effects of P1.HTR cells implanted on the chorioallantoic membrane of chick embryos. We inoculated P1.HTR cells into the previously prepared chick embryo chorioallantoic membrane and observed the early and late effects of these cells by stereomicroscopy, histochemistry and immunohistochemistry. A highly angiotropic and angiogenic effect occurred early after inoculation and a tumorigenic potential with the development of mastocytoma keeping well mast cells immunophenotype was detected later during the development. The P1.HTR mastocytoma cell line is a good tool for the development of the chick embryo chorioallantoic membrane mastocytoma model and also for other studies concerning the involvement of blood vessels. The chick embryo chorioallantoic membrane model of mastocytoma retains the mast cell immunophenotype under experimental conditions and could be used as an experimental tool for in vivo preliminary testing of antitumor and antivascular drugs.

Accumulation of oocytes and/or embryos by vitrification: a new strategy for managing poor responder patients undergoing pre implantation diagnosis [v2; ref status: indexed, http://f1000r.es/321

Directory of Open Access Journals (Sweden)

Alexia Chatziparasidou

2014-03-01

Full Text Available Background: Low (or poor responder patients are women who require large doses of stimulation medications and produce less than an optimal number of oocytes during IVF cycles. Low responder patients produce few oocytes and embryos, which significantly reduces their chances for success in a preimplantation genetic diagnosis (PGD cycle. Accumulation of vitrified oocytes or embryos before the actual PGD cycle is a possible strategy that might increase patient’s chances for a healthy pregnancy. Aim of the study: This retrospective study evaluates the efficacy of a PGD program in low responder patients after repeated ovarian stimulation cycles with cumulative vitrification of oocytes and embryos. Methods: Over a period of 30 months, 13 patients entering the PGD program were identified as poor responders after their first ovarian stimulation. These patients started a PGD cycle for one of the following indications: history of recurrent implantation failure (n=1, cystic fibrosis (n=1, X-linked microtubular myopathy (n=1, recurrent miscarriages (n=5, Duchene muscular dystrophy (n=1, chromosomal translocation (n=1 and high sperm aneuploidy (n=1.  After multiple ovarian hormonal stimulations patients had either all mature oocytes (Group A; 3 patients or all of their day 2 embryos vitrified (group B; 10 patients. Mean total number of oocyte collections per patient was 2.3 (range: 2 - 5 cycles. Results: In the actual PGD cycle, all vitrified oocytes from group A patients were warmed and underwent intra cytoplasmic sperm injection (ICSI followed by culture up to day 3. For group B patients all vitrified day 2 embryos were warmed and cultured overnight. On day 3 of culture, all embryos from Group A and B had blastomere biopsy followed by genetic analysis. In group A, 20 embryos were found suitable for biopsy and genetic analysis; at least one healthy embryo was available for transfer for each patient.  For group B, 72 embryos in total were available for

Where does New Zealand stand on permitting research on human embryos?

Science.gov (United States)

Jones, D Gareth

2014-08-01

In many respects New Zealand has responded to the assisted reproductive technologies (ARTs) as positively as many comparable societies, such as Australia and the UK. Consequently, in vitro fertilisation (IVF) and pre-implantation genetic diagnosis (PGD) are widely available, as is non-commercial surrogacy utilising IVF. These developments have been made possible by the Human Assisted Reproductive Technology (HART) Act 2004, overseen by its two committees, the Advisory Committee on Assisted Reproductive Technology (ACART) and the Ethics Committee (ECART). However, New Zealand stands apart from many of these other societies by the lack of permission for scientists to conduct research using human embryos. There is no doubt this reflects strongly held viewpoints on the part of some that embryos should be protected and not exploited. Legitimate as this stance is, the resulting situation is problematic when IVF is already designated as an established procedure. This is because the development of IVF involved embryo research, and continuing improvements in procedures depend upon ongoing embryo research. While prohibition of research on human embryos gives the impression of protecting embryos, it fails to do this and also fails to enhance the health and wellbeing of children born using IVF. This situation will not be rectified until research is allowed on human embryos.

SU-E-I-42: Normalized Embryo/fetus Doses for Fluoroscopically Guided Pacemaker Implantation Procedures Calculated Using a Monte Carlo Technique

Energy Technology Data Exchange (ETDEWEB)

Damilakis, J; Stratakis, J; Solomou, G [University of Crete, Heraklion (Greece)

2014-06-01

Purpose: It is well known that pacemaker implantation is sometimes needed in pregnant patients with symptomatic bradycardia. To our knowledge, there is no reported experience regarding radiation doses to the unborn child resulting from fluoroscopy during pacemaker implantation. The purpose of the current study was to develop a method for estimating embryo/fetus dose from fluoroscopically guided pacemaker implantation procedures performed on pregnant patients during all trimesters of gestation. Methods: The Monte Carlo N-Particle (MCNP) radiation transport code was employed in this study. Three mathematical anthropomorphic phantoms representing the average pregnant patient at the first, second and third trimesters of gestation were generated using Bodybuilder software (White Rock science, White Rock, NM). The normalized embryo/fetus dose from the posteroanterior (PA), the 30° left-anterior oblique (LAO) and the 30° right-anterior oblique (RAO) projections were calculated for a wide range of kVp (50–120 kVp) and total filtration values (2.5–9.0 mm Al). Results: The results consist of radiation doses normalized to a) entrance skin dose (ESD) and b) dose area product (DAP) so that the dose to the unborn child from any fluoroscopic technique and x-ray device used can be calculated. ESD normalized doses ranged from 0.008 (PA, first trimester) to 2.519 μGy/mGy (RAO, third trimester). DAP normalized doses ranged from 0.051 (PA, first trimester) to 12.852 μGy/Gycm2 (RAO, third trimester). Conclusion: Embryo/fetus doses from fluoroscopically guided pacemaker implantation procedures performed on pregnant patients during all stages of gestation can be estimated using the method developed in this study. This study was supported by the Greek Ministry of Education and Religious Affairs, General Secretariat for Research and Technology, Operational Program ‘Education and Lifelong Learning’, ARISTIA (Research project: CONCERT)

SU-E-I-42: Normalized Embryo/fetus Doses for Fluoroscopically Guided Pacemaker Implantation Procedures Calculated Using a Monte Carlo Technique

International Nuclear Information System (INIS)

Damilakis, J; Stratakis, J; Solomou, G

2014-01-01

Purpose: It is well known that pacemaker implantation is sometimes needed in pregnant patients with symptomatic bradycardia. To our knowledge, there is no reported experience regarding radiation doses to the unborn child resulting from fluoroscopy during pacemaker implantation. The purpose of the current study was to develop a method for estimating embryo/fetus dose from fluoroscopically guided pacemaker implantation procedures performed on pregnant patients during all trimesters of gestation. Methods: The Monte Carlo N-Particle (MCNP) radiation transport code was employed in this study. Three mathematical anthropomorphic phantoms representing the average pregnant patient at the first, second and third trimesters of gestation were generated using Bodybuilder software (White Rock science, White Rock, NM). The normalized embryo/fetus dose from the posteroanterior (PA), the 30° left-anterior oblique (LAO) and the 30° right-anterior oblique (RAO) projections were calculated for a wide range of kVp (50–120 kVp) and total filtration values (2.5–9.0 mm Al). Results: The results consist of radiation doses normalized to a) entrance skin dose (ESD) and b) dose area product (DAP) so that the dose to the unborn child from any fluoroscopic technique and x-ray device used can be calculated. ESD normalized doses ranged from 0.008 (PA, first trimester) to 2.519 μGy/mGy (RAO, third trimester). DAP normalized doses ranged from 0.051 (PA, first trimester) to 12.852 μGy/Gycm2 (RAO, third trimester). Conclusion: Embryo/fetus doses from fluoroscopically guided pacemaker implantation procedures performed on pregnant patients during all stages of gestation can be estimated using the method developed in this study. This study was supported by the Greek Ministry of Education and Religious Affairs, General Secretariat for Research and Technology, Operational Program ‘Education and Lifelong Learning’, ARISTIA (Research project: CONCERT)

Significance of buccopalatal implant position, biotype, platform switching, and pre-implant bone augmentation on the level of the midbuccal mucosa

NARCIS (Netherlands)

Zuiderveld, Elise G; den Hartog, Laurens; Vissink, Arjan; Raghoebar, Gerry M; Meijer, Henny J A

2014-01-01

This study assessed whether buccopalatal implant position, biotype, platform switching, and pre-implant bone augmentation affects the level of the midbuccal mucosa (MBM). Ninety patients with a single-tooth implant in the esthetic zone were included. The level of the MBM was measured on photographs

Action of uranium on pre implanted mouse embryos; Accion del uranio sobre los embriones de preimplantacion de raton

Energy Technology Data Exchange (ETDEWEB)

Kundt, Miriam S [Comision Nacional de Energia Atomica, General San Martin (Argentina). Dept. de Radiobiologia

2001-07-01

The cultured preimplantation embryos are normally employed to evaluate the effects of environmental pollutants specially metals. Embryos were obtained from hybrid females CBA x C57 Bl following induction of super ovulation. They were incubated from 1 cell stage during 120 hs. in M16 cultured medium. Three different experiments were carried out: A, B and C using uranyl nitrate UO{sub 2}(NO{sub 3}){sub 2} 6H{sub 2}O as source of uranium. In experiment 'A' the embryos were cultivated in the same culture dish containing final U concentrations of 13, 26, 52, 104 and 208 {mu}gU/ml. In experiment 'B' embryos in a one cell stage were placed in culture medium with uranyl nitrate with final U concentrations of 26, 52, 104 {mu}gU/ml. After 24 hours those embryos which had reached the two-cell stage were transferred to another culture dish to which fresh solutions of uranyl nitrate were added, maintaining the same concentrations of the previous one. In experiment 'C' the embryos were cultivated containing final U concentrations of 26, 52 and 104 {mu}gU/ml and they were transferred to another culture dish every day to which fresh solutions of uranyl nitrate were added. Different embryos parameters were analyzed: 1) Development grade; 2) Number of cell per embryo and metaphases index; and 3) Embryo ploidy. 1) Embryos were observed each 24 hs. to evaluate development grade: 2, 4 and 8 cell stage, morula, early -expanded- hatched blastocysts and atresic embryos. No significant differences were observed in the proportion of embryos arrested either in the one-cell or in the two cell stages in control culture medium regarding different concentrations of U, in a total of 4388 embryos analyzed. From 2 cell stage, moment that the embryo begins to synthesize its own ARNm, the delay in embryonic development increased dose dependent. On the other hand, the toxicological effects in the same concentration are increase from 'A' treatment to 'C' treatment. Embriotoxicology effects are

Effects of pore size, implantation time and nano-surface properties on rat skin ingrowth into percutaneous porous titanium implants

OpenAIRE

Farrell, Brad J.; Prilutsky, Boris I.; Ritter, Jana M.; Kelley, Sean; Popat, Ketul; Pitkin, Mark

2013-01-01

The main problem of percutaneous osseointegrated implants is poor skin-implant integration, which may cause infection. This study investigated the effects of pore size (Small, 40–100 microns and Large, 100–160 microns), nanotubular surface treatment (Nano), and duration of implantation (3 and 6 weeks) on skin ingrowth into porous titanium. Each implant type was percutaneously inserted in the back of 35 rats randomly assigned to 7 groups. Implant extrusion rate was measured w...

Preparation and biocompatibility study of in situ forming polymer implants in rat brains.

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Nasongkla, Norased; Boongird, Atthaporn; Hongeng, Suradej; Manaspon, Chawan; Larbcharoensub, Noppadol

2012-02-01

We describe the development of polymer implants that were designed to solidify once injected into rat brains. These implants comprised of glycofurol and copolymers of D: ,L: -lactide (LA), ε-caprolactone and poly(ethylene glycol) (PLECs). Scanning electron microscopy (SEM) and gel permeation chromatography (GPC) showed that the extent of implant degradation was increased with LA: content in copolymers. SEM analysis revealed the formation of porosity on implant surface as the degradation proceeds. PLEC with 19.3% mole of LA: was chosen to inject in rat brains at the volume of 10, 25 and 40 μl. Body weights, hematological and histopathological data of rats treated with implants were evaluated on day 3, 6, 14, 30 and 45 after the injection. Polymer solution at the injection volume of 10 μl were tolerated relatively well compared to those of 25 and 40 μl as confirmed by higher body weight and healing action (fibrosis tissue) 30 days after treatment. The results from this study suggest a possible application as drug delivery systems that can bypass the blood brain barrier.

Micro-morphologic changes around biophysically-stimulated titanium implants in ovariectomized rats

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Chang Ting-Ling

2007-07-01

Full Text Available Abstract Background Osteoporosis may present a risk factor in achievement of osseointegration because of its impact on bone remodeling properties of skeletal phsiology. The purpose of this study was to evaluate micro-morphological changes in bone around titanium implants exposed to mechanical and electrical-energy in osteoporotic rats. Methods Fifteen 12-week old sprague-dowley rats were ovariectomized to develop osteoporosis. After 8 weeks of healing period, two titanium implants were bilaterally placed in the proximal metaphyses of tibia. The animals were randomly divided into a control group and biophysically-stimulated two test groups with five animals in each group. In the first test group, a pulsed electromagnetic field (PEMF stimulation was administrated at a 0.2 mT 4 h/day, whereas the second group received low-magnitude high-frequency mechanical vibration (MECHVIB at 50 Hz 14 min/day. Following completion of two week treatment period, all animals were sacrificed. Bone sites including implants were sectioned, removed en bloc and analyzed using a microCT unit. Relative bone volume and bone micro-structural parameters were evaluated for 144 μm wide peri-implant volume of interest (VOI. Results Mean relative bone volume in the peri-implant VOI around implants PEMF and MECHVIB was significantly higher than of those in control (P P > .05 while the difference in trabecular-number among test and control groups was significant in all VOIs (P Conclusion Biophysical stimulation remarkably enhances bone volume around titanium implants placed in osteoporotic rats. Low-magnitude high-frequency MECHVIB is more effective than PEMF on bone healing in terms of relative bone volume.

Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring

DEFF Research Database (Denmark)

Ingerslev, Hans Jakob; Hindkjær, Johnny Juhl; Kirkegaard, Kirstine

2012-01-01

-points for each cell division and blastocyst stages were registered until 120 hours after oocyte retrieval. Only 2PN embryos completing the first cleavage were evaluated. The groups were compared using one-way ANOVA or Kruskall-Wallis test. Estimates are reported as medians with 95% confidence intervals. Time......Introduction: Data from a number of studies indicate -but not unequivocally- that culture of embryos in 5% O2 compared to 20% O2 improves blastocyst formation in humans and various animal species and may yield better pregnancy rates in IVF. The detrimental effects of atmospheric oxygen were...... was to evaluate the influence of oxygen tension on human pre-implantation development using time-lapse monitoring. Materials and methods: Human embryos were cultured to the blastocyst stage in a time-lapse incubator (EmbryoScope™) in 20% O2 (group 1), 20% O2 for 24 hours followed by culture in 5% O2 (group 2...

Functional studies of signaling pathways in peri-implantation development of the mouse embryo by RNAi

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Bell Graham

2005-12-01

Full Text Available Abstract Background Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post-implantation stages. Results We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post-implantation development and defects in the anterior midline and neural folds at headfold stages. Conclusion Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo.

Soluble CD146, an innovative and non-invasive biomarker of embryo selection for in vitro fertilization.

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Sylvie Bouvier

Full Text Available Although progress was made in in vitro fertilization (IVF techniques, the majority of embryos transferred fail to implant. Morphology embryo scoring is the standard procedure for most of IVF centres for choosing the best embryo, but remains limited since even the embryos classified as "top quality" may not implant. As it has been shown that i CD146 is involved in embryo implantation and ii membrane form is shed to generate soluble CD146 (sCD146, we propose that sCD146 in embryo supernatants may constitute a new biomarker of embryo selection. Immunocytochemical staining showed expression of CD146 in early embryo stages and sCD146 was detected by ELISA and Western-blot in embryo supernatants from D2. We retrospectively studied 126 couples who underwent IVF attempt. The embryo culture medium from each transferred embryo (n = 222 was collected for measurement of sCD146 by ELISA. Significantly higher sCD146 concentrations were present in embryo supernatants that did not implant (n = 185 as compared to those that successfully implanted (n = 37 (1310 +/- 1152 pg.mL-1 vs. 845+/- 1173 pg.mL-1, p = 0.024. Sensitivity analysis performed on single embryo transfers (n = 71 confirmed this association (p = 0.0054. The computed ROC curve established that the optimal sCD146 concentration for embryo implantation is under 1164 pg.mL-1 (sensitivity: 76%, specificity: 48%, PPV: 25% and NPV: 92%. Over this sCD146 threshold, the implantation rate was significantly lower (9% with sCD146 levels >1164 pg.ml-1 vs. 22% with sCD146 levels ≤ 1164 pg.mL-1, p = 0.01. Among the embryos preselected by morphologic scoring, sCD146 determination could allow a better selection of the embryo(s, thus improving the success of elective single embryo transfer. This study establishes the proof of concept for the use of sCD146 as a biomarker for IVF by excluding the embryo with the highest sCD146 level. A multicentre prospective study will now be necessary to further establish its use in

Establishment of reproducible osteosarcoma rat model using orthotopic implantation technique.

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Yu, Zhe; Sun, Honghui; Fan, Qingyu; Long, Hua; Yang, Tongtao; Ma, Bao'an

2009-05-01

In experimental musculoskeletal oncology, there remains a need for animal models that can be used to assess the efficacy of new and innovative treatment methodologies for bone tumors. Rat plays a very important role in the bone field especially in the evaluation of metabolic bone diseases. The objective of this study was to develop a rat osteosarcoma model for evaluation of new surgical and molecular methods of treatment for extremity sarcoma. One hundred male SD rats weighing 125.45+/-8.19 g were divided into 5 groups and anesthetized intraperitoneally with 10% chloral hydrate. Orthotopic implantation models of rat osteosarcoma were performed by injecting directly into the SD rat femur with a needle for inoculation with SD tumor cells. In the first step of the experiment, 2x10(5) to 1x10(6) UMR106 cells in 50 microl were injected intraosseously into median or distal part of the femoral shaft and the tumor take rate was determined. The second stage consisted of determining tumor volume, correlating findings from ultrasound with findings from necropsia and determining time of survival. In the third stage, the orthotopically implanted tumors and lung nodules were resected entirely, sectioned, and then counter stained with hematoxylin and eosin for histopathologic evaluation. The tumor take rate was 100% for implants with 8x10(5) tumor cells or more, which was much less than the amount required for subcutaneous implantation, with a high lung metastasis rate of 93.0%. Ultrasound and necropsia findings matched closely (r=0.942; p<0.01), which demonstrated that Doppler ultrasonography is a convenient and reliable technique for measuring cancer at any stage. Tumor growth curve showed that orthotopically implanted tumors expanded vigorously with time-lapse, especially in the first 3 weeks. The median time of survival was 38 days and surgical mortality was 0%. The UMR106 cell line has strong carcinogenic capability and high lung metastasis frequency. The present rat

Embryo selection: the role of time-lapse monitoring.

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Kovacs, Peter

2014-12-15

In vitro fertilization has been available for over 3 decades. Its use is becoming more widespread worldwide, and in the developed world, up to 5% of children have been born following IVF. It is estimated that over 5 million children have been conceived in vitro. In addition to giving hope to infertile couples to have their own family, in vitro fertilization has also introduced risks as well. The risk of multiple gestation and the associated maternal and neonatal morbidity/mortality has increased significantly over the past few decades. While stricter transfer policies have eliminated the majority of the high-order multiples, these changes have not yet had much of an impact on the incidence of twins. A twin pregnancy can be avoided by the transfer of a single embryo only. However, the traditionally used method of morphologic embryo selection is not predictive enough to allow routine single embryo transfer; therefore, new screening tools are needed. Time-lapse embryo monitoring allows continuous, non-invasive embryo observation without the need to remove the embryo from optimal culturing conditions. The extra information on the cleavage pattern, morphologic changes and embryo development dynamics could help us identify embryos with a higher implantation potential. These technologic improvements enable us to objectively select the embryo(s) for transfer based on certain algorithms. In the past 5-6 years, numerous studies have been published that confirmed the safety of time-lapse technology. In addition, various markers have already been identified that are associated with the minimal likelihood of implantation and others that are predictive of blastocyst development, implantation potential, genetic health and pregnancy. Various groups have proposed different algorithms for embryo selection based on mostly retrospective data analysis. However, large prospective trials are needed to study the full benefit of these (and potentially new) algorithms before their

Effects of embryo culture media do not persist after implantation: a histological study in mice.

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Hemkemeyer, Sandra A; Schwarzer, Caroline; Boiani, Michele; Ehmcke, Jens; Le Gac, Séverine; Schlatt, Stefan; Nordhoff, Verena

2014-02-01

Is post-implantation embryonic development after blastocyst transfer affected by exposure to different assisted reproduction technology (ART) culture media? Fetal development and placental histology of ART embryos cultured in vitro in different ART media was not impaired compared with embryos grown in vivo. The application of different in vitro culture (IVC) media for human ART has an effect on birthweight of newborns. In the mouse model, differences in blastocyst formation were reported after culture in different ART media. Moreover, abnormalities in the liver and heart have been detected as a result of suboptimal IVC conditions. Fertilized oocytes from inbred and outbred breeding schemes were retrieved and either immediately transferred to foster mothers or incubated in control or human ART culture media up to the blastocyst stage prior to transfer. Placental and fetal anatomy and particularly bone development were evaluated. B6C3F1 female mice were used as oocyte donors after ovulation induction. C57Bl/6 and CD1 males were used for mating and CD1 females as foster mothers for embryo transfer. Fertilized oocytes were recovered from mated females and incubated in sequential human ART media (ISM1/ISM2 and HTF/Multiblast), in control media [KSOM(aa) and Whitten's medium] or grown in utero without IVC (zygote control). As in vivo, control B6C3F1 females were superovulated and left untreated. Fetuses and placentae were isolated by Caesarean section and analysed at 18.5 days post-coitum (dpc) for placenta composition and at 15.5 dpc for body weight, crown-rump length (CRL), fetal organ development, morphological development, total bone length and extent of bone ossification. No major differences in the number of implantation sites or in histological appearance of the placentae were detected. CRL of KSOM(aa) fetuses was higher compared with zygote control and Whitten's medium. Histological analysis of tissue sections revealed no gross morphological differences compared

Theory about the Embryo Cryo-Treatment.

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Vladimirov, Iavor K; Tacheva, Desislava; Diez, Antonio

2017-04-01

To create hypothesis, which can give a logical explanation related to the benefits of freezing/thawing embryos. Cryopreservation is not only a technology used for storing embryos, but also a method of embryo treatment that can potentially improve the success rate in infertile couples. From the analysis of multiple results in assisted reproductive technology, which have no satisfactory explanation to date, we found evidence to support a 'therapeutic' effect of the freezing/thawing of embryos on the process of recovery of the embryo and its subsequent implantation. Freezing/thawing is a way to activate the endogenous survival and repair responses in preimplantation embryos. Several molecular mechanisms can explain the higher success rate of ET using thawed embryos compared to fresh ET in women of advanced reproductive age, the higher miscarriage rate in cases of thawed blastocyst ET compared to thawed ET at early cleavage embryo, and the higher perinatal parameters of born children after thawed ET. Embryo thawing induces a stress. Controlled stress is not necessarily detrimental, because it generates a phenomenon that is counteracted by several known biological responses aimed to repair mitochondrial damage of membrane and protein misfolding. The term for favorable biological responses to low exposures to stress is called hormesis. This thesis will summarize the role of cryopreservation in the activation of a hormetic response, preserving the mitochondrial function, improving survival, and having an impact on the process of implantation, miscarriage, and the development of pregnancy.

HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

International Nuclear Information System (INIS)

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

2010-01-01

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

Estrogen-induced transcription factor EGR1 regulates c-Kit transcription in the mouse uterus to maintain uterine receptivity for embryo implantation.

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Park, Mira; Kim, Hye-Ryun; Kim, Yeon Sun; Yang, Seung Chel; Yoon, Jung Ah; Lyu, Sang Woo; Lim, Hyunjung Jade; Hong, Seok-Ho; Song, Haengseok

2018-07-15

Early growth response 1 (Egr1) is a key transcription factor that mediates the action of estrogen (E 2 ) to establish uterine receptivity for embryo implantation. However, few direct target genes of EGR1 have been identified in the uterus. Here, we demonstrated that E 2 induced EGR1-regulated transcription of c-Kit, which plays a crucial role in cell fate decisions. Spatiotemporal expression of c-Kit followed that of EGR1 in uteri of ovariectomized mice at various time points after E 2 treatment. E 2 activated ERK1/2 and p38 to induce EGR1, which then activated c-Kit expression in the uterus. EGR1 transfection produced rapid and transient induction of c-KIT in a time- and dose-dependent manner. Furthermore, luciferase assays to measure c-Kit promoter activity confirmed that a functional EGR1 binding site(s) (EBS) was located within -1 kb of the c-Kit promoter. Site-directed mutagenesis and chromatin immunoprecipitation-PCR for three putative EBS within -1 kb demonstrated that the EBS at -818/-805 was critical for EGR1-dependent c-Kit transcription. c-Kit expression was significantly increased in the uterus on day 4 and administration of Masitinib, a c-Kit inhibitor, effectively interfered with embryo implantation. Collectively, our results showed that estrogen induces transcription factor EGR1 to regulate c-Kit transcription for uterine receptivity for embryo implantation in the mouse uterus. Copyright © 2017 Elsevier B.V. All rights reserved.

Attitude towards pre-implantation genetic diagnosis for hereditary cancer

NARCIS (Netherlands)

Lammens, Chantal; Bleiker, Eveline; Aaronson, Neil; Vriends, Annette; Ausems, Margreet; Jansweijer, Maaike; Wagner, Anja; Sijmons, Rolf; van den Ouweland, Ans; van der Luijt, Rob; Spruijt, Liesbeth; Gómez García, Encarna; Ruijs, Mariëlle; Verhoef, Senno

2009-01-01

The use of pre-implantation genetic diagnosis (PGD) for hereditary cancer is subject to on-going debate, particularly among professionals. This study evaluates the attitude towards PGD and attitude-associated characteristics of those concerned: family members with a hereditary cancer predisposition.

Impact of monosodium glutamate and /or gamma irradiation on pregnant rats and their embryos

International Nuclear Information System (INIS)

Ibrahim, M.F.; Darwish, M.M.

2009-01-01

The purpose of this study is to evaluate the destructive impact of the widely used nutritional flavouring agent, monosodium glutamate (MSG) and/or radiation stress on the female rats mothers and their developing embryos as judged by the maternal biochemical and embryological morphological and histopathological lesions induced. MSG is the sodium salt of glutamic acid, widely used as a food additive and flavour enhancer in modern nutrition. MSG (4 mg/rat) was daily administered subcutaneously to pregnant female rats from the 10 th to the 15 th gestational days during which they were subjected to intermittent radiation dose levels of 0.5 Gy increments delivered every other day up to a cumulative dose of 1.5 Gy whereas investigation has been carried out one day prior to parturition. MSG and radiation dual treatment resulted in increased maternal serum levels of lipid peroxides, total lipids, triglycerides, cholesterol and sodium together with decreased calcium concentrations. consequently, the developing embryos in the uteri, due to their increased sensitivity, showed various teratological and histological impairments . MSG and/or radiation induced effects were detected as growth retardation, malformations, intrauterine death and embryonic resorption. moreover, embryonic histological examination revealed ill-shaped vertebrae with degenerated osteogenic layer together with severely degenerated neurons

Detection of SEA-type α-thalassemia in embryo biopsies by digital PCR.

Science.gov (United States)

Lee, Ta-Hsien; Hsu, Ya-Chiung; Chang, Chia Lin

2017-08-01

Accurate and efficient pre-implantation genetic diagnosis (PGD) based on the analysis of single or oligo-cells is needed for timely identification of embryos that are affected by deleterious genetic traits in in vitro fertilization (IVF) clinics. Polymerase chain reaction (PCR) is the backbone of modern genetic diagnoses, and a spectrum of PCR-based techniques have been used to detect various thalassemia mutations in prenatal diagnosis (PND) and PGD. Among thalassemias, SEA-type α-thalassemia is the most common variety found in Asia, and can lead to Bart's hydrops fetalis and serious maternal complications. To formulate an efficient digital PCR for clinical diagnosis of SEA-type α-thalassemia in cultured embryos, we conducted a pilot study to detect the α-globin and SEA-type deletion alleles in blastomere biopsies with a highly sensitive microfluidics-based digital PCR method. Genomic DNA from embryo biopsy samples were extracted, and crude DNA extracts were first amplified by a conventional PCR procedure followed by a nested PCR reaction with primers and probes that are designed for digital PCR amplification. Analysis of microfluidics-based PCR reactions showed that robust signals for normal α-globin and SEA-type deletion alleles, together with an internal control gene, can be routinely generated using crude embryo biopsies after a 10 6 -fold dilution of primary PCR products. The SEA-type deletion in cultured embryos can be sensitively diagnosed with the digital PCR procedure in clinics. The adoption of this robust PGD method could prevent the implantation of IVF embryos that are destined to develop Bart's hydrops fetalis in a timely manner. The results also help inform future development of a standard digital PCR procedure for cost-effective PGD of α-thalassemia in a standard IVF clinic. Copyright © 2017. Published by Elsevier B.V.

Circulating LH/hCG receptor (LHCGR may identify pre-treatment IVF patients at risk of OHSS and poor implantation

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Chambers Anne E

2011-12-01

Full Text Available Abstract Background Successful pregnancy via in vitro fertilization (IVF depends on the recovery of an adequate number of healthy oocytes and on blastocyst implantation following uterine transfer. Two hormones, LH and hCG, utilize a common LH/hCG receptor (LHCGR, variations in which have profound implications in human reproduction. Soluble LHCGR (sLHCGR is released from experimental cell lines and placental explants and it can be detected in the follicular fluid and serum. Methods To evaluate the impact of circulating soluble LHCGR (sLHCGR in fertility treatment, we measured sLHCGR and LH-sLHCGR complex in serum from women seeking IVF using specifically developed quantitative enzyme-linked immunosorbent assays (ELISA. Following an IVF cycle of treatment, patients were grouped according to oocyte yield into low (lower than or equal to 7 oocytes, intermediate (8-14 oocytes and high (greater than or equal to 15 oocytes responders and pregnancy outcome noted. Results Pre-treatment sLHCGR identified many women at risk of ovarian hyperstimulation. Low levels of sLHCGR were associated with pregnancy in both high and low responders but sLHCGR did not significantly affect the treatment outcome of intermediate responders. Low responders who failed to become pregnant had high levels of circulating sLHCGR bound to LH (LH-sLHCGR. Conclusions Pre-treatment measurement of sLHCGR could be used to tailor individual fertility treatment programs and improve outcomes by avoiding ovarian hyperstimulation and poor embryo implantation.

Inhibition of Apoptosis Overcomes Stage-Related Compatibility Barriers to Chimera Formation in Mouse Embryos.

Science.gov (United States)

Masaki, Hideki; Kato-Itoh, Megumi; Takahashi, Yusuke; Umino, Ayumi; Sato, Hideyuki; Ito, Keiichi; Yanagida, Ayaka; Nishimura, Toshinobu; Yamaguchi, Tomoyuki; Hirabayashi, Masumi; Era, Takumi; Loh, Kyle M; Wu, Sean M; Weissman, Irving L; Nakauchi, Hiromitsu

2016-11-03

Cell types more advanced in development than embryonic stem cells, such as EpiSCs, fail to contribute to chimeras when injected into pre-implantation-stage blastocysts, apparently because the injected cells undergo apoptosis. Here we show that transient promotion of cell survival through expression of the anti-apoptotic gene BCL2 enables EpiSCs and Sox17 + endoderm progenitors to integrate into blastocysts and contribute to chimeric embryos. Upon injection into blastocyst, BCL2-expressing EpiSCs contributed to all bodily tissues in chimeric animals while Sox17 + endoderm progenitors specifically contributed in a region-specific fashion to endodermal tissues. In addition, BCL2 expression enabled rat EpiSCs to contribute to mouse embryonic chimeras, thereby forming interspecies chimeras that could survive to adulthood. Our system therefore provides a method to overcome cellular compatibility issues that typically restrict chimera formation. Application of this type of approach could broaden the use of embryonic chimeras, including region-specific chimeras, for basic developmental biology research and regenerative medicine. Copyright © 2016 Elsevier Inc. All rights reserved.

Outcome of Angiotensin II Inhibition in Pregnant Irradiated Rats and their Embryos

International Nuclear Information System (INIS)

Ramadan, F. L.; Ashry, Kh. M.

2010-01-01

The study aims to evaluate the synergism of losartan and or irradiation stress on the female rat mothers and their developing embryos as judged by the maternal biochemical pathways during gestation and teratogenic effects on the embryos. Losartan is an angiotensin II AT1-receptor antagonist used to regulate blood pressure. Losartan (5 mg/kg b.wt day) was daily orally administrated to pregnant rats from the 6 th to 18 th gestational days during which they were subjected to intermittent radiation dose levels of 0.5 Gy/4 times at the 9 th, 10 th, 11 th and 12 th days of gestation whereas investigation has been carried out one day prior to parturition. Dual treatment of losartan and radiation resulted in increased maternal serum levels of creatinine and bilirubin.The developing embryos in the uteri due to their high sensitivity showed various teratological, skeletal and histological impairment. Losartan and/or radiation induced effects were detected as growth retardation, malformations expressed as anopthalmia, kypophysis, subcutaneous haemorrhage and microtia as well as elevated intrauterium death and embryonic resorption. Moreover, the examination of endo skeletal system of fetuses showed retardation in the ossification of the skull bones and lack of ossification at the vertebrae and edges. Also, maternal and embryonic histological examination revealed that losartan and gamma radiation induced injury to kidney tissue manifested in rupture and shrinkage of renal corpuscle, infiltration, disappearance of glomularies, while the kidney of fetuses showed loss of renal pattern. Results point out that losartan should be used with caution in women at the reproductive age and those occupationally exposed to irradiation

HEALING OF MICROVENOUS PTFE PROSTHESES IMPLANTED INTO THE RAT FEMORAL VEIN

NARCIS (Netherlands)

VANDERLEI, B; DIJK, F; JONGEBLOED, WL; ROBINSON, PH; Bartels, H.

44 PTFE prostheses (Gore-Tex(R); ID 1 mm) were implanted into rats' femoral veins by means of the sleeve anastomotic technique and were evaluated at regular intervals from 1 h up till 24 weeks after implantation by means of light and electron microscopy to study in detail their healing process. All

Effects of 2.0 Gy of 60Co gamma rays irradiation on rat embryos

International Nuclear Information System (INIS)

Lee, Juing-Yi; Satow, Yukio

1987-01-01

Pregnant rats of Donryu strain were exposed to a whole-body 60 Co γ ray irradiation of a single dose of 2.0 Gy (Dose rate: 0.5 Gy/min) on day 7, 8, 9, 10 or 11 of gestation (sperm day = day 0). The rats were sacrificed on day 18 and the offspring were examined for external and visceral malformations. Malformed embryos occurred between days 7 and 11 with the highest incidence occurring on day 9. Dose with 2.0 Gy increased the rate of resorption or death (52.1 %), in the survivors, caused congenital malformation in a majority of embryos (86.5 %) on day 8 of gestation. There is an increase in malformation (93.3 %) and growth retardation, but no increase in mortality (42.9 %) on day 9 of gestation. Relatively few anomalies resulted from irradiation on day 7 of gestation. The peak day for cardiovascular anomalies occurred on day 9 (88.3 % of all survival embryos) with high levels also occurring on day 8 (86.5 %). Cardiovascular anomalies consisted of VSD, hypoplasia of the pulmonary trunk, coarctation of the aorta, double aortic arch, right aortic arch, riding aorta, complete transposition of the aorta, persistent atrioventricular canal, vascular ring, aberrant right subclavian artery and others. Similar anomalies, but at a lower incidence, were produced by 60 Co γ ray at dose levels of 2.0 Gy on day 10 or 11 of gestation. Cases of cleft lip and cleft palate or facial cleft were observed seventeen fetuses on day 9 of gestation (31 %). Exencephaly occurred in nine embryos treated on day 9 (16.1 %) and in one embryos treated on day 10. Tail defects appeared with treatment on day 9 with the latter predominating on day 11. The present study show that maximum resorption (52.1 %) was seen with treatment on day 8 whereas the highest rate of malformation (93.3 %) was observed with treatment on day 9. (J.P.N.)

Pre-implantation genetic screening using fluorescence in situ hybridization in couples of Indian ethnicity: Is there a scope?

Directory of Open Access Journals (Sweden)

Shailaja Gada Saxena

2014-01-01

Full Text Available Context: There is a high incidence of numerical chromosomal aberration in couples with repeated in vitro fertilization (IVF failure, advanced maternal age, repeated unexplained abortions, severe male factor infertility and unexplained infertility. Pre-implantation genetic screening (PGS, a variant of pre-implantation genetic diagnosis, screens numerical chromosomal aberrations in couples with normal karyotype, experiencing poor reproductive outcome. The present study includes the results of the initial pilot study on 9 couples who underwent 10 PGS cycles. Aim: The aim of the present study was to evaluate the beneficial effects of PGS in couples with poor reproductive outcome. Settings and Design: Data of initial 9 couples who underwent 10 PGS for various indications was evaluated. Subjects and Methods: Blastomere biopsy was performed on cleavage stage embryos and subjected to two round fluorescence in situ hybridization (FISH testing for chromosomes 13, 18, 21, X and Y as a two-step procedure. Results: Six of the 9 couples (10 PGS cycles conceived, including a twin pregnancy in a couple with male factor infertility, singleton pregnancies in a couple with secondary infertility, in three couples with adverse obstetric outcome in earlier pregnancies and in one couple with repeated IVF failure. Conclusion: In the absence of availability of array-comparative genomic hybridization in diagnostic clinical scenario for PGS and promising results with FISH based PGS as evident from the current pilot study, it is imperative to offer the best available services in the present scenario for better pregnancy outcome for patients.

Effects of pore size, implantation time, and nano-surface properties on rat skin ingrowth into percutaneous porous titanium implants.

Science.gov (United States)

Farrell, Brad J; Prilutsky, Boris I; Ritter, Jana M; Kelley, Sean; Popat, Ketul; Pitkin, Mark

2014-05-01

The main problem of percutaneous osseointegrated implants is poor skin-implant integration, which may cause infection. This study investigated the effects of pore size (Small, 40-100 μm and Large, 100-160 μm), nanotubular surface treatment (Nano), and duration of implantation (3 and 6 weeks) on skin ingrowth into porous titanium. Each implant type was percutaneously inserted in the back of 35 rats randomly assigned to seven groups. Implant extrusion rate was measured weekly and skin ingrowth into implants was determined histologically after harvesting implants. It was found that all three types of implants demonstrated skin tissue ingrowth of over 30% (at week 3) and 50% (at weeks 4-6) of total implant porous area under the skin; longer implantation resulted in greater skin ingrowth (p skin integration with the potential for a safe seal. Copyright © 2013 Wiley Periodicals, Inc.

Associations Between Pre-Implant Psychosocial Factors and Spinal Cord Stimulation Outcome: Evaluation Using the MMPI-2-RF.

Science.gov (United States)

Block, Andrew R; Marek, Ryan J; Ben-Porath, Yossef S; Kukal, Deborah

2017-01-01

Spinal cord stimulation (SCS) has variable effectiveness in controlling chronic pain. Previous research has demonstrated that psychosocial factors are associated with diminished results of SCS. The objective of this investigation is to examine associations between pre-implant psychological functioning as measured by the Minnesota Multiphasic Personality Inventory-2-Restructured Form (MMPI-2-RF) and SCS outcomes. SCS candidates at two sites (total N = 319) completed the MMPI-2-RF and measures of pain, emotional distress, and functional ability as part of a pre-implant psychological evaluation. At an average of 5 months post-implant, patients completed the measures of pain and emotional distress a second time. Poorer SCS outcomes and poorer patient satisfaction were associated with higher pre-implant MMPI-2-RF scores on scales used to assess emotional dysfunction, somatic/cognitive complaints, and interpersonal problems. Ways through which pre-implant psychological evaluations of spinal cord stimulator candidates can be informed by MMPI-2-RF findings are discussed. © The Author(s) 2015.

Effect of 935-MHz phone-simulating electromagnetic radiation on endometrial glandular cells during mouse embryo implantation.

Science.gov (United States)

Liu, Wenhui; Zheng, Xinmin; Qu, Zaiqing; Zhang, Ming; Zhou, Chun; Ma, Ling; Zhang, Yuanzhen

2012-10-01

This study examined the impact of 935MHz phone-simulating electromagnetic radiation on embryo implantation of pregnant mice. Each 7-week-old Kunming (KM) female white mouse was set up with a KM male mouse in a single cage for mating overnight after induction of ovulation. In the first three days of pregnancy, the pregnant mice was exposed to electromagnetic radiation at low-intensity (150 μW/cm(2), ranging from 130 to 200 μW/cm(2), for 2- or 4-h exposure every day), mid-intensity (570 μW/cm(2), ranging from 400 to 700 μW/cm(2), for 2- or 4-h exposure every day) or high-intensity (1400 μW/cm(2), ranging from 1200 to 1500 μW/cm(2), for 2- or 4-h exposure every day), respectively. On the day 4 after gestation (known as the window of murine embryo implantation), the endometrium was collected and the suspension of endometrial glandular cells was made. Laser scanning microscopy was employed to detect the mitochondrial membrane potential and intracellular calcium ion concentration. In high-intensity, 2- and 4-h groups, mitochondrial membrane potential of endometrial glandular cells was significantly lower than that in the normal control group (Pelectromagnetic radiation and longer length of the radiation are required to inflict a remarkable functional and structural damage to mitochondrial membrane. Our data demonstrated that electromagnetic radiation with a 935-MHz phone for 4 h conspicuously decreased mitochondrial membrane potential and lowered the calcium ion concentration of endometrial glandular cells. It is suggested that high-intensity electromagnetic radiation is very likely to induce the death of embryonic cells and decrease the chance of their implantation, thereby posing a high risk to pregnancy.

Influence of γ-irradiation on the structure and enzymatic activity of nuclear membrane in pregnant rats and their embryos

International Nuclear Information System (INIS)

Mirakhmedov, A.K.; Mirkhamidova, P.; Shamsutdinova, G.T.; Filatova, L.S.; Khamidov, D.Kh.; Zbarskij, I.B.; AN SSSR, Moscow

1992-01-01

Morphological and biochemical investigations of pregnant rats and embryo liver cell nuclei after in vivo irradiation in the doses of 1 and 2 Gy revealed their high radiosnsitivity at all stages of gestation and embryonal development. At damaging effect of radiation, we managed to observe sharp accumulation of products of lipid peroxide oxidation and suppresion of the activities of such enzymes in liver nuclei of pregnant rats and embryos. The changes of such a kind are shown to intensify with the increasing of irradiation doses. The most profound inhibition of activities of these enzymes in liver nuclei of embryos irradiated in utero was observed during the period of organogenesis (the 13th day of the development) and in fetal period of embryogenesis (the 17th day of the development), as well as the 13th and 17th day of gestation. The morphological data also demonstate the high level of cell nucleus sensitivity to the action of radiation during gestattion and embryogenesis

Live birth following serial vitrification of embryos and PGD for fragile X syndrome in a patient with the premutation and decreased ovarian reserve.

Science.gov (United States)

Nayot, Dan; Chung, Jin Tae; Son, Weon-Young; Ao, Assangla; Hughes, Mark; Dahan, Michael H

2013-11-01

To present a live birth resulting from serial vitrification of embryos and pre-implantation genetic diagnosis (PGD). A 31-year-old with primary infertility, fragile-X premutation, and decreased ovarian reserve (DOR) (baseline FSH level 33 IU/L), presented after failing to stimulate to follicle diameters >10 mm with three cycles of invitro fertilization (IVF). After counseling, the couple opted for serial in-vitro maturation (IVM), embryo vitrification, and genetic testing using array comparative genomic hybridization (aCGH) and PGD. Embryos were vitrified 2 days after intra-cytoplasmic sperm injection (ICSI). Thawed embryos were biopsied on day-three and transferred on day-five. The couple underwent 20 cycles of assisted reproductive technology. A total of 23 in-vivo mature and five immature oocytes were retrieved, of which one matured in-vitro. Of 24 embryos, 17/24 (71 %) developed to day two and 11/24 (46 %) survived to blastocyst stage with a biopsy result available. Four blastocysts had normal PGD and aCGH results. Both single embryo transfers resulted in a successful implantation, one a blighted ovum and the other in a live birth. Young patients with DOR have potential for live birth as long as oocytes can be obtained and embryos created. Serial vitrification may be the mechanism of choice in these patients when PGD is needed.

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

Science.gov (United States)

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

2012-10-01

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

[Association of human chorionic gonadotropin level in embryo culture media with early embryo development].

Science.gov (United States)

Wang, Haiying; Zhang, Renli; Han, Dong; Liu, Caixia; Cai, Jiajie; Bi, Yanling; Wen, Anmin; Quan, Song

2014-06-01

To investigate the association of human chorionic gonadotropin (HCG) level on day 3 of embryo culture with embryo development. Spent culture media were collected from individually cultured embryos on day 3 of in vitro fertilization and embryo transfer (IVF-ET) cycles. HCG concentration in the culture media was measured using an ELISA kit and its association with embryo development was assessed. In the 163 samples of embryo culture media from 60 patients, HCG was positive in 153 sample (93.8%) with a mean level of 0.85 ± 0.43 mIU/ml. The concentration of hCG in the culture media increased gradually as the number of blastomeres increased (F=2.273, P=0.03), and decreased as the morphological grade of the embryo was lowered (F=3.900, P=0.02). ELISA is capable of detecting HCG levels in spent culture media of embryos on day 3 of in vitro culture. The concentration of HCG in spent culture media is positively correlated with the status of early embryo development and implantation rate and thus serves as a useful marker for embryo selection in IVF-ET procedure.

The frequency of clinical pregnancy and implantation rate after cultivation of embryos in a medium with granulocyte macrophage colony-stimulating factor (GM-CSF) in patients with preceding failed attempts of ART.

Science.gov (United States)

Tevkin, S; Lokshin, V; Shishimorova, M; Polumiskov, V

2014-10-01

The application in IVF practice of modern techniques can improve positive outcome of each cycle in the assisted reproductive technology (ART) programs and the effectiveness of treatment as a whole. There are embryos in the female reproductive tract in physiological medium which contain various cytokines and growth factors. It plays an important role in the regulation of normal embryonic development, improve implantation and subsequently optimizing the development of the fetus and the placenta. Granulocyte macrophage colony-stimulating factor (GM-CSF is one of the cytokines playing an important role in reproductive function. Addition of recombinant GM-CSF to the culture medium can makes closer human embryos culture to in vivo conditions and improve the efficacy ART cycles. The analysis of culture embryos in EmbryoGen medium has shown that fertilization rate embryo culture and transfer to patients with previous unsuccessful attempts increases clinical pregnancy rate compared to the control group 39.1 versus 27.8%, respectively. It is noted that the implantation rate (on 7 weeks' gestation) and progressive clinical pregnancy rate (on 12 weeks' gestation) were significantly higher in group embryos culture in EmbryoGen medium compared to standard combination of medium (ISM1+VA), and were 20.4 and 17.4% versus 11.6 and 9.1%, respectively.

New Tools for Embryo Selection: Comprehensive Chromosome Screening by Array Comparative Genomic Hybridization

Directory of Open Access Journals (Sweden)

Lorena Rodrigo

2014-01-01

Full Text Available The objective of this study was to evaluate the usefulness of comprehensive chromosome screening (CCS using array comparative genomic hybridization (aCGH. The study included 1420 CCS cycles for recurrent miscarriage (n=203; repetitive implantation failure (n=188; severe male factor (n=116; previous trisomic pregnancy (n=33; and advanced maternal age (n=880. CCS was performed in cycles with fresh oocytes and embryos (n=774; mixed cycles with fresh and vitrified oocytes (n=320; mixed cycles with fresh and vitrified day-2 embryos (n=235; and mixed cycles with fresh and vitrified day-3 embryos (n=91. Day-3 embryo biopsy was performed and analyzed by aCGH followed by day-5 embryo transfer. Consistent implantation (range: 40.5–54.2% and pregnancy rates per transfer (range: 46.0–62.9% were obtained for all the indications and independently of the origin of the oocytes or embryos. However, a lower delivery rate per cycle was achieved in women aged over 40 years (18.1% due to the higher percentage of aneuploid embryos (85.3% and lower number of cycles with at least one euploid embryo available per transfer (40.3%. We concluded that aneuploidy is one of the major factors which affect embryo implantation.

Effects of ulipristal acetate on human embryo attachment and endometrial cell gene expression in an in vitro co-culture system.

Science.gov (United States)

Berger, C; Boggavarapu, N R; Menezes, J; Lalitkumar, P G L; Gemzell-Danielsson, K

2015-04-01

Does ulipristal acetate (UPA) used for emergency contraception (EC) interfere with the human embryo implantation process? UPA, at the dosage used for EC, does not affect human embryo implantation process, in vitro. A single pre-ovulatory dose of UPA (30 mg) acts by delaying or inhibiting ovulation and is recommended as first choice among emergency contraceptive pills due to its efficacy. The compound has also been demonstrated to have a dose-dependent effect on the endometrium, which theoretically could impair endometrial receptivity but its direct action on human embryo implantation has not yet been studied. Effect of UPA on embryo implantation process was studied in an in vitro endometrial construct. Human embryos were randomly added to the cultures and cultured for 5 more days with UPA (n = 10) or with vehicle alone (n = 10) to record the attachment of embryos. Endometrial biopsies were obtained from healthy, fertile women on cycle day LH+4 and stromal and epithelial cells were isolated. A three-dimensional in vitro endometrial co-culture system was constructed by mixing stromal cells with collagen covered with a layer of epithelial cells and cultured in progesterone containing medium until confluence. The treatment group received 200 ng/ml of UPA. Healthy, viable human embryos were placed on both control and treatment cultures. Five days later the cultures were tested for the attachment of embryos and the 3D endometrial constructs were analysed for endometrial receptivity markers by real-time PCR. There was no significant difference in the embryo attachment rate between the UPA treated group and the control group as 5 out of 10 human embryos exposed to UPA and 7 out of 10 embryos in the control group attached to the endometrial cell surface (P = 0.650). Out of 17 known receptivity genes studied here, only 2 genes, HBEGF (P = 0.009) and IL6 (P = 0.025) had a significant up-regulation and 4 genes, namely HAND2 (P = 0.003), OPN (P = 0.003), CALCR (P = 0.016) and

Improved ovulation rate and implantation in rats treated with royal jelly

African Journals Online (AJOL)

The ovaries and uteris of 12 mature female rats (Rattus norvegicus) were examined to determine the effect of commercial royal jelly on ovulation, ovarian weight and implantation rates. Rats were split in two groups of 6 each. Group one served as the treatment and group two the control. A daily dose of 25mg of royal jelly ...

Effects of 2. 0 Gy of /sup 60/Co gamma rays irradiation on rat embryos

Energy Technology Data Exchange (ETDEWEB)

Lee, Juing-Yi; Satow, Yukio

1987-01-01

Pregnant rats of Donryu strain were exposed to a whole-body /sup 60/Co ..gamma.. ray irradiation of a single dose of 2.0 Gy (Dose rate: 0.5 Gy/min) on day 7, 8, 9, 10 or 11 of gestation (sperm day = day 0). The rats were sacrificed on day 18 and the offspring were examined for external and visceral malformations. Malformed embryos occurred between days 7 and 11 with the highest incidence occurring on day 9. Dose with 2.0 Gy increased the rate of resorption or death (52.1 %), in the survivors, caused congenital malformation in a majority of embryos (86.5 %) on day 8 of gestation. There is an increase in malformation (93.3 %) and growth retardation, but no increase in mortality (42.9 %) on day 9 of gestation. Relatively few anomalies resulted from irradiation on day 7 of gestation. The peak day for cardiovascular anomalies occurred on day 9 (88.3 % of all survival embryos) with high levels also occurring on day 8 (86.5 %). Cardiovascular anomalies consisted of VSD, hypoplasia of the pulmonary trunk, coarctation of the aorta, double aortic arch, right aortic arch, riding aorta, complete transposition of the aorta, persistent atrioventricular canal, vascular ring, aberrant right subclavian artery and others. Similar anomalies, but at a lower incidence, were produced by /sup 60/Co ..gamma.. ray at dose levels of 2.0 Gy on day 10 or 11 of gestation. Cases of cleft lip and cleft palate or facial cleft were observed seventeen fetuses on day 9 of gestation (31 %). Exencephaly occurred in nine embryos treated on day 9 (16.1 %) and in one embryos treated on day 10. Tail defects appeared with treatment on day 9 with the latter predominating on day 11. The present study show that maximum resorption (52.1 %) was seen with treatment on day 8 whereas the highest rate of malformation (93.3 %) was observed with treatment on day 9. (J.P.N.).

Embryotoxic cytokines-Potential roles in embryo loss and fetal programming.

Science.gov (United States)

Robertson, Sarah A; Chin, Peck-Yin; Femia, Joseph G; Brown, Hannah M

2018-02-01

Cytokines in the reproductive tract environment at conception mediate a dialogue between the embryo and maternal tissues to profoundly influence embryo development and implantation success. Through effects on gene expression and the cell stress response, cytokines elicit an epigenetic impact with consequences for placental development and fetal growth, which in turn affect metabolic phenotype and long-term health of offspring. There is substantial evidence demonstrating that pro-survival cytokines, such as GM-CSF, CSF1, LIF, HB-EGF and IGFII, support embryos to develop optimally. Less attention has been paid to cytokines that adversely impact embryo development, including the pro-inflammatory cytokines TNF, TRAIL and IFNG. These agents elicit cell stress, impair cell survival and retard blastocyst development, and at sufficiently high concentrations, can cause embryo demise. Experiments in mice suggest these so-called 'embryotoxic' cytokines can harm embryos through pro-apoptotic and adverse programming effects, as well as indirectly suppressing uterine receptivity through the maternal immune response. Embryotrophic factors may mitigate against and protect from these adverse effects. Thus, the balance between embryotrophic and embryotoxic cytokines can impart effects on embryo development and implantation, and has the potential to contribute to endometrial 'biosensor' function to mediate embryo selection. Embryotoxic cytokines can be elevated in plasma and reproductive tract tissues in inflammatory conditions including infection, diabetes, obesity, PCOS and endometriosis. Studies are therefore warranted to investigate whether excessive embryotoxic cytokines contribute to infertility and recurrent implantation failure in women, and compromised reproductive performance in livestock animals. Copyright © 2018 Elsevier B.V. All rights reserved.

Trp53 deficient mice predisposed to preterm birth display region-specific lipid alterations at the embryo implantation site

Energy Technology Data Exchange (ETDEWEB)

Lanekoff, Ingela; Cha, Jeeyeon; Kyle, Jennifer E.; Dey, Sudhansu K.; Laskin, Julia; Burnum-Johnson, Kristin E.

2016-09-13

Here we demonstrate that conditional deletion of mouse uterine Trp53 (p53d/d), molecularly linked to mTORC1 activation and causally linked to premature uterine senescence and preterm birth, results in aberrant lipid signatures within the heterogeneous cell types of embryo implantation sites on day 8 of pregnancy. In situ nanospray desorption electrospray ionization mass spectrometry imaging (nano-DESI MSI) was used to characterize the molecular speciation of free fatty acids, monoacylglycerols, unmodified and oxidized phosphatidylcholine (PC/Ox-PC), and diacylglycerol (DG) species within implantation sites of p53d/d mice and floxed littermates. Implantation sites from p53d/d mice exhibited distinct spatially resolved changes demonstrating accumulation of DG species, depletion of Ox-PC species, and increase in species with more unsaturated acyl chains, including arachidonic and docosahexaenoic acid. Understanding abnormal changes in the abundance and localization of individual lipid species early in the progression to premature birth is important for discovering novel targets for treatments and diagnosis.

Effect of intrauterine position on the radiosensitivity of rat embryos

International Nuclear Information System (INIS)

Ward, W.F.; Aceto, H. Jr.; Karp, C.H.

1977-01-01

Rats were exposed to gamma rays or helium ions on one of days 4 to 9 of gestation. Embryonic survival was recorded as a function of intrauterine position at autopsy on day 20 of gestation. Embryos located at the ovarian and cervical ends of the uterus experienced higher rates of mortality than did their littermates located at the middle of the uterine horn. This effect was observed in litters exposed to both radiation modalities on all days studied. The influence of intrauterine position on embryonic survival was directly proportional to radiation dose and to the number of fetuses occupying the uterus horn. Under the least advantageous conditions (i.e., a crowded uterine horn exposed to a moderately high radiation dose), the cervical embryo's probability of survival was less than half that of the litter as a whole. A disproportionately high rate of embryonic mortality at the cervical position was also observed in litters irradiated under hypoxic conditions, suggesting that the non-random distribution of radiation effect was not the result of variations in oxygen concentration within the uterus. In contrast, there was no indication that intrauterine position influenced the distribution of gross morphologic abnormalities in irradiated litters

Mitochondrial DNA content in embryo culture medium is significantly associated with human embryo fragmentation.

Science.gov (United States)

Stigliani, S; Anserini, P; Venturini, P L; Scaruffi, P

2013-10-01

Is the amount of cell-free DNA released by human embryos into culture medium correlated with embryo morphological features? The mitochondrial DNA (mtDNA) content of culture medium is significantly associated with the fragmentation rate on Days 2 and 3 of embryo development, whether the oocyte came from women ≤ 35 or >35 years old. Cellular fragmentation is often utilized as one of the morphological parameters for embryo quality assessment. The amount of cellular fragments is considered to be an important morphological parameter for embryo implantation potential. It has been hypothesized that fragments are apoptotic bodies or anuclear cytoplasmatic pieces of blastomeres, although no definitive conclusion has been drawn about their pathogenesis. Human fertilized oocytes were individually cultured from Day 1 to Days 2 and 3. A total of 800 samples (166 spent media from Day 2 and 634 from Day 3) were enrolled into the present study. Double-stranded DNA (dsDNA) was quantified in 800 spent embryo culture media by Pico Green dye fluorescence assay. After DNA purification, genomic DNA (gDNA) and mtDNA were profiled by specific quantitative PCR. Statistical analyses defined correlations among DNA contents, embryo morphology and maternal age. Different independent tests confirmed the presence of DNA into embryo culture medium and, for the first time, we demonstrate that both gDNA and mtDNA are detectable in the secretome. The amount of DNA is larger in embryos with bad quality cleavage compared with high-grade embryos, suggesting that the DNA profile of culture medium is an objective marker for embryo quality assessment. In particular, DNA profiles are significantly associated with fragmentation feature (total dsDNA: P = 0.0010; mtDNA; P = 0.0247) and advanced maternal age. It is necessary to establish whether DNA profiling of spent embryo culture medium is a robust onsite test that can improve the prediction of blastulation, implantation and/or pregnancy rate. The

Vitamin E Phosphate Coating Stimulates Bone Deposition in Implant-related Infections in a Rat Model.

Science.gov (United States)

Lovati, Arianna B; Bottagisio, Marta; Maraldi, Susanna; Violatto, Martina B; Bortolin, Monica; De Vecchi, Elena; Bigini, Paolo; Drago, Lorenzo; Romanò, Carlo L

2018-06-01

Implant-related infections are associated with impaired bone healing and osseointegration. In vitro antiadhesive and antibacterial properties and in vivo antiinflammatory effects protecting against bone loss of various formulations of vitamin E have been demonstrated in animal models. However, to the best of our knowledge, no in vivo studies have demonstrated the synergistic activity of vitamin E in preventing bacterial adhesion to orthopaedic implants, thus supporting the bone-implant integration. The purpose of this study was to test whether a vitamin E phosphate coating on titanium implants may be able to reduce (1) the bacterial colonization of prosthetic implants and (2) bone resorption and osteomyelitis in a rat model of Staphylococcus aureus-induced implant-related infection. Twelve rats were bilaterally injected in the femurs with S aureus UAMS-1-Xen40 and implanted with uncoated or vitamin E phosphate-coated titanium Kirschner wires without local or systemic antibiotic prophylaxis. Eight rats represented the uninfected control group. A few hours after surgery, two control and three infected animals died as a result of unexpected complications. With the remaining rats, we assessed the presence of bacterial contamination with qualitative bioluminescence imaging and Gram-positive staining and with quantitative bacterial count. Bone changes in terms of resorption and osteomyelitis were quantitatively analyzed through micro-CT (bone mineral density) and semiquantitatively through histologic scoring systems. Six weeks after implantation, we found only a mild decrease in bacterial count in coated versus uncoated implants (Ti versus controls: mean difference [MD], -3.705; 95% confidence interval [CI], -4.416 to -2.994; p E-treated group compared with uncoated implants (knee joint: MD, -11.88; 95% CI, -16.100 to -7.664; p E-coated nails compared with the uncoated nails. These preliminary findings indicate that vitamin E phosphate implant coatings can exert a

A comparative study of zinc, magnesium, strontium-incorporated hydroxyapatite-coated titanium implants for osseointegration of osteopenic rats

Energy Technology Data Exchange (ETDEWEB)

Tao, Zhou-Shan [Department of Orthopaedic Surgery, Second Affiliated Hospital of Wenzhou Medical University, 109 Xueyuan Xi Road, Wenzhou, Zhejiang 325027 (China); Zhou, Wan-Shu [Endocrine & Metabolic Diseases Unit, Affiliated Hospital of Guizhou Medical University, Guizhou 550001 (China); He, Xing-Wen [Department of Orthopaedic Surgery, Hangzhou Bay Hospital of Ningbo, 315000 (China); Liu, Wei [Department of Orthopaedic Surgery, Jingmen No. 1 People' s Hospital, Jingmen 44800, Hubei (China); Bai, Bing-Li; Zhou, Qiang; Huang, Zheng-Liang; Tu, Kai-kai; Li, Hang; Sun, Tao [Department of Orthopaedic Surgery, Second Affiliated Hospital of Wenzhou Medical University, 109 Xueyuan Xi Road, Wenzhou, Zhejiang 325027 (China); Lv, Yang-Xun [Department of Orthopaedic Surgery, Wenzhou Central Hospital, Wenzhou, Zhejiang 325000 (China); Cui, Wei [Sichuan Provincial Orthopedics Hospital, No. 132 West First Section First Ring Road, Chengdu, Sichuan 610000 (China); Yang, Lei, E-mail: tzs19900327@163.com [Department of Orthopaedic Surgery, Second Affiliated Hospital of Wenzhou Medical University, 109 Xueyuan Xi Road, Wenzhou, Zhejiang 325027 (China)

2016-05-01

Surface modification techniques have been applied to generate titanium implant surfaces that promote osseointegration for the implants in cementless arthroplasty. However, its effect is not sufficient for osteoporotic bone. Zinc (Zn), magnesium (Mg), and strontium (Sr) present a beneficial effect on bone growth, and positively affect bone regeneration. The aim of this study was to confirm the different effects of the fixation strength of Zn, Mg, Sr-substituted hydroxyapatite-coated (Zn-HA-coated, Mg-HA-coated, Sr-HA-coated) titanium implants via electrochemical deposition in the osteoporotic condition. Female Sprague–Dawley rats were used for this study. Twelve weeks after bilateral ovariectomy, all animals were randomly divided into four groups: group HA; group Zn-HA; group Mg-HA and group Sr-HA. Afterwards, all rats from groups HA, Zn-HA, Mg-HA and Sr-HA received implants with hydroxyapatite containing 0%, 10% Zn ions, 10% Mg ions, and 10% Sr ions. Implants were inserted bilaterally in all animals until death at 12 weeks. The bilateral femurs of rats were harvested for evaluation. All treatment groups increased new bone formation around the surface of titanium rods and push-out force; group Sr-HA showed the strongest effects on new bone formation and biomechanical strength. Additionally, there are significant differences in bone formation and push-out force was observed between groups Zn-HA and Mg-HA. This finding suggests that Zn, Mg, Sr-substituted hydroxyapatite coatings can improve implant osseointegration, and the 10% Sr coating exhibited the best properties for implant osseointegration among the tested coatings in osteoporosis rats. - Highlights: • Surface modification techniques have been applied to generate titanium implant surfaces that promote osseointegration for the implants in cementless arthroplasty. • However, its effect is not sufficient for osteoporotic bone. Zinc (Zn), Magnesium(Mg), Strontium (Sr) present a benificial effect on bone

Obesity does not aggravate vitrification injury in mouse embryos: a prospective study

Directory of Open Access Journals (Sweden)

Ma Wenhong

2012-08-01

Full Text Available Abstract Background Obesity is associated with poor reproductive outcomes, but few reports have examined thawed embryo transfer in obese women. Many studies have shown that increased lipid accumulation aggravates vitrification injury in porcine and bovine embryos, but oocytes of these species have high lipid contents (63 ng and 161 ng, respectively. Almost nothing is known about lipids in human oocytes except that these cells are anecdotally known to be relatively lipid poor. In this regard, human oocytes are considered to be similar to those of the mouse, which contain approximately 4 ng total lipids/oocyte. To date, no available data show the impact of obesity on vitrification in mouse embryos. The aim of this study was to establish a murine model of maternal diet-induced obesity and to characterize the effect of obesity on vitrification by investigating the survival rate and embryo developmental competence after thawing. Methods Prospective comparisons were performed between six–eight-cell embryos from obese and normal-weight mice and between fresh and vitrified embryos. Female C57BL/6 mice were fed standard rodent chow (normal-weight group or a high-fat diet (obese group for 6 weeks. The mice were mated, zygotes were collected from oviducts and cultured for 3 days, and six–eight-cell embryos were then selected to assess lipid content in fresh embryos and to evaluate differences in apoptosis, survival, and development rates in response to vitrification. Results In fresh embryos from obese mice, the lipid content (0.044 vs 0.030, Pvs.9.3%, Pvs. 93.1%, P Conclusions This study demonstrated that differences in survival and developmental rates between embryos from obese and normal-weight mice were eliminated after vitrification. Thus, maternal obesity does not aggravate vitrification injury, but obesity alone greatly impairs pre-implantation embryo survival and development.

Human embryo-conditioned medium stimulates in vitro endometrial angiogenesis

NARCIS (Netherlands)

Kapiteijn, K.; Koolwijk, P.; Weiden, R.M.F. van der; Nieuw Amerongen, G. van; Plaisier, M.; Hinsbergh, V.W.M. van; Helmerhorst, F.M.

2006-01-01

Objective: Successful implantation and placentation depend on the interaction between the endometrium and the embryo. Angiogenesis is crucial at this time. In this article we investigate the direct influence of the human embryo on in vitro endometrial angiogenesis. Design: In vitro study. Setting:

[How can we nowadays select the best embryo to transfer?].

Science.gov (United States)

Alter, L; Boitrelle, F; Sifer, C

2014-01-01

Multiple pregnancies stand as the most common adverse outcome of assisted reproduction technologies (ART) and the dangers associated with those pregnancies have been reduced by doing elective single embryo transfers (e-SET). Many studies have shown that e-SET is compatible with a continuously high pregnancy rate per embryo transfer. Yet, it still becomes necessary to improve the selection process in order to define the quality of individual embryos - so that the ones we choose for transfer are more likely to implant. First, analysis of embryo morphology has greatly helped in this identification and remains the most relevant criterion for choosing the embryo. The introduction of time-lapse imaging provides new criteria predictive of implantation potential, but the real contribution of this system - including the benefit/cost ratio - seems to be not yet properly established. In this context, extended culture until blastocyst stage is an essential practice but it appears wise to keep it for a population showing a good prognosis. Then, the failure of aneuploid embryos to implant properly led to achieve preimplantation genetic screening (PGS) in order to increase pregnancy and delivery rates after ART. However, PGS by fluorescence in situ hybridization (FISH) at day 3 is a useless process - and may even be harmful. Another solution involves using comparative genomic hybridisation (CGH) and moving to blastocyst biopsy. Finally, it is envisaged that morphology will also be significantly aided by non-invasive analysis of biomarkers in the culture media that give a better reflection of whole-embryo physiology and function. Copyright © 2014. Published by Elsevier SAS.

Freezing of Rat Tibiae at -20°C Does Not Affect the Mechanical Properties of Intramedullary Bone/Implant-Interface: Brief Report

Science.gov (United States)

Diefenbeck, Michael; Mückley, Thomas; Zankovych, Sergiy; Bossert, Jörg; Jandt, Klaus D; Schrader, Christian; Schmidt, Jürgen; Finger, Ulrich; Faucon, Mathilde

2011-01-01

Background: The effects of freezing-thawing cycles on intramedullary bone-implant interfaces have been studied in a rat model in mechanical pull-out tests. Implants: Twenty TiAl6V4 rods (Ø 0.8 mm, length 10 mm) implanted in rat tibiae Methods: 10 rats underwent bilateral tibial implantation of titanium rods. At eight weeks, the animals were sacrificed and tibiae harvested for biomechanical testing. Eight tibiae were frozen and stored at -20°C for 14 days, the remaining eight were evaluated immediately post-harvest. Pull-out tests were used to determine maximum force and interfacial shear strength. Results: There were no significant differences between fresh and those of the frozen-thawed group in maximum force or in interfacial shear strength. Conclusion: Frozen Storage of rat tibiae containing implants at -20° C has no effects on the biomechanical properties of Bone/ Implant interface. PMID:21760868

Effect of implantation of biodegradable magnesium alloy on BMP-2 expression in bone of ovariectomized osteoporosis rats

International Nuclear Information System (INIS)

Guo, Yue; Ren, Ling; Liu, Chang; Yuan, Yajiang; Lin, Xiao; Tan, Lili; Chen, Shurui; Yang, Ke; Mei, Xifan

2013-01-01

The study was focused on the implantation of a biodegradable AZ31 magnesium alloy into the femoral periosteal of the osteoporosis modeled rats. The experimental results showed that after 4 weeks implantation of AZ31 alloy in the osteoporosis modeled rats, the expression of BMP-2 in bone tissues of the rats was much enhanced, even higher than the control group, which should promote the bone formation and be beneficial for reducing the harmful effect of osteoporosis. Results of HE stains showed that the implantation of AZ31 alloy did not have obvious pathological changes on both the liver and kidney of the animal. - Highlights: • Mg alloy greatly increased expression of BMP-2 in osteoporosis modeled rat bone. • Mg alloy showed good biological safety. • Mg alloy is beneficial for reducing the symptom of osteoporosis

Embryo disposition and the new death scene

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Ellison, David

2011-01-01

Full Text Available In the IVF clinic - a place designed principally for the production and implantation of embryos - scientists and IVF recipients are faced with decisions regarding the disposition of frozen embryos. At this time there are hundred of thousands of cryopreserved embryos awaiting such determinations. They may be thawed for transfer to the woman herself, they may be donated for research or for use by other infertile couples, they may remain in frozen storage, or they may variously be discarded by being allowed to 'succumb', or 'perish'. Where the choice is discard, some IVF clients have chosen to formalise the process through ceremony. A new language is emerging in response to the desires of the would-be-parents who might wish to characterise the discard experience as a ‘good death’. This article examines the procedure known as ‘compassionate transfer’ where the embryo to be discarded is placed in the woman’s vagina where it is clear that it will not develop further. An alternate method has the embryo transferred in the usual manner but without the benefit of fertility-enhancing hormones at a point in the cycle unreceptive to implantation. The embryo destined for disposal is thus removed from the realm of technological possibility and ‘returned’ to the female body for a homely death. While debates continue about whether or not embryos constitute life, new practices are developing in response to the emotional experience of embryo discard. We argue that compassionate transfer is a death scene taking shape. In this article, we take the measure of this new death scene’s fabrication, and consider the form, significance, and legal complexity of its ceremonies.

Optimization of cell adhesion on mg based implant materials by pre-incubation under cell culture conditions.

Science.gov (United States)

Willumeit, Regine; Möhring, Anneke; Feyerabend, Frank

2014-05-05

Magnesium based implants could revolutionize applications where orthopedic implants such as nails, screws or bone plates are used because they are load bearing and degrade over time. This prevents a second surgery to remove conventional implants. To improve the biocompatibility we studied here if and for how long a pre-incubation of the material under cell culture conditions is favorable for cell attachment and proliferation. For two materials, Mg and Mg10Gd1Nd, we could show that 6 h pre-incubation are already enough to form a natural protective layer suitable for cell culture.

Optimization of Cell Adhesion on Mg Based Implant Materials by Pre-Incubation under Cell Culture Conditions

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Regine Willumeit

2014-05-01

Full Text Available Magnesium based implants could revolutionize applications where orthopedic implants such as nails, screws or bone plates are used because they are load bearing and degrade over time. This prevents a second surgery to remove conventional implants. To improve the biocompatibility we studied here if and for how long a pre-incubation of the material under cell culture conditions is favorable for cell attachment and proliferation. For two materials, Mg and Mg10Gd1Nd, we could show that 6 h pre-incubation are already enough to form a natural protective layer suitable for cell culture.

Effects of the Oral Oxytocin Receptor Antagonist Tocolytic OBE001 on Reproduction in Rats.

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Pohl, Oliver; Perks, Deborah; Rhodes, Jon; Comotto, Laura; Baldrick, Paul; Chollet, André

2016-04-01

OBE001 is a novel, orally active nonpeptide oxytocin receptor antagonist under development for the treatment of preterm labor and improvement in embryo implantation and pregnancy rate in assisted reproductive technology (ART). The reproductive safety of OBE001 was evaluated in customized fertility embryonic development (FER)/early embryonic development (EED) and fetal development (FD) and pre/postnatal development (PPN) studies mimicking clinical exposure scenarios. Oral OBE001 was evaluated at doses of 37.5, 75, and 125 mg/kg/d in female rats during a FER/EED study (from premating to implantation) and throughout FD during a FD/PPN study. No OBE001 effects were observed during the FER/EED study. The FD/PPN study did not result in adverse OBE001 effects in females allowed to litter, their offspring, and second-generation fetuses. Females at 125 mg/kg/d who underwent cesarean section before term had slight reductions in body weights and food consumption, and associated fetuses had slightly delayed ossification of skull bones, which was not adverse in the absence of effects on live offspring. OBE001 at up to 125 mg/kg/d had no effects on EED and no adverse effects on FD and postnatal development of rats. These results constitute an important step toward the development of OBE001 in preterm labor and ART indications. © The Author(s) 2015.

Glucose uptake and transport in contracting, perfused rat muscle with different pre-contraction glycogen concentrations

DEFF Research Database (Denmark)

Hespel, P; Richter, Erik

1990-01-01

1. Glucose uptake and transport, muscle glycogen, free glucose and glucose-6-phosphate concentrations were studied in perfused resting and contracting rat skeletal muscle with different pre-contraction glycogen concentrations. Rats were pre-conditioned by a combination of swimming exercise and diet......, resulting in either low (glycogen-depleted rats), normal (control rats) or high (supercompensated rats) muscle glycogen concentrations at the time their hindlimbs were perfused. 2. Compared with control rats, pre-contraction muscle glycogen concentration was approximately 40% lower in glycogen-depleted rats......, whereas it was 40% higher in supercompensated rats. Muscle glycogen break-down correlated positively (r = 0.76; P less than 0.001) with pre-contraction muscle glycogen concentration. 3. Glucose uptake during contractions was approximately 50% higher in glycogen-depleted hindquarters than in control...

Approaches for prediction of the implantation potential of human embryos

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Georgi Stamenov

2013-01-01

Full Text Available Optimization of assisted reproductive technologies (ART has become the main goal of contemporary reproductive medicine. The main aspiration of scientists working in the field is to use less intervention to achieve more, and, if possible, in a more cost-effective way. A number of directions have been under development, namely – various stimulation protocols, ART with no stimulation whatever, all aiming at a single goal – the chase for Moby Dick, or the perfect embryo. Comprehensive embryo selection resulting in reducing the number of transferred embryos is one of the main directions for optimization of the ART procedures. Both clinical and laboratory procedures are being constantly improved, and today there is a significant number of clinics that report success rates of 30% and even higher. Based on results achieved, and analyzing data from millions of ART procedures, researchers from different centers are seeking to develop prognostic models in order to further improve success rates. One of the greatest challenges remains the reduction of the incidence of multifetal pregnancy, and that can be achieved only through reducing the number of embryos per transfer and a rise in single embryo transfer (SET numbers. This, however, depends on reliable methods for preliminary embryo selection, employing a growing number of morphological, biochemical, genetic and other characteristics of the embryo. A primary concern in developing prognostic models for in vitro fertilization (IVF outcome is selecting the prognostic parameters to be included. A number of publications define the main criteria that have an impact on fertilization outcome on the side of the embryo, and for the ultimate outcome of the ART procedure – on the side of the maternal organism as a whole. In this review, some of the most important parameters are discussed, with particular focus on their application for development of IVF prognostic models.

Induction of dominant lethals in male mice treated as embryos with 35S

International Nuclear Information System (INIS)

Reddy, K.S.; Reddy, P.P.; Reddy, O.S.

1980-01-01

Pregnant female mice were injected (ip) with 20 μCi of 35 S or 0.5 ml of saline (control) on 3.5 day of gestation. The animals were allowed to litter and the (CBA female x C 3 H/He male) F 1 males treated as embryos were tested at maturity (8-10 weeks) for dominant lethal incidence. Each male was mated to 3 untreated virgin females for a period of 3 weeks. The pregnant animals were killed at mid gestation and the uterine contents and corpora lutea were examined. There was a significant increase in the frequency of dominant lethals both at pre- and post-implantation stages in the treated group when compared to controls. As a result a significant increase in dead implantations/female and reduction in live implantations/female were noticed in the treated group. Thus the results clearly delineate the genetic effects of sulfur-35 in mice. (auth.)

Evaluation of porous vitreous carbon or silicon implants by radiology in rat's skull; Avaliacao radiologica de implantes de carbono vitreo poroso ou silicone em cranio de ratos

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Vaccari-Mazzetti, Marcelo Paulo; Kobata, Celio Toshiro [Lusiada University of Santos, SP (Brazil). Hospital Defeitos da Face. Dept. of Surgery]. E-mail: mmgvaccari@ig.com.br; Fabiani, Paulo [Lusiada University of Santos, SP (Brazil). Dept. of Radiology; Martins, Dulce Maria Fonseca Soares [Universidade Federal de Sao Paulo (UNIFESP/EPM), SP (Brazil). Dept. of Surgery. Div. of Plastic Surgery; Gomes, Paulo de Oliveira [Universidade Federal de Sao Paulo (UNIFESP/EPM), SP (Brazil). Dept. of Surgery. Div. of Operatory Technique and Experimental Surgery; Martins, Jose Luiz [Universidade Federal de Sao Paulo (UNIFESP/EPM), SP (Brazil). Dept. of Surgery. Div. of Pediatric Surgery

2008-07-01

Purpose: Evaluate by CT the use of porous vitreous carbon (PVC) and silicon (S) implants as the replacement bone in the craniofacial skeleton of rats. Methods: 40 rats divided in: Group A (n=20) PVC submitted to the implant of a fragment in skull. After the euthanasia, the animals were divided into two subgroups: A I: 10 animals, studied in the 7th postoperative day (P.O) and AII: 10 animals, studied in the 28th P.O. In group B, S, 20 rats were submitted to S implant in the skull. All other steps were identical to group A, with designation of subgroups BI and BII. CT with beams in axial cuts of 1 mm thickness to obtain 3-D information It was used Hounsfield scale for evaluate the radio density of the implant. They were used non parametric tests to analyze the results. Results: The 7th PO boss remained in the two groups, but for 28th PO, observed reduction in the volume of the implant in Group A, not observed in group B. CT studies noticed different radio densities around all of S prostheses (pseudo capsule), that do not appeared in CPV implants. The S has remained unchanged in the CT, but the CPV has had a modification in its radio density (p{<=}0,05), in all implants. Conclusion: In CT evaluation the implants of CPV have greater deformation that the S, which makes them not suitable for replacement of membranous bone in the rat skull. (author)

Rat embryo fibroblasts require both the cell-binding and the heparin-binding domains of fibronectin for survival

DEFF Research Database (Denmark)

Jeong, J; Han, I; Lim, Y

2001-01-01

of the cell-binding domain of FN with integrin is sufficient to rescue rat embryo fibroblasts (REFs) from detachment-induced apoptosis. REFs attached and spread normally after plating on substrates coated with either intact FN or a FN fragment, FN120, that contains the cell-binding domain but lacks the C...

Pre-differentiation of mesenchymal stromal cells in combination with a microstructured nerve guide supports peripheral nerve regeneration in the rat sciatic nerve model.

Science.gov (United States)

Boecker, Arne Hendrik; van Neerven, Sabien Geraldine Antonia; Scheffel, Juliane; Tank, Julian; Altinova, Haktan; Seidensticker, Katrin; Deumens, Ronald; Tolba, Rene; Weis, Joachim; Brook, Gary Anthony; Pallua, Norbert; Bozkurt, Ahmet

2016-02-01

Many bioartificial nerve guides have been investigated pre-clinically for their nerve regeneration-supporting function, often in comparison to autologous nerve transplantation, which is still regarded as the current clinical gold standard. Enrichment of these scaffolds with cells intended to support axonal regeneration has been explored as a strategy to boost axonal regeneration across these nerve guides Ansselin et al. (1998). In the present study, 20 mm rat sciatic nerve defects were implanted with a cell-seeded microstructured collagen nerve guide (Perimaix) or an autologous nerve graft. Under the influence of seeded, pre-differentiated mesenchymal stromal cells, axons regenerated well into the Perimaix nerve guide. Myelination-related parameters, like myelin sheath thickness, benefitted from an additional seeding with pre-differentiated mesenchymal stromal cells. Furthermore, both the number of retrogradely labelled sensory neurons and the axon density within the implant were elevated in the cell-seeded scaffold group with pre-differentiated mesenchymal stromal cells. However, a pre-differentiation had no influence on functional recovery. An additional cell seeding of the Perimaix nerve guide with mesenchymal stromal cells led to an extent of functional recovery, independent of the differentiation status, similar to autologous nerve transplantation. These findings encourage further investigations on pre-differentiated mesenchymal stromal cells as a cellular support for peripheral nerve regeneration. © 2015 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

Endometrial injury to overcome recurrent embryo implantation failure: a systematic review and meta-analysis.

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Potdar, Neelam; Gelbaya, Tarek; Nardo, Luciano G

2012-12-01

Mechanical endometrial injury (biopsy/scratch or hysteroscopy) in the cycle preceding ovarian stimulation for IVF has been proposed to improve implantation in women with unexplained recurrent implantation failure (RIF). This is a systematic review and meta-analysis of studies comparing the efficacy of endometrial injury versus no intervention in women with RIF undergoing IVF. All controlled studies of endometrial biopsy/scratch or hysteroscopy performed in the cycle preceding ovarian stimulation were included and the primary outcome measure was clinical pregnancy rate. Pooling of seven controlled studies (four randomized and three non-randomized), with 2062 participants, showed that local endometrial injury induced in the cycle preceding ovarian stimulation is 70% more likely to result in a clinical pregnancy as opposed to no intervention. There was no statistically significant heterogeneity in the methods used, clinical pregnancy rates being twice as high with biopsy/scratch (RR 2.32, 95% CI 1.72-3.13) as opposed to hysteroscopy (RR 1.51, 95% CI 1.30-1.75). The evidence is strongly in favour of inducing local endometrial injury in the preceding cycle of ovarian stimulation to improve pregnancy outcomes in women with unexplained RIF. However, large randomized studies are required before iatrogenic induction of local endometrial injury can be warranted in routine clinical practice. Some women undergoing IVF treatment fail to conceive despite several attempts with good-quality embryos and no identifiable reason. We call this 'recurrent implantation failure' (RIF) where the embryo fails to embed or implant within the lining of the womb. Studies have shown that inducing injury to the lining of the womb in the cycle before starting ovarian stimulation for IVF can help improve the chances of achieving pregnancy. Injury can be induced by either scratching the lining of the womb using a biopsy tube or by telescopic investigation of the womb using a camera. We performed a

The impact of preimplantation genetic diagnosis on human embryos

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García-Ferreyra J.

2016-12-01

Full Text Available Chromosome abnormalities are extremely common in human oocytes and embryos and are associated with a variety of negative outcomes for both natural cycles and those using assisted reproduction techniques. Aneuploidies embryos may fail to implant in the uterus, miscarry, or lead to children with serious medical problems (e.g., Down syndrome. Preimplantation genetic diagnosis (PGD is a technique that allows the detection of aneuploidy in embryos and seeks to improve the clinical outcomes od assisted reproduction treatments, by ensuring that the embryos chosen for the transfer are chromosomally normal.

Experimental rat lung tumor model with intrabronchial tumor cell implantation.

Science.gov (United States)

Gomes Neto, Antero; Simão, Antônio Felipe Leite; Miranda, Samuel de Paula; Mourão, Lívia Talita Cajaseiras; Bezerra, Nilfácio Prado; Almeida, Paulo Roberto Carvalho de; Ribeiro, Ronaldo de Albuquerque

2008-01-01

The objective of this study was to develop a rat lung tumor model for anticancer drug testing. Sixty-two female Wistar rats weighing 208 +/- 20 g were anesthetized intraperitoneally with 2.5% tribromoethanol (1 ml/100 g live weight), tracheotomized and intubated with an ultrafine catheter for inoculation with Walker's tumor cells. In the first step of the experiment, a technique was established for intrabronchial implantation of 10(5) to 5 x 10(5) tumor cells, and the tumor take rate was determined. The second stage consisted of determining tumor volume, correlating findings from high-resolution computed tomography (HRCT) with findings from necropsia and determining time of survival. The tumor take rate was 94.7% for implants with 4 x 10(5) tumor cells, HRCT and necropsia findings matched closely (r=0.953; p<0.0001), the median time of survival was 11 days, and surgical mortality was 4.8%. The present rat lung tumor model was shown to be feasible: the take rate was high, surgical mortality was negligible and the procedure was simple to perform and easily reproduced. HRCT was found to be a highly accurate tool for tumor diagnosis, localization and measurement and may be recommended for monitoring tumor growth in this model.

Elevated expression of proto-oncogenes accompany enhanced induction of heat-shock genes after exposure of rat embryos in utero to ionizing irradiation

International Nuclear Information System (INIS)

Higo, H.; Lee, J.Y.; Satow, Y.; Higo, K.

1989-01-01

We have recently found that the effects of exposing rat embryos in utero to teratogens capable of producing cardiac anomalies were expressed later as enhanced induction of heat-shock proteins (hsp70 family) when embryonic hearts were cultured in vitro. However, it remained to be determined whether heat-shock proteins are induced in vivo after exposure to teratogens. The heat-shock response in some mammalian systems is known to be accompanied by elevated expression of proto-oncogenes. Using gene-specific DNA probes, we examined the levels of the expression (transcription) of heat-shock protein genes and two nuclear proto-oncogenes, c-fos and c-myc, in the embryos removed from irradiated pregnant mother rats 4 or 5 days after the irradiation. We found that the levels of expression in vivo of the hsp70 and c-myc genes in the irradiated embryos increased by approximately twofold as compared with those in the control. The expression in vivo of the c-fos gene was not detected in either the irradiated or non-irradiated embryos. After 0.5-hr incubation in vitro of the embryos, however, the expression of the c-fos gene in the irradiated embryos was highly enhanced whereas the control showed no changes. Although the exact functions of these gene products still remain obscure, the enhanced expression of hsp70 gene(s) and the nuclear proto-oncogenes observed in the present study may reflect repair of intracellular damages and/or regeneration of tissue by compensatory cell proliferation, processes that may disturb the normal program of organogenesis

Implantation of Octacalcium Phosphate Stimulates both Chondrogenesis and Osteogenesis in the Tibia, but Only Osteogenesis in the Rat Mandible

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F. Sargolzaei Aval

2006-09-01

Full Text Available Statement of problem: It is not known whether endochondral and intramembranous bones have distinct biological characteristics. Octacalcium Phosphate (OCP, a hydroxyapatite precursor, has been reported to stimulate bone formation after being implanted in parietal bone defects of rats.Purpose: The present study was designed to investigate the response of endochondral and intramembranous bones to OCP implantation and to compare their biological characteristicsMaterials and Methods: Full-thickness standardized trephine defects were made in rat tibiae and mandibles and synthetic OCP was implanted into the defects. The biologic response was examined histologically to identify bone and cartilage formation.Results: Both chondrogenesis and osteogenesis were initiated in the tibia, 1 week after implantation of OCP and most of the cartilage was replaced by bone at week 2.However, the mandible only showed osteogenesis in response to OCP implantation at week 2, and no cartilage formation was associated with the osteogenesis.Conclusions: According to the results obtained in the present study, endochondral and intramembranous bones exhibit different biological responses to OCP implantation in rats.

Pre-, intra- and post-operative imaging of cochlear implants

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Vogl, T.J.; Naguib, N.N.N.; Burck, I. [University Hospital Frankfurt (Germany). Inst. of Diagnostic and Interventional Radiology; Tawfik, A. [Mansoura Univ. (Egypt). Dept. of Diagnostic and Interventional Radiology; Emam, A. [University Hospital Alexandria (Egypt). Dept. of Diagnostic and Interventional Radiology; Nour-Eldin, A. [University Hospital Cairo (Egypt). Dept. of Radiology; Stoever, T. [University Hospital of Frankfurt (Germany). Dept. of Otolaryngology

2015-11-15

The purpose of this review is to present essential imaging aspects in patients who are candidates for a possible cochlear implant as well as in postsurgical follow-up. Imaging plays a major role in providing information on preinterventional topography, variations and possible infections. Preoperative imaging using DVT, CT, MRI or CT and MRI together is essential for candidate selection, planning of surgical approach and exclusion of contraindications like the complete absence of the cochlea or cochlear nerve, or infection. Relative contraindications are variations of the cochlea and vestibulum. Intraoperative imaging can be performed by fluoroscopy, mobile radiography or DVT. Postoperative imaging is regularly performed by conventional X-ray, DVT, or CT. In summary, radiological imaging has its essential role in the pre- and post-interventional period for patients who are candidates for cochlear implants.

Cannabis sativa smoke inhalation decreases bone filling around titanium implants: a histomorphometric study in rats.

Science.gov (United States)

Nogueira-Filho, Getulio da R; Cadide, Tiago; Rosa, Bruno T; Neiva, Tiago G; Tunes, Roberto; Peruzzo, Daiane; Nociti, Francisco Humberto; César-Neto, João B

2008-12-01

Although the harmful effect of tobacco smoking on titanium implants has been documented, no studies have investigated the effects of cannabis sativa (marijuana) smoking. Thus, this study investigated whether marijuana smoke influences bone healing around titanium implants. Thirty Wistar rats were used. After anesthesia, the tibiae surface was exposed and 1 screw-shaped titanium implant was placed bilaterally. The animals were randomly assigned to one of the following groups: control (n = 15) and marijuana smoke inhalation (MSI) 8 min/d (n = 15). Urine samples were obtained to detect the presence of tetra-hidro-cannabinoid. After 60 days, the animals were killed. The degree of bone-to-implant contact and the bone area within the limits of the threads of the implant were measured in the cortical (zone A) and cancellous bone (zone B). Tetra-hidro-cannabinoid in urine was positive only for the rats of MSI group. Intergroup analysis did not indicate differences in zone A-cortical bone (P > 0.01), however, a negative effect of marijuana smoke (MSI group) was observed in zone B-cancellous bone for bone-to-implant contact and bone area (Student's t test, P smoke on bone healing may represent a new concern for implant success/failure.

Potent radio-protective effects of vitamins E and C on radiation induced DNA damage in gametes leading to lower frequencies of chromosomal aberrations and micronuclei in subsequent embryos

International Nuclear Information System (INIS)

Hossein Mozdarani

2007-01-01

Complete text of publication follows. Objective: To compare the effects of parental and maternal exposure of NMRI mice with γ-rays on gametes in the absence or presence of vitamins E and C and subsequent cytogenetic damage in pre-implantation embryos generated from irradiated gametes. Materials and Methods: Male and female NMRI mice were whole body irradiated in the presence of 200 IU/Kg vitamin E and 100 μg/ml vitamin C. Various mating schemes were designed for mating of irradiated mice, e.g. mating irradiated male with non-irradiated female, irradiated female with non irradiated male or both male and female irradiated. About 68 h post coitus, 4-8-cell embryos were flushed out from oviducts and fixed on slides using standard methods in order to screen for chromosome abnormalities and micronuclei. Results: In control embryos, frequencies of abnormal metaphase and embryos with micronuclei was low and there was no significant difference between vitamins treated samples and controls. However there was an increase in both abnormal metaphases and micronuclei frequency in embryos generated after parental or maternal irradiation or both. Vitamin E effectively reduced the frequency of aneuploidy in all irradiated groups and vitamin C was very effective in reducing the frequencies of micronuclei. DRF calculated for both vitamins indicate that vitamin C is more potent than vitamin E in reducing clastogenic effects of gamma-rays in pre-implantation embryos. Conclusion: Data indicate that γ-irradiation affects spermatogenesis and preovulatory stage oocytes in male and female mice respectively. These effects might be due to DNA alterations in sperms and oocytes affecting meiotic segregations that may lead to chromosome abnormalities in subsequent embryos expressed as numerical chromosome abnormalities or micronuclei. Administration of vitamins E and C before irradiation effectively reduced the frequency of chromosomal abnormalities. The way these vitamins reduces genotoxic

The methyltransferase Setdb1 is essential for meiosis and mitosis in mouse oocytes and early embryos.

Science.gov (United States)

Eymery, Angeline; Liu, Zichuan; Ozonov, Evgeniy A; Stadler, Michael B; Peters, Antoine H F M

2016-08-01

Oocytes develop the competence for meiosis and early embryogenesis during their growth. Setdb1 is a histone H3 lysine 9 (H3K9) methyltransferase required for post-implantation development and has been implicated in the transcriptional silencing of genes and endogenous retroviral elements (ERVs). To address its role in oogenesis and pre-implantation development, we conditionally deleted Setdb1 in growing oocytes. Loss of Setdb1 expression greatly impaired meiosis. It delayed meiotic resumption, altered the dynamics of chromatin condensation, and impaired kinetochore-spindle interactions, bipolar spindle organization and chromosome segregation in more mature oocytes. The observed phenotypes related to changes in abundance of specific transcripts in mutant oocytes. Setdb1 maternally deficient embryos arrested during pre-implantation development and showed comparable defects during cell cycle progression and in chromosome segregation. Finally, transcriptional profiling data indicate that Setdb1 downregulates rather than silences expression of ERVK and ERVL-MaLR retrotransposons and associated chimearic transcripts during oogenesis. Our results identify Setdb1 as a newly discovered meiotic and embryonic competence factor safeguarding genome integrity at the onset of life. © 2016. Published by The Company of Biologists Ltd.

Live births after polar body biopsy and frozen-thawed cleavage stage embryo transfer: case report.

Science.gov (United States)

Guimarães, Fernando; Roque, Matheus; Valle, Marcello; Kostolias, Alessandra; Azevedo, Rodrigo A de; Martinhago, Ciro D; Sampaio, Marcos; Geber, Selmo

2016-12-01

Pre-implantation genetic diagnosis (PGD) or screening (PGS) technology, has emerged and developed in the past few years, benefiting couples as it allows the selection and transfer of healthy embryos during IVF treatments. These techniques can be performed in oocytes (polar-body biopsy) or embryos (blastomere or trophectoderm biopsy). In this case report, we describe the first two live births to be published in Brazil after a polar-body (PB) biopsy. In case 1, a 42-year-old was submitted to PB biopsy with PGS due to advanced maternal age and poor ovarian reserve. Five MII oocytes underwent first and second polar body biopsy and four cleavage embryos were cryopreserved. The PGS analysis resulted in two euploid embryos (next generation sequence). A frozen-thawed embryo transfer (FET) was performed after endometrial priming and a healthy baby was delivered after a cesarean section (37 weeks, female, 3390g, 47.5 cm). In case 2, a 40-year old patient with balanced translocation and poor ovarian response was submitted to PB biopsy. Two MII oocytes underwent first and second polar body biopsy and two embryos were cryopreserved in cleavage stage. The analysis resulted in one euploid embryo that was transferred after endometrial priming. A preterm healthy baby (34 weeks, female, 2100g, 40 cm) was delivered via cesarean section. In conclusion, although the blastocyst biopsy is the norm when performing PGS/PGD during IVF treatments, other alternatives (as PB biopsy) should be considered in some specific situations.

Transcriptome profiling of human pre-implantation development.

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Pu Zhang

Full Text Available BACKGROUND: Preimplantation development is a crucial step in early human development. However, the molecular basis of human preimplantation development is not well known. METHODOLOGY: By applying microarray on 397 human oocytes and embryos at six developmental stages, we studied the transcription dynamics during human preimplantation development. PRINCIPAL FINDINGS: We found that the preimplantation development consisted of two main transitions: from metaphase-II oocyte to 4-cell embryo where mainly the maternal genes were expressed, and from 8-cell embryo to blastocyst with down-regulation of the maternal genes and up-regulation of embryonic genes. Human preimplantation development proved relatively autonomous. Genes predominantly expressed in oocytes and embryos are well conserved during evolution. SIGNIFICANCE: Our database and findings provide fundamental resources for understanding

The effect of nicotine pre-exposure on demand for cocaine and sucrose in male rats.

Science.gov (United States)

Schwartz, Lindsay P; Kearns, David N; Silberberg, Alan

2018-06-01

The aim of the present study was to determine how nicotine pre-exposure affects the elasticity of demand for intravenous cocaine and for sucrose pellets in adult male rats. In Experiment 1, demand for cocaine was assessed in rats that had nicotine in their drinking water. Nicotine pre-exposure significantly decreased rats' willingness to defend cocaine consumption as the price (measured as the number of responses per cocaine infusion) increased compared with a control group with no nicotine pre-exposure. That is, nicotine increased the elasticity of demand for cocaine infusions. Experiment 2 repeated the first experiment, but with rats working for sucrose pellets instead of cocaine. Nicotine pre-exposure had no effect on the elasticity of demand for sucrose. This pattern of results suggests that nicotine pre-exposure can reduce the reinforcing effects of cocaine, but not sucrose, in adult male rats.

The mutual influence of krypton implantation and pre-existing stress states in polycrystalline alpha titanium

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Nsengiyumva, S. [Department of Physics, University of Cape Town, Rondebosch 7701 (South Africa); Department of Physics, Kigali Institute of Education, P.O. Box 5039 Kigali (Rwanda)], E-mail: schadrack.nsengiyumva@uct.ac.za; Ntsoane, T.P. [Nuclear Energy Corporation of South Africa (NECSA), P.O. Box 582 (South Africa); Raji, A.T. [Department of Physics, University of Cape Town, Rondebosch 7701 (South Africa); Topic, M. [iThemba LABS, Somerset West 7129 (South Africa); Kellermann, G. [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas (Brazil); Riviere, J.P. [Laboratoire de Physique des Materiaux UMR6630-CNRS, 86960 (France); Britton, D.T.; Haerting, M. [Department of Physics, University of Cape Town, Rondebosch 7701 (South Africa)

2009-08-15

The stress profile in polycrystalline titanium implanted with krypton ions at different fluences has been determined using synchrotron radiation diffraction. For each fluence, the krypton profile has been measured using Rutherford backscattering geometry. The results were compared to model calculations obtained from the SRIM 2008 computer code. A strong stress relaxation was found for high fluence implantation, whereas for low fluence implantation an additional source of tensile stress was introduced in the near surface region. The projected range of the implanted krypton was significantly reduced compared to the expected range. A possible cause of this discrepancy is the drift of implanted ions under the influence of the pre-existing stress gradient.

The mutual influence of krypton implantation and pre-existing stress states in polycrystalline alpha titanium

International Nuclear Information System (INIS)

Nsengiyumva, S.; Ntsoane, T.P.; Raji, A.T.; Topic, M.; Kellermann, G.; Riviere, J.P.; Britton, D.T.; Haerting, M.

2009-01-01

The stress profile in polycrystalline titanium implanted with krypton ions at different fluences has been determined using synchrotron radiation diffraction. For each fluence, the krypton profile has been measured using Rutherford backscattering geometry. The results were compared to model calculations obtained from the SRIM 2008 computer code. A strong stress relaxation was found for high fluence implantation, whereas for low fluence implantation an additional source of tensile stress was introduced in the near surface region. The projected range of the implanted krypton was significantly reduced compared to the expected range. A possible cause of this discrepancy is the drift of implanted ions under the influence of the pre-existing stress gradient.

[Technical feasibility of the implantation of a monorail stent system into the renal arteries without pre-dilatation].

Science.gov (United States)

Neumann, C; Gschwendtner, M; Karnel, F; Mair, J; Dorffner, G; Dorffner, R

2005-01-01

To evaluate the technical feasibility of the implantation of the monorail RX Herculink system into the renal arteries without pre-dilatation. Forty-two patients (mean age 71 years) from four centers with a total of 44 renal artery stenoses underwent implantation of the RX Herculink stent. The mean grade of the stenosis was 83.8 %, the mean length 7.5 mm. The stenoses were ostial in 38 cases and in immediate proximity to the ostium in 6 cases. The mean follow-up-period was 57 weeks (24 - 176 weeks). In 42 cases, the implantation was successful without pre-dilatation. In 2 cases, pre-dilatation was carried out. In none of the cases, detachment of the stent from the balloon was observed. In one stenosis with a length of 17 mm, implantation of two stents was performed. In 9 cases, post-dilatation with a larger balloon or higher balloon pressure was necessary. Residual stenoses exceeding 30 % were not observed. Two patients developed local bleeding at the puncture site. During the follow-up, restenoses were observed in 5 stents after 26 to 126 weeks, which necessitated a second intervention in 3 cases (PTA in 2 cases, re-stenting in 1 case). The primary patency rate after 6 and 12 months was 0.92 +/- 0.056 according to Kaplan-Meier, the secondary patency rate after 6 and 12 months was 1.0 +/- 0.0. Implantation of the RX Herculink stent system into the renal arteries without pre-dilatation is technically feasible and safe. Even without pre-dilatation, the stent-system can be advanced through the stenosis without detachment. The complication rate is low. Our clinical results are comparable to previous studies.

A Randomized Trial to Evaluate the Effect of Local Endometrial Injury on the Clinical Pregnancy Rate of Frozen Embryo Transfer Cycles in Patients With Repeated Implantation Failure

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Ensieh Shahrokh-Tehraninejad

2016-12-01

Full Text Available Objective: Repeated implantation failure (RIF is a condition in which the embryos implantation decreases in the endometrium. So, our aim was to evaluate the effect of local endometrial injury on embryo transfer results.Materials and methods: In this simple randomized clinical trial (RCT, a total of 120 patients were selected. The participants were less than 40 years old, and they are in their minimum two cycles of vitro fertilization (IVF. Patients were divided randomly into two groups of LEI (Local endometrial injury and a control group (n = 60 in each group. The first group had four small endometrial injuries from anterior, posterior, and lateral uterus walls which were obtained from people who were in 21th day of their previous IVF cycle. The second group was the patients who have not received any intervention.Results: The experimental and control patients were matched in the following factors. Regarding the clinical pregnancy rate, there was no significant difference noted between the experimental and the control group.Conclusion: Local endometrial injury in a preceding cycle does not increase the clinical pregnancy rate in the subsequent FET cycle of patients with repeated implantation failure.

Pronuclear scoring as a predictor of embryo quality in in vitro fertilization program.

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Piotr Jedrzejczak

2008-04-01

Full Text Available Many strategies have been proposed for the selection of viable embryos for transfer in human assisted reproduction. These have included morphological scoring criteria for 20, 28, 44 and 68 h after insemination. The embryo selection is based on morphology, degree of fragmentation and development to the 8-cell. All have shown some correlation with implantation. However, the overall success of these methods is still limited, with over 50% of all transferred embryos failing to implant. Pronuclear zygote morphology has gained much attention recently due to its positive value in predicting implantation and pregnancy. This prospective study involved 178 conventional IVF patients only. The key aspects of pronuclear scoring and namely the presence of a cytoplasmic halo were related to day 3 of development and morphology in a retrospective study. The Z-score and the presence/absence of a halo had significant effect on the rate of development on day 3 embryo. Low Z-score result in slow development and poor morphology. The absence of a halo also resulted in slow and poor development, low morphology, increased fragmentation.

Non-invasive assessment of culture media from goat cloned embryos associated with subjective morphology by gas chromatography - mass spectroscopy-based metabolomic analysis.

Science.gov (United States)

Zhang, Yan-Li; Zhang, Guo-Min; Jia, Ruo-Xin; Wan, Yong-Jie; Yang, Hua; Sun, Ling-Wei; Han, Le; Wang, Feng

2018-01-01

Pre-implantation embryo metabolism demonstrates distinctive characteristics associated with the development potential of embryos. We aim to determine if metabolic differences correlate with embryo morphology. In this study, gas chromatography - mass spectroscopy (GC-MS)-based metabolomics was used to assess the culture media of goat cloned embryos collected from high-quality (HQ) and low-quality (LQ) groups based on morphology. Expression levels of amino acid transport genes were further examined by quantitative real-time PCR. Results showed that the HQ group presented higher percentages of blastocysts compared with the LQ counterparts (P culture media of the HQ group showed lower levels of valin, lysine, glutamine, mannose and acetol, and higher levels of glucose, phytosphingosine and phosphate than those of the LQ group. Additionally, expression levels of amino acid transport genes SLC1A5 and SLC3A2 were significantly lower in the HQ group than the LQ group (P culture media. The biochemical profiles may help to select the most in vitro viable embryos. © 2017 Japanese Society of Animal Science.

Influence of the time interval between embryo catheter loading and discharging on the success of IVF.

Science.gov (United States)

Matorras, R; Mendoza, R; Expósito, A; Rodriguez-Escudero, F J

2004-09-01

To ascertain the influence of the duration of the 'interval loading-discharging embryos' (ILDE) on the results of embryo transfer. The population under study consisted of 450 consecutive fresh embryo transfers. ILDE was measured in all transfers. Pregnancy and implantation rates were analyzed. Conceptional cycles had a lower (mean +/- SD) ILDE than non conceptional cycles (53.5 +/- 43.6 s vs 63.7 +/- 49.3). When only easy transfers--defined as those not requiring cervical tenaculum--were considered, similar differences were observed. The following pregnancy rates were obtained according ILDE duration: 38.9% (ILDE ILDE 31-60), 31.6% (ILDE 61-120) and 19.1% (ILDE > 120) (P ILDE duration is a prognostic factor of pregnancy rate and of implantation rate in IVF. The longer the ILDE duration, the lower the pregnancy and implantation rates. The decrease in pregnancy and implantation rates is gradual until an ILDE of 120 s, and decreases sharply afterwards. It is recommended to speed up the embryo transfer process, wherever possible. ILDE > 120 s carries a poor prognosis and should, when possible, be avoided.

The Impact of Biopsy on Human Embryo Developmental Potential during Preimplantation Genetic Diagnosis

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Danilo Cimadomo

2016-01-01

Full Text Available Preimplantation Genetic Diagnosis and Screening (PGD/PGS for monogenic diseases and/or numerical/structural chromosomal abnormalities is a tool for embryo testing aimed at identifying nonaffected and/or euploid embryos in a cohort produced during an IVF cycle. A critical aspect of this technology is the potential detrimental effect that the biopsy itself can have upon the embryo. Different embryo biopsy strategies have been proposed. Cleavage stage blastomere biopsy still represents the most commonly used method in Europe nowadays, although this approach has been shown to have a negative impact on embryo viability and implantation potential. Polar body biopsy has been proposed as an alternative to embryo biopsy especially for aneuploidy testing. However, to date no sufficiently powered study has clarified the impact of this procedure on embryo reproductive competence. Blastocyst stage biopsy represents nowadays the safest approach not to impact embryo implantation potential. For this reason, as well as for the evidences of a higher consistency of the molecular analysis when performed on trophectoderm cells, blastocyst biopsy implementation is gradually increasing worldwide. The aim of this review is to present the evidences published to date on the impact of the biopsy at different stages of preimplantation development upon human embryos reproductive potential.

SU-E-T-12: A Comparative Dosimetric Study of Pre and Post Prostate Iodine-125 Permanent Seed Implants

Energy Technology Data Exchange (ETDEWEB)

Liu, X; Rahimian, J; Goy, B; Cosmatos, H; Qian, Y [Kaiser Permanente, Los Angeles, CA (United States)

2015-06-15

Purpose: Post-implant dosimetry has become the gold standard for prostate implant evaluation. The goal of this research is to compare the dosimetry between pre-plan and post-plan in permanent prostate seed implant brachytherapy. Methods: A retrospective study of 91 patients treated with Iodine-125 prostate seed implant between year 2012∼2014 were performed. All plans were created using a VariSeed 8.0 planning system. Pre-plan ultrasound images were acquired using 0.5 cm slice thickness. Post-plan CT images acquired about 1–4 weeks after implant, fused with the preplan ultrasound images. The prostate and urethra contours were generated using the fusion of ultrasound and CT images. Iodine-125 seed source activities varied between 0.382 to 0.414 mCi per seed. The loading patterns varied slightly between patients depending on the prostate size. Statistical analysis of pre and post plans for prostate and urethra volumes, V100%, V150% and D90, and urethra D10 were performed and reported. Results: The pre and post implant average prostate size was 36.90cc vs. 38.58cc; V100% was 98.33% vs. 96.89%; V150% was 47.09% vs. 56.95%; D90 was 116.35Gy vs. 116.12Gy, urethra volume was 1.72cc vs. 1.85cc, urethra D10% was 122.0% vs. 135.35%, respectively. There was no statistically significant difference between the pre and post-plan values for D90(p-value=0.43). However, there are significant differences between other parameters most likely due to post surgical edema; prostate size (p-value= 0.00015); V100% (p-value=3.7803E-07); V150% (p-value=1.49E-09); urethra volume (p-value= 2.77E-06); Urethra D10 (p-value=7.37E-11). Conclusion: The post-plan dosimetry using CT image set showed similar D90 dose coverage to the pre-plan using the ultrasound image dataset. The study showed that our prostate seed implants have consistently delivered adequate therapeutic dose to the prostate while sparing urethra. Future studies to correlate dose versus biochemical response using patients’ PSA

The Histamine H1 Receptor Participates in the Increased Dorsal Telencephalic Neurogenesis in Embryos from Diabetic Rats

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Karina H. Solís

2017-12-01

Full Text Available Increased neuron telencephalic differentiation during deep cortical layer formation has been reported in embryos from diabetic mice. Transitory histaminergic neurons within the mesencephalon/rhombencephalon are responsible for fetal histamine synthesis during development, fibers from this system arrives to the frontal and parietal cortex at embryo day (E 15. Histamine is a neurogenic factor for cortical neural stem cells in vitro through H1 receptor (H1R which is highly expressed during corticogenesis in rats and mice. Furthermore, in utero administration of an H1R antagonist, chlorpheniramine, decreases the neuron markers microtubuline associated protein 2 (MAP2 and forkhead box protein 2. Interestingly, in the diabetic mouse model of diabetes induced with streptozotocin, an increase in fetal neurogenesis in terms of MAP2 expression in the telencephalon is reported at E11.5. Because of the reported effects on cortical neuron differentiation of maternal diabetes in one hand and of histamine in the other, here the participation of histamine and H1R on the increased dorsal telencephalic neurogenesis was explored. First, the increased neurogenesis in the dorsal telencephalon at E14 in diabetic rats was corroborated by immunohistochemistry and Western blot. Then, changes during corticogenesis in the level of histamine was analyzed by ELISA and in H1R expression by qRT-PCR and Western blot and, finally, we tested H1R participation in the increased dorsal telencephalic neurogenesis by the systemic administration of chlorpheniramine. Our results showed a significant increase of histamine at E14 and in the expression of the receptor at E12. The administration of chlorpheniramine to diabetic rats at E12 prevented the increased expression of βIII-tubulin and MAP2 mRNAs (neuron markers and partially reverted the increased level of MAP2 protein at E14, concluding that H1R have an important role in the increased neurogenesis within the dorsal telencephalon

Analysis of embryo morphokinetics, multinucleation and cleavage anomalies using continuous time-lapse monitoring in blastocyst transfer cycles.

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Desai, Nina; Ploskonka, Stephanie; Goodman, Linnea R; Austin, Cynthia; Goldberg, Jeffrey; Falcone, Tommaso

2014-06-20

Time-lapse imaging combined with embryo morphokinetics may offer a non-invasive means for improving embryo selection. Data from clinics worldwide are necessary to compare and ultimately develop embryo classifications models using kinetic data. The primary objective of this study was to determine if there were kinetic differences between embryos with limited potential and those more often associated with in vitro blastocyst formation and/or implantation. We also wanted to compare putative kinetic markers for embryo selection as proposed by other laboratories to what we were observing in our own laboratory setting. Kinetic data and cycle outcomes were retrospectively analyzed in patients age 39 and younger with 7 or more zygotes cultured in the Embryoscope. Timing of specific events from the point of insemination were determined using time-lapse (TL) imaging. The following kinetic markers were assessed: time to syngamy (tPNf), t2, time to two cells (c), 3c (t3), 4c ( t4), 5c (t5), 8c (t8), morula (tMor), start of blastulation (tSB); tBL, blastocyst (tBL); expanded blastocyst (tEBL). Durations of the second (cc2) and third (cc3) cell cycles, the t5-t2 interval as well as time to complete synchronous divisions s1, s2 and s3 were calculated. Incidence and impact on development of nuclear and cleavage anomalies were also assessed. A total of 648 embryos transferred on day 5 were analyzed. The clinical pregnancy and implantation rate were 72% and 50%, respectively. Morphokinetic data showed that tPNf, t2,t4, t8, s1, s2,s3 and cc2 were significantly different in embryos forming blastocysts (ET or frozen) versus those with limited potential either failing to blastulate or else forming poor quality blastocysts ,ultimately discarded. Comparison of embryo kinetics in cycles with all embryos implanting (KID+) versus no implantation (KID-) suggested that markers of embryo competence to implant may be different from ability to form a blastocyst. The incidence of multinucleation

Pulse of inflammatory proteins in the pregnant uterus of European polecats (Mustela putorius) leading to the time of implantation.

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Lindeberg, Heli; Burchmore, Richard J S; Kennedy, Malcolm W

2017-03-01

Uterine secretory proteins protect the uterus and conceptuses against infection, facilitate implantation, control cellular damage resulting from implantation, and supply pre-implantation embryos with nutrients. Unlike in humans, the early conceptus of the European polecat ( Mustela putorius ; ferret) grows and develops free in the uterus until implanting at about 12 days after mating. We found that the proteins appearing in polecat uteri changed dramatically with time leading to implantation. Several of these proteins have also been found in pregnant uteri of other eutherian mammals. However, we found a combination of two increasingly abundant proteins that have not been recorded before in pre-placentation uteri. First, the broad-spectrum proteinase inhibitor α 2 -macroglobulin rose to dominate the protein profile by the time of implantation. Its functions may be to limit damage caused by the release of proteinases during implantation or infection, and to control other processes around sites of implantation. Second, lipocalin-1 (also known as tear lipocalin) also increased substantially in concentration. This protein has not previously been recorded as a uterine secretion in pregnancy in any species. If polecat lipocalin-1 has similar biological properties to that of humans, then it may have a combined function in antimicrobial protection and transporting or scavenging lipids. The changes in the uterine secretory protein repertoire of European polecats is therefore unusual, and may be representative of pre-placentation supportive uterine secretions in mustelids (otters, weasels, badgers, mink, wolverines) in general.

Technical feasibility of the implantation of a monorail stent system into the renal arteries without pre-dilatation

International Nuclear Information System (INIS)

Neumann, C.; Dorffner, R.; Gschwendtner, M.; Karnel, F.; Mair, J.; Dorffner, G.

2005-01-01

Purpose: to evaluate the technical feasibility of the implantation of the monorail RX Herculink trademark system into the renal arteries without pre-dilatation. Materials and methods: forty-two patients (mean age 71 years) from four centers with a total of 44 renal artery stenoses underwent implantation of the RX Herculink trademark stent. The mean grade of the stenosis was 83.8%, the mean length 7.5 mm. The stenoses were ostial in 38 cases and in immediate proximity to the ostium in 6 cases. The mean follow-up-period was 57 weeks (24 - 176 weeks). Results: in 42 cases, the implantation was successful without pre-dilatation. In 2 cases, pre-dilatation was carried out. In none of the cases, detachment of the stent from the balloon was observed. In one stenosis with a length of 17 mm, implantation of two stents was performed. In 9 cases, post-dilatation with a larger balloon or higher balloon pressure was necessary. Residual stenoses exceeding 30% were not observed. Two patients developed local bleeding at the puncture site. During the follow-up, restenoses were observed in 5 stents after 26 to 126 weeks, which necessitated a second intervention in 3 cases (PTA in 2 cases, re-stenting in 1 case). The primary patency rate after 6 and 12 months was 0.92 ± 0.056 according to Kaplan-Meier, the secondary patency rate after 6 and 12 months was 1.0 ± 0.0. Conclusion: implantation of the RX Herculink trademark stent system into the renal arteries without pre-dilatation is technically feasible and safe. Even without pre-dilatation, the stent-system can be advanced through the stenosis without detachment. The complication rate is low. Our clinical results are comparable to previous studies. (orig.)

Influence of culture medium composition on relative mRNA abundances in domestic cat embryos.

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Hribal, R; Jewgenow, K; Braun, B C; Comizzoli, P

2013-04-01

Different culture conditions have been used to produce domestic cat embryos. As part of the in vitro procedures, the medium composition significantly affects the quality of the embryo development also. Quality assessments based on cleavage kinetics and blastomere symmetry are useful, but embryos also can differ in their relative gene expression patterns despite similar morphological characteristics. The aim of this study was to compare cat embryos produced with two different in vitro culture systems routinely used in two different laboratories [Smithsonian Conservation Biology Institute, Washington D.C., USA (SCBI) and Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany (IZW)]. Specifically, relative mRNA expression patterns of critical genes for pre-implantation embryo development were assessed in both conditions. Embryos were produced in parallel in both culture systems by IVF using frozen-thawed ejaculated semen in the United States and fresh epididymal sperm in Germany. Success of embryo development in vitro was recorded as well as relative mRNA abundances [DNA methyltransferases 1 and 3A (DNMT1, DNMT3A), gap junction protein alpha 1 (GJA1), octamer-binding transcription factor 4 [OCT4], insulin-like growth factors 1 and 2 receptors (IGF1R, IGF2R), beta-actin (ACTB)] in pools of days 4-5 morulae by semi-quantitative RT-PCR assay. Percentages of cleaved embryos were similar (p > 0.05) between both culture systems, regardless of the location. OCT4 mRNA abundance was higher (p culture system compared with those from the IZW system when epididymal sperm was used for IVF. No clear correlation between the expression pattern and the culture system could be found for all other genes. It is suggested that OCT4 expression might be affected by the media composition in some conditions and can be the indicator of a better embryo quality. © 2012 Blackwell Verlag GmbH.

Cryopreservation of peach palm zygotic embryos.

Science.gov (United States)

Steinmacher, Douglas A; Saldanha, Cleber W; Clement, Charles R; Guerra, Miguel P

2007-01-01

Cryopreservation is a safe and cost-effective option for long-term germplasm conservation of non-orthodox seed species, such as peach palm (Bactris gasipaes). The objective of the present study was to establish a cryopreservation protocol for peach palm zygotic embryos based on the encapsulation-dehydration technique. After excision, zygotic embryos were encapsulated with 3 percent sodium alginate plus 2 M glycerol and 0.4 M sucrose, and pre-treated or not with 1 M sucrose during 24 h, followed by air-drying. Fresh weight water contents of beads decreased from 83 percent and 87 percent to 18 percent and 20 percent for pre-treated or non-pretreated beads, respectively, after 4 h of dehydration. Sucrose pre-treatment at 1 M caused lower zygotic embryo germination and plantlet height in contrast to non-treated beads. All the variables were statistically influenced by dehydration time. Optimal conditions for recovery of cryopreserved zygotic embryos include encapsulation and dehydration for 4 h in a forced air cabinet to 20 percent water content, followed by rapid freezing in liquid nitrogen (-196 degree C) and rapid thawing at 45 degree C. In these conditions 29 percent of the zygotic embryos germinated in vitro. However, plantlets obtained from dehydrated zygotic embryos had stunted haustoria and lower heights. Histological analysis showed that haustorium cells were large, vacuolated, with few protein bodies. In contrast, small cells with high nucleus:cytoplasm ratio formed the shoot apical meristem of the embryos, which were the cell types with favorable characteristics for survival after exposure to liquid nitrogen. Plantlets were successfully acclimatized and showed 41+/-9 percent and 88+/-4 percent survival levels after 12 weeks of acclimatization from cryopreserved and non-cryopreserved treatments, respectively.

The lining of the gut in the developing rat embryo. Its relation to the hypoblast (primary endoderm) and the notochord

NARCIS (Netherlands)

Lamers, W. H.; Spliet, W. G.; Langemeyer, R. A.

1987-01-01

A light microscopical study of the morphogenesis of the gut in the rat embryo was undertaken to provide a careful map of temporal changes in the topographical relations of the (definitive) endoderm, the notochord and the hypoblast (primary endoderm). The borderline between the (definitive) endoderm

Gentamicin coating of plasma chemical oxidized titanium alloy prevents implant-related osteomyelitis in rats.

Science.gov (United States)

Diefenbeck, M; Schrader, C; Gras, F; Mückley, T; Schmidt, J; Zankovych, S; Bossert, J; Jandt, K D; Völpel, A; Sigusch, B W; Schubert, H; Bischoff, S; Pfister, W; Edel, B; Faucon, M; Finger, U

2016-09-01

Implant related infection is one of the most feared and devastating complication associated with the use of orthopaedic implant devices. Development of anti-infective surfaces is the main strategy to prevent implant contamination, biofilm formation and implant related osteomyelitis. A second concern in orthopaedics is insufficient osseointegration of uncemented implant devices. Recently, we reported on a macroporous titanium-oxide surface (bioactive TiOB) which increases osseointegration and implant fixation. To combine enhanced osseointegration and antibacterial function, the TiOB surfaces were, in addition, modified with a gentamicin coating. A rat osteomyelitis model with bilateral placement of titanium alloy implants was employed to analyse the prophylactic effect of gentamicin-sodiumdodecylsulfate (SDS) and gentamicin-tannic acid coatings in vivo. 20 rats were randomly assigned to four groups: (A) titanium alloy; PBS inoculum (negative control), (B) titanium alloy, Staphylococcus aureus inoculum (positive control), (C) bioactive TiOB with gentamicin-SDS and (D) bioactive TiOB plus gentamicin-tannic acid coating. Contamination of implants, bacterial load of bone powder and radiographic as well as histological signs of implant-related osteomyelitis were evaluated after four weeks. Gentamicin-SDS coating prevented implant contamination in 10 of 10 tibiae and gentamicin-tannic acid coating in 9 of 10 tibiae (infection prophylaxis rate 100% and 90% of cases, respectively). In Group (D) one implant showed colonisation of bacteria (swab of entry point and roll-out test positive for S. aureus). The interobserver reliability showed no difference in the histologic and radiographic osteomyelitis scores. In both gentamicin coated groups, a significant reduction of the histological osteomyelitis score (geometric mean values: C = 0.111 ± 0.023; D = 0.056 ± 0.006) compared to the positive control group (B: 0.244 ± 0.015; p < 0.05) was observed. The

Study of embryonic ploidy: a probable embryo model

Energy Technology Data Exchange (ETDEWEB)

Kundt, Miriam S; Cabrini, Romulo L [Comision Nacional de Energia Atomica, Buenos Aires (Argentina). Dept. de Radiobiologia

2001-07-01

The second polar body (PB) studies in preimplantation mouse embryos were carried out to evaluate the possibility as reference cell to analyze ploidy. For that purpose embryos in a one cell stage [obtained by crossing hybrid females (CBAxC57BL) to NIH males] were cultured in vitro during 72 hs, individually fixed at morula stage and stained with Feulgen. The DNA content of 263 individual nucleus was evaluated cytophotometrically corresponding to 22 compact morulas of normal development. As haploid PB is present in all pre implanted stage, only embryos with one haploid nuclei were considered as normal. In 95.5% (n = 21) of the embryos the PB was present. DNA measurement of 21 PB was 1n {+-} 0.1. By the height sensibility of PB ploidy, the abnormalities were detected by the criterion of >4.1 n and <1.9 n. The results showed that one embryo was completely haploid (1n). The rest of the embryos (n = 20) 222 blastomeres and 20 PB were analyzed. The DNA measurement showed that 92,7% of the blastomeres (n = 206) are between 2 n and 4 n and 7.3% showed ploidy anomalies, regarding the value n of their PB. The period of the cellular cycle was studied in the normal cell ploidy. This study showed that 16.5% of the blastomeres (n = 34) were in the period G1, 70.39% (n =34) in the period S and 13.2% in the period G2 (n = 27). It is concluded that the PB study showed that it has properties as an excellent indicator of internal ploidia: it is present from the moment of the conception, easily recognizable in the perivitelin space in the embryo of one-two cells, remains in interface during the preimplantation development, it is haploid and digitalized pixel by pixel PB study showed the homogeneity of this type of cell, giving a reliable value of ploidy. The properties of the PB and the results showed that the PB could be an excellent indicator for embryonic ploidy studies on genotoxicity, maintaining its original ploidia during the preimplantation development while the blastomeres are

Biodegradable Magnesium (Mg) Implantation Does Not Impose Related Metabolic Disorders in Rats with Chronic Renal Failure

Science.gov (United States)

Wang, Jiali; Xu, Jiankun; Liu, Waiching; Li, Yangde; Qin, Ling

2016-05-01

Mg and its alloys have been considered as one of the most promising biodegradable medical devices, but it was still unclear whether hypermagnesemia involved health risks would occur in persons with kidney disease due to their deteriorated kidney function for Mg ions excretion from their body. In this study, we established a chronic renal failure (CRF) model in rats induced by adenine administration prior to Mg implantation, aiming to predict if CRF patients are suitable for the use of Mg implants. The results showed that Mg levels in serum, urine, feces and internal organs had no significant changes after Mg implantation for both normal and CRF rats. Biochemical indices detection and histopathological analysis in kidney, liver and heart tissue confirmed that Mg implants did not induce any extra damage in animals even with renal failure. Our study indicates that Mg based orthopaedic medical device may be considered for use in CRF patients without biosafety concerns.

Effect of Fenugreek seed Extract (Trigonella Foenum-graecum on testicular tissue in the embryos of Streptozotocin Induced Diabetic Rats

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M beyzaei

2015-12-01

Full Text Available Background and aim: Diabetes mellitus is associated with some of the metabolic dysfunctions represented with chronic hyperglycemia.  This disease can disrupt the function of testicular tissue and decline male sexual ability. Some of the medicinal herbs such as fenugreeks have protective effects on tissues via hypoglycemic and anti-oxidative properties. In the present paper,  the effects of fenugreek seed extract was evaluated on testicular tissue of 20 day-old embryos from diabetic rats. Methods: In the present experimental study, sixty normal female rats were divided into three normal groups: non-diabetic control, glibenclamide and fenugreek groups and three diabetic groups: diabetic control, glibenclamide treatment and fenugreek treatment groups. Single injection of streptozotocin was used for induction of diabetes in these female rats. After detection of pregnancy, 1000 mg/kg fenugreek seed extract was fed to non-diabetic and diabetic fenugreek groups and 5 mg/kg glibenclamide was fed to non-diabetic and diabetic glibenclamide groups. Non-diabetic and diabetic control group was fed with distilled water as the same volume as the fenugreek extract. After 20 days, their embryos were pulled out and fixed at 10% formalin. After tissue processing, five micron sections were stained with Hematoxylin- eosin and evaluated for morphometric changes of testicular tissue. Data were evaluated with One-Way ANOVA test and Duncan post-hoc test. Results: The mean diameter of seminiferous tubules and testis capsule thickness indicated no significant differences between fenugreek treatment and diabetic control groups (P> 0.05. Mean body weight of male embryos was significantly lower in fenugreek treatment group in comparison with the diabetic control group (P&le 0.05. The leydig, sertoli and spermatogonial cells number was significantly higher in fenugreek treatment group in compression with diabetic control group                      (P

ENU mutagenesis reveals that Notchless homolog 1 (Drosophila affects Cdkn1a and several members of the Wnt pathway during murine pre-implantation development

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Lossie Amy C

2012-12-01

Full Text Available Abstract Background Our interests lie in determining the genes and genetic pathways that are important for establishing and maintaining maternal-fetal interactions during pregnancy. Mutation analysis targeted to a 34 Mb domain flanked by Trp53 and Wnt3 demonstrates that this region of mouse chromosome 11 contains a large number of essential genes. Two mutant alleles (l11Jus1 and l11Jus4, which fall into the same complementation group, survive through implantation but fail prior to gastrulation. Results Through a positional cloning strategy, we discovered that these homozygous mutant alleles contain non-conservative missense mutations in the Notchless homolog 1 (Drosophila (Nle1 gene. NLE1 is a member of the large WD40-repeat protein family, and is thought to signal via the canonical NOTCH pathway in vertebrates. However, the phenotype of the Nle1 mutant mice is much more severe than single Notch receptor mutations or even in animals in which NOTCH signaling is blocked. To test the hypothesis that NLE1 functions in multiple signaling pathways during pre-implantation development, we examined expression of multiple Notch downstream target genes, as well as select members of the Wnt pathway in wild-type and mutant embryos. We did not detect altered expression of any primary members of the Notch pathway or in Notch downstream target genes. However, our data reveal that Cdkn1a, a NOTCH target, was upregulated in Nle1 mutants, while several members of the Wnt pathway are downregulated. In addition, we found that Nle1 mutant embryos undergo caspase-mediated apoptosis as hatched blastocysts, but not as morulae or blastocysts. Conclusions Taken together, these results uncover potential novel functions for NLE1 in the WNT and CDKN1A pathways during embryonic development in mammals.

Biocompatibility of titanium based implants treated with plasma immersion ion implantation

International Nuclear Information System (INIS)

Maendl, S.; Sader, R.; Thorwarth, G.; Krause, D.; Zeilhofer, H.-F.; Horch, H.H.; Rauschenbach, B.

2003-01-01

In this work, the biocompatibility of titanium before and after oxygen PIII is investigated using a rat animal model. Pure titanium (grade 2) and pre-anodized titanium were implanted with oxygen at elevated temperatures between 200 and 550 deg. C and subsequently analyzed for oxygen content and phase composition. No deterioration of the tensile strength and the yield strength was detected after the implantation. The mechanical stability of the osseointegration was determined with a pull-out test, where an increased shear strength was measured after PIII treatment. Only a slight improvement of the bone contact area, from an already excellent starting value, was observed for pure titanium. In contrast, a significant improvement was found for anodized titanium after PIII treatment. This astonishing difference can be explained with the surface topography and the phase composition of the anodized titanium samples

Recurrent implantation failure: definition and management.

Science.gov (United States)

Coughlan, C; Ledger, W; Wang, Q; Liu, Fenghua; Demirol, Aygul; Gurgan, Timur; Cutting, R; Ong, K; Sallam, H; Li, T C

2014-01-01

Recurrent implantation failure refers to failure to achieve a clinical pregnancy after transfer of at least four good-quality embryos in a minimum of three fresh or frozen cycles in a woman under the age of 40 years. The failure to implant may be a consequence of embryo or uterine factors. Thorough investigations should be carried out to ascertain whether there is any underlying cause of the condition. Ovarian function should be assessed by measurement of antral follicle count, FSH and anti-Mu¨llerian hormone. Increased sperm DNA fragmentation may be a contributory cause. Various uterine pathology including fibroids, endometrial polyps, congenital anomalies and intrauterine adhesions should be excluded by ultrasonography and hysteroscopy. Hydrosalpinges are a recognized cause of implantation failure and should be excluded by hysterosalpingogram; if necessary, laparoscopy should be performed to confirm or refute the diagnosis. Treatment offered should be evidence based, aimed at improving embryo quality or endometrial receptivity. Gamete donation or surrogacy may be necessary if there is no realistic chance of success with further IVF attempts. Copyright © 2013. Published by Elsevier Ltd.

A novel knee prosthesis model of implant-related osteo- myelitis in rats

DEFF Research Database (Denmark)

Søe, Niels H.; Jensen, Nina Vendel; Nürnberg, Birgit Meinecke

2012-01-01

There have been numerous reports of animal models of osteomyelitis. Very few of these have been prosthesis models that imitate human conditions. We have developed a new rat model of implant-related osteomyelitis that mimics human osteomyelitis, to investigate the pathology of infection after...

Steroidal regulation of Ihh and Gli1 expression in the rat uterus.

Science.gov (United States)

Kubota, Kaiyu; Yamauchi, Nobuhiko; Yamagami, Kazuki; Nishimura, Sho; Gobaru, Takafumi; Yamanaka, Ken-ichi; Wood, Chris; Soh, Tomoki; Takahashi, Masashi; Hattori, Masa-aki

2010-05-01

Ovarian steroid hormones, progesterone (P4), and estradiol (E2) strictly regulate the endometrial tissue remodeling required for successful embryo implantation. Indian hedgehog (Ihh) is up-regulated by P4 and critically mediates uterine receptivity in the mouse. However, the regulation of Ihh expression during the implantation period still remains unclear. The present study was conducted to elucidate the mechanism of the steroidal regulation in the expression of Ihh and Gli1, the mediator of the Ihh pathway. Ihh mRNA was expressed in the rat uterus on 3.5-5.5 days post-coitus (dpc), while Gli1 expression transiently increased at 3.5 dpc but decreased significantly on 5.5 dpc (P Ihh was induced by the implantation-induced E2 treatment in the primed rat uterus. In contrast, expression of Gli1 was significantly decreased by E2 treatment (P = 0.016). In the case of ICI182.780 (ICI) treatment, Ihh expression was eliminated by ICI, whilst Gli1 expression increased. These results suggest that Ihh expression is maintained at a high level until the initiation of implantation, while the expression of Gli1 is decreased just prior to the initiation of implantation depending on the E2 action. This observation aids in the understanding of the Ihh signaling pathway mediating uterine remodeling for implantation.

Study of the boron levels in serum after implantation of different ratios nano-hexagonal boron nitride–hydroxy apatite in rat femurs

Energy Technology Data Exchange (ETDEWEB)

Atila, Alptug, E-mail: alptugatila@yahoo.com [Department of Analytical Chemistry, Faculty of Pharmacy, Ataturk University, Erzurum 25240 (Turkey); Halici, Zekai; Cadirci, Elif [Department of Pharmacology, Faculty of Medicine, Ataturk University, Erzurum 25240 (Turkey); Karakus, Emre [Department of Pharmacology and Toxicology, School of Veterinary Medicine, Ataturk University, Erzurum 25240 (Turkey); Palabiyik, Saziye Sezin [Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Ataturk University, Erzurum 25240 (Turkey); Ay, Nuran [Department of Material Science and Engineering, Faculty of Engineering, Anadolu University, Eskisehir 26555 (Turkey); Bakan, Feray [Sabancı University Nanotechnology Research and Application Center (SUNUM), Istanbul 34956 (Turkey); Yilmaz, Sahin [Department of Genetics and Bioengineering, Faculty of Engineering and Architecture, Yeditepe University, Istanbul 34755 (Turkey)

2016-01-01

ABSTRACT: Boron and its derivatives are effective in bone recovery and osteointegration. However, increasing the boron levels in body liquids may cause toxicity. The aim of our study is to investigate serum boron levels using ICP-MS after implantation of different ratios of nano-hBN–HA composites in rat femurs. All rats were (n = 126) divided into five experimental groups (n = 24) and one healthy group (6 rats); healthy (Group1), femoral defect + %100HA (Group2), femoral defect + %2.5hBN + %97.5HA (Group3), femoral defect + %5hBN + %95HA (Group4), femoral defect + %10hBN + %90 HA (Group5), femoral defect + %100hBN (Group6). The femoral defect was created in the distal femur (3 mm drill-bit). Each implant group was divided into four different groups (n = 24) also 6 rats sacrificed for each groups in one week intervals during four weeks. In our results; at 1, 2, 3, and 4 weeks after implantation near bone tissue, serum levels of boron were evaluated using ICP-MS. We demonstrated that neither short-term nor long-term implantation of hBN–HA composite resulted in statistically increased serum boron levels in experimental groups compared to healthy group. In conclusion, this study investigated the implant material produced form hBN–HA for the first time. Our data suggest that hBN is a new promising target for biomaterial and implant bioengineers. - Highlights: • Nano-hBN–HA composites are new targets for biomaterial and implant bioengineers. • Serum boron levels were researched after implantation of nano-hBN–HA composites. • Implantation of hBN–HA composite did not result in increased serum boron levels. • The use of boron in composite form with HA did not change the stability of the implant.

External fixation of femoral defects in athymic rats: Applications for human stem cell implantation and bone regeneration

Directory of Open Access Journals (Sweden)

Terasa Foo

2013-01-01

Full Text Available An appropriate animal model is critical for the research of stem/progenitor cell therapy and tissue engineering for bone regeneration in vivo. This study reports the design of an external fixator and its application to critical-sized femoral defects in athymic rats. The external fixator consists of clamps and screws that are readily available from hardware stores as well as Kirschner wires. A total of 35 rats underwent application of the external fixator with creation of a 6-mm bone defect in one femur of each animal. This model had been used in several separate studies, including implantation of collagen gel, umbilical cord blood mesenchymal stem cells, endothelial progenitor cells, or bone morphogenetic protein-2. One rat developed fracture at the proximal pin site and two rats developed deep tissue infection. Pin loosening was found in nine rats, but it only led to the failure of external fixation in two animals. In 8 to 10 weeks, various degrees of bone growth in the femoral defects were observed in different study groups, from full repair of the bone defect with bone morphogenetic protein-2 implantation to fibrous nonunion with collagen gel implantation. The external fixator used in these studies provided sufficient mechanical stability to the bone defects and had a comparable complication rate in athymic rats as in immunocompetent rats. The external fixator does not interfere with the natural environment of a bone defect. This model is particularly valuable for investigation of osteogenesis of human stem/progenitor cells in vivo.

Impact of follicular G-CSF quantification on subsequent embryo transfer decisions: a proof of concept study.

Science.gov (United States)

Lédée, N; Gridelet, V; Ravet, S; Jouan, C; Gaspard, O; Wenders, F; Thonon, F; Hincourt, N; Dubois, M; Foidart, J M; Munaut, C; Perrier d'Hauterive, S

2013-02-01

Previous experiments have shown that granulocyte colony-stimulating factor (G-CSF), quantified in the follicular fluid (FF) of individual oocytes, correlates with the potential for an ongoing pregnancy of the corresponding fertilized oocytes among selected transferred embryos. Here we present a proof of concept study aimed at evaluating the impact of including FF G-CSF quantification in the embryo transfer decisions. FF G-CSF was quantified with the Luminex XMap technology in 523 individual FF samples corresponding to 116 fresh transferred embryos, 275 frozen embryos and 131 destroyed embryos from 78 patients undergoing ICSI. Follicular G-CSF was highly predictive of subsequent implantation. The receiving operator characteristics curve methodology showed its higher discriminatory power to predict ongoing pregnancy in multivariate logistic regression analysis for FF G-CSF compared with embryo morphology [0.77 (0.69-0.83), P Embryos were classified by their FF G-CSF concentration: Class I over 30 pg/ml (a highest positive predictive value for implantation), Class II from 30 to 18.4 pg/ml and Class III Embryos derived from Class I follicles had a significantly higher implantation rate (IR) than those from Class II and III follicles (36 versus 16.6 and 6%, P Embryos derived from Class I follicles with an optimal morphology reached an IR of 54%. Frozen-thawed embryos transfer derived from Class I follicles had an IR of 37% significantly higher than those from Class II and III follicles, respectively, of 8 and 5% (P embryos but also 10% of the destroyed embryos were derived from G-CSF Class I follicles. Non-optimal embryos appear to have been transferred in 28% (22/78) of the women, and their pregnancy rate was significantly lower than that of women who received at least one optimal embryo (18 versus 36%, P = 0.04). Monitoring FF G-CSF for the selection of embryos with a better potential for pregnancy might improve the effectiveness of IVF by reducing the time and cost

The outcome of assisted hatching and defragmentation in patients with poor quality embryos on IVF outcome

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Ahmet Zeki Işık

2013-09-01

Full Text Available Objective: To evaluate the effects of assisted hatching in combination with defragmentation applied to day 3 poor quality human embryos, on implantation and pregnancy rates. Materials and Methods: A retrospective analysis was performed in patients (n=168 of all ages with no good quality transferable embryos (Veeck classification embryos >grade 1 and/or >10% fragmentation. The first group was consisted of cycles in which mAHA was performed to all transferred grade 2 embryos. The second group included transfer cycles where all the embryos were highly fragmented (between 10-50% fragmentations, grade 3 and aAHA and microsurgical fragment removal were applied to all of them. Results: In first group positive βhCG was 33%, clinical pregnancy (fetal heart beat rate was 27% and implantation rate was 11,28%. These rates were 37, 1%; 28, 8% and %16,19 respectively in the aAHA plus defragmentation group. In cases over 35 years of age in aAHA plus defragmentation group acceptable implantation (14,46% and clinical pregnancy (31,58% rates were achieved. Conclusion: In patients with no good quality transferable embryos AHA combined with defragmentation can be utilised with acceptable success rates in laboratories where there are experienced personnel available for this procedure otherwise only AHA can be the best option.

Use of micro-CT-based finite element analysis to accurately quantify peri-implant bone strains: a validation in rat tibiae.

Science.gov (United States)

Torcasio, Antonia; Zhang, Xiaolei; Van Oosterwyck, Hans; Duyck, Joke; van Lenthe, G Harry

2012-05-01

Although research has been addressed at investigating the effect of specific loading regimes on bone response around the implant, a precise quantitative understanding of the local mechanical response close to the implant site is still lacking. This study was aimed at validating micro-CT-based finite element (μFE) models to assess tissue strains after implant placement in a rat tibia. Small implants were inserted at the medio-proximal site of 8 rat tibiae. The limbs were subjected to axial compression loading; strain close to the implant was measured by means of strain gauges. Specimen-specific μFE models were created and analyzed. For each specimen, 4 different models were created corresponding to different representations of the bone-implant interface: bone and implant were assumed fully osseointegrated (A); a low stiffness interface zone was assumed with thickness of 40 μm (B), 80 μm (C), and 160 μm (D). In all cases, measured and computational strains correlated highly (R (2) = 0.95, 0.92, 0.93, and 0.95 in A, B, C, and D, respectively). The averaged calculated strains were 1.69, 1.34, and 1.15 times higher than the measured strains for A, B, and C, respectively, and lower than the experimental strains for D (factor = 0.91). In conclusion, we demonstrated that specimen-specific FE analyses provide accurate estimates of peri-implant bone strains in the rat tibia loading model. Further investigations of the bone-implant interface are needed to quantify implant osseointegration.

Embryotoxic and pharmacologic potency ranking of six azoles in the rat whole embryo culture by morphological and transcriptomic analysis

International Nuclear Information System (INIS)

Dimopoulou, Myrto; Verhoef, Aart; Pennings, Jeroen L.A.; Ravenzwaay, Bennard van; Rietjens, Ivonne M.C.M.; Piersma, Aldert H.

2017-01-01

Differential gene expression analysis in the rat whole embryo culture (WEC) assay provides mechanistic insight into the embryotoxicity of test compounds. In our study, we hypothesized that comparative analysis of the transcriptomes of rat embryos exposed to six azoles (flusilazole, triadimefon, ketoconazole, miconazole, difenoconazole and prothioconazole) could lead to a better mechanism-based understanding of their embryotoxicity and pharmacological action. For evaluating embryotoxicity, we applied the total morphological scoring system (TMS) in embryos exposed for 48 h. The compounds tested showed embryotoxicity in a dose-response fashion. Functional analysis of differential gene expression after 4 h exposure at the ID 10 (effective dose for 10% decreased TMS), revealed the sterol biosynthesis pathway and embryonic development genes, dominated by genes in the retinoic acid (RA) pathway, albeit in a differential way. Flusilazole, ketoconazole and triadimefon were the most potent compounds affecting the RA pathway, while in terms of regulation of sterol function, difenoconazole and ketoconazole showed the most pronounced effects. Dose-dependent analysis of the effects of flusilazole revealed that the RA pathway related genes were already differentially expressed at low dose levels while the sterol pathway showed strong regulation at higher embryotoxic doses, suggesting that this pathway is less predictive for the observed embryotoxicity. A similar analysis at the 24-hour time point indicated an additional time-dependent difference in the aforementioned pathways regulated by flusilazole. In summary, the rat WEC assay in combination with transcriptomics could add a mechanistic insight into the embryotoxic potency ranking and pharmacological mode of action of the tested compounds. - Highlights: • Embryonic exposure to azoles revealed concentration-dependent malformations. • Transcriptomics could enhance the mechanistic knowledge of embryotoxicants. • Retinoic

Embryotoxic and pharmacologic potency ranking of six azoles in the rat whole embryo culture by morphological and transcriptomic analysis

Energy Technology Data Exchange (ETDEWEB)

Dimopoulou, Myrto, E-mail: myrto.dimopoulou@wur.nl [Division of Toxicology, Wageningen University (Netherlands); National Institute of Public Health and the Environment (RIVM), Bilthoven (Netherlands); Verhoef, Aart; Pennings, Jeroen L.A. [National Institute of Public Health and the Environment (RIVM), Bilthoven (Netherlands); Ravenzwaay, Bennard van [Division of Toxicology, Wageningen University (Netherlands); BASF SE, Experimental Toxicology and Ecology, Ludwigshafen (Germany); Rietjens, Ivonne M.C.M. [Division of Toxicology, Wageningen University (Netherlands); Piersma, Aldert H. [National Institute of Public Health and the Environment (RIVM), Bilthoven (Netherlands); Institute for Risk Assessment Sciences, Utrecht University, Utrecht (Netherlands)

2017-05-01

Differential gene expression analysis in the rat whole embryo culture (WEC) assay provides mechanistic insight into the embryotoxicity of test compounds. In our study, we hypothesized that comparative analysis of the transcriptomes of rat embryos exposed to six azoles (flusilazole, triadimefon, ketoconazole, miconazole, difenoconazole and prothioconazole) could lead to a better mechanism-based understanding of their embryotoxicity and pharmacological action. For evaluating embryotoxicity, we applied the total morphological scoring system (TMS) in embryos exposed for 48 h. The compounds tested showed embryotoxicity in a dose-response fashion. Functional analysis of differential gene expression after 4 h exposure at the ID{sub 10} (effective dose for 10% decreased TMS), revealed the sterol biosynthesis pathway and embryonic development genes, dominated by genes in the retinoic acid (RA) pathway, albeit in a differential way. Flusilazole, ketoconazole and triadimefon were the most potent compounds affecting the RA pathway, while in terms of regulation of sterol function, difenoconazole and ketoconazole showed the most pronounced effects. Dose-dependent analysis of the effects of flusilazole revealed that the RA pathway related genes were already differentially expressed at low dose levels while the sterol pathway showed strong regulation at higher embryotoxic doses, suggesting that this pathway is less predictive for the observed embryotoxicity. A similar analysis at the 24-hour time point indicated an additional time-dependent difference in the aforementioned pathways regulated by flusilazole. In summary, the rat WEC assay in combination with transcriptomics could add a mechanistic insight into the embryotoxic potency ranking and pharmacological mode of action of the tested compounds. - Highlights: • Embryonic exposure to azoles revealed concentration-dependent malformations. • Transcriptomics could enhance the mechanistic knowledge of embryotoxicants. �

Characteristics of electrode impedance and stimulation efficacy of a chronic cortical implant using novel annulus electrodes in rat motor cortex

Science.gov (United States)

Wang, Chun; Brunton, Emma; Haghgooie, Saman; Cassells, Kahli; Lowery, Arthur; Rajan, Ramesh

2013-08-01

Objective. Cortical neural prostheses with implanted electrode arrays have been used to restore compromised brain functions but concerns remain regarding their long-term stability and functional performance. Approach. Here we report changes in electrode impedance and stimulation thresholds for a custom-designed electrode array implanted in rat motor cortex for up to three months. Main Results. The array comprises four 2000 µm long electrodes with a large annular stimulating surface (7860-15700 µm2) displaced from the penetrating insulated tip. Compared to pre-implantation in vitro values there were three phases of impedance change: (1) an immediate large increase of impedance by an average of two-fold on implantation; (2) a period of continued impedance increase, albeit with considerable variability, which reached a peak at approximately four weeks post-implantation and remained high over the next two weeks; (3) finally, a period of 5-6 weeks when impedance stabilized at levels close to those seen immediately post-implantation. Impedance could often be temporarily decreased by applying brief trains of current stimulation, used to evoke motor output. The stimulation threshold to induce observable motor behaviour was generally between 75-100 µA, with charge density varying from 48-128 µC cm-2, consistent with the lower current density generated by electrodes with larger stimulating surface area. No systematic change in thresholds occurred over time, suggesting that device functionality was not compromised by the factors that caused changes in electrode impedance. Significance. The present results provide support for the use of annulus electrodes in future applications in cortical neural prostheses.

Human implantation: the last barrier in assisted reproduction technologies?

Science.gov (United States)

Edwards, Robert G

2006-12-01

Implantation processes are highly complex involving the actions of numerous hormones, immunoglobulins, cytokines and other factors in the endometrium. They are also essential matters for the success of assisted reproduction. The nature of early embryonic development is of equal significance. It involves ovarian follicle growth, ovulation, fertilization and preimplantation growth. These processes are affected by imbalanced chromosomal constitutions or slow developmental periods. Post-implantation death is also a significant factor in cases of placental insufficiency or recurrent abortion. Clearly, many of these matters can significantly affect birth rates. This review is concerned primarily with the oocyte, the early embryo and its chromosomal anomalies, and the nature of factors involved in implantation. These are clearly among the most important features in determining successful embryonic and fetal growth. Successive sections cover the endocrine stimulation of follicle growth in mice and humans, growth of human embryos in vitro, their apposition and attachment to the uterus, factors involved in embryo attachment to uterine epithelium and later stages of implantation, and understanding the gene control of polarities and other aspects of preimplantation embryo differentiation. New aspects of knowledge include the use of human oocyte maturation in vitro as an approach to simpler forms of IVF, and new concepts in developmental genetics.

Rat primary embryo fibroblast cells suppress transformation by the E6 and E7 genes of human papillomavirus type 16 in somatic hybrid cells.

OpenAIRE

Miyasaka, M; Takami, Y; Inoue, H; Hakura, A

1991-01-01

The E6 and E7 genes of human papillomavirus type 16 (HPV-16) transform established lines of rat cells but not rat cells in primary culture irrespective of the expression of the two genes. The reason for this difference between the susceptibilities of cell lines and primary cells was examined by using hybrid cells obtained by somatic cell fusion of rat cell lines transformed by the E6 and E7 genes of HPV-16 and freshly isolated rat embryo fibroblast cells. In these hybrid cells, transformed ph...

Model of human recurrent respiratory papilloma on chicken embryo chorioallantoic membrane for tumor angiogenesis research.

Science.gov (United States)

Uloza, Virgilijus; Kuzminienė, Alina; Palubinskienė, Jolita; Balnytė, Ingrida; Ulozienė, Ingrida; Valančiūtė, Angelija

2017-07-01

We aimed to develop a chick embryo chorioallantoic membrane (CAM) model of recurrent respiratory papilloma (RPP) and to evaluate its morphological and morphometric characteristics, together with angiogenic features. Fresh RRP tissue samples obtained from 13 patients were implanted in 174 chick embryo CAMs. Morphological, morphometric, and angiogenic changes in the CAM and chorionic epithelium were evaluated up until 7 days after the implantation. Immunohistochemical analysis (34βE12, Ki-67, MMP-9, PCNA, and Sambucus nigra staining) was performed to detect cytokeratins and endothelial cells and to evaluate proliferative capacity of the RRP before and after implantation on the CAM. The implanted RRP tissue samples survived on CAM in 73% of cases while retaining their essential morphologic characteristics and proliferative capacity of the original tumor. Implants induced thickening of both the CAM (241-560%, p=0.001) and the chorionic epithelium (107-151%, p=0.001), while the number of blood vessels (37-85%, p=0.001) in the CAM increased. The results of the present study confirmed that chick embryo CAM is a relevant host for serving as a medium for RRP fresh tissue implantation. The CAM assay demonstrated the specific RRP tumor growth pattern after implantation and provided the first morphological and morphometric characterization of the RRP CAM model that opens new horizons in studying this disease.

Comparing rat and rabbit embryo-fetal developmental toxicity studies for 379 pharmaceuticals: On systemic dose and developmental effects (Critical Reviews in Toxicology)

Science.gov (United States)

A database of embryo-fetal developmental toxicity (EFDT) studies of 379 pharmaceutical compounds in rat and rabbit was analyzed for species differences based on toxicokinetic parameters of area under the curve (AUC) and maximum concentration (Cmax) at the developmental adverse ef...

Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein.

Science.gov (United States)

Ideta, Atsushi; Aoyagi, Yoshito; Tsuchiya, Kanami; Nakamura, Yuuki; Hayama, Kou; Shirasawa, Atsushi; Sakaguchi, Kenichiro; Tominaga, Naomi; Nishimiya, Yoshiyuki; Tsuda, Sakae

2015-01-01

Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24-48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP-embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.

The effect of therapeutic ultrasound on metallic implants: a study in rats.

Science.gov (United States)

Kocaoğlu, Barış; Cabukoglu, Cengiz; Ozeras, Nadire; Seyhan, Mustafa; Karahan, Mustafa; Yalcin, Selim

2011-11-01

To investigate whether therapeutic ultrasound (US) use over metallic implants has the potential for adverse effects as a result of greater temperature increases at the tissue-metal interface. A randomized controlled trial. A research laboratory. Sprague-Dawley rats (N=40; weight, 230-300g) were used and divided into 3 study groups. In group 1, both limbs of 10 rats were used for evaluation of temperature changes. Metal pins were placed into the femur of the left limb, and the right limbs were used as controls. A thermal sensor was placed into the medulla to record the elevation of tissue temperature during US application. In groups 2 and 3 with 15 rats in each, a midshaft femoral fracture was produced, and intramedullary fixation was performed with metal pins. Group 2 received US treatment for 5 minutes daily and continued for 27 days. Group 3 served as controls. The rats in groups 2 and 3 were killed on postoperative day 30. The specimens were evaluated by radiology, histopathology, and biomechanics. The presence of metal in bone did not cause an increased temperature rise. US application did not increase or decrease callus formation, and there was no tissue necrosis. The average removal torques of pins in groups did not show a significant difference. Internal fixation with metallic implants may not be a contraindication for therapeutic US treatment. Copyright © 2011 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

Insight into PreImplantation Factor (PIF* mechanism for embryo protection and development: target oxidative stress and protein misfolding (PDI and HSP through essential RIKP [corrected] binding site.

Directory of Open Access Journals (Sweden)

Eytan R Barnea

Full Text Available Endogenous PIF, upon which embryo development is dependent, is secreted only by viable mammalian embryos, and absent in non-viable ones. Synthetic PIF (sPIF administration promotes singly cultured embryos development and protects against their demise caused by embryo-toxic serum. To identify and characterize critical sPIF-embryo protein interactions novel biochemical and bio-analytical methods were specifically devised.FITC-PIF uptake/binding by cultured murine and equine embryos was examined and compared with scrambled FITC-PIF (control. Murine embryo (d10 lysates were fractionated by reversed-phase HPLC, fractions printed onto microarray slides and probed with Biotin-PIF, IDE and Kv1.3 antibodies, using fluorescence detection. sPIF-based affinity column was developed to extract and identify PIF-protein interactions from lysates using peptide mass spectrometry (LC/MS/MS. In silico evaluation examined binding of PIF to critical targets, using mutation analysis.PIF directly targets viable cultured embryos as compared with control peptide, which failed to bind. Multistep Biotin-PIF targets were confirmed by single-step PIF-affinity column based isolation. PIF binds protein disulfide isomerases a prolyl-4-hydroxylase β-subunit, (PDI, PDIA4, PDIA6-like containing the antioxidant thioredoxin domain. PIF also binds protective heat shock proteins (70&90, co-chaperone, BAG-3. Remarkably, PIF targets a common RIKP [corrected] site in PDI and HSP proteins. Further, single PIF amino acid mutation significantly reduced peptide-protein target bonding. PIF binds promiscuous tubulins, neuron backbones and ACTA-1,2 visceral proteins. Significant anti-IDE, while limited anti-Kv1.3b antibody-binding to Biotin-PIF positive lysates HPLC fractions were documented.Collectively, data identifies PIF shared targets on PDI and HSP in the embryo. Such are known to play a critical role in protecting against oxidative stress and protein misfolding. PIF-affinity-column is a

Influences of somatic donor cell sex on and embryo development following somatic cell nuclear transfer in pigs

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Jae-Gyu Yoo

2017-04-01

Full Text Available Objective The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture (143.8±10.5 to 159.2±14.8 was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT groups (31.4±8.3 to 33.4±11.1. After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p<0.05 between sexes. Conclusion The present study found that the sex difference of the nuclear donor does not affect the developmental rate of porcine SCNT embryos. Furthermore, postnatal survivability of the cloned piglets was not affected by the sex of the donor cell.

CT pre-operative planning of a new semi-implantable bone conduction hearing device

Energy Technology Data Exchange (ETDEWEB)

Law, Eric K.C.; Bhatia, Kunwar S.S. [Prince of Wales Hospital, The Chinese University of Hong Kong, Department of Imaging and Interventional Radiology, Hong Kong, SAR (China); Tsang, Willis S.S.; Tong, Michael C.F. [Prince of Wales Hospital, The Chinese University of Hong Kong, Department of Otorhinolaryngology, Head and Neck Surgery, Hong Kong, SAR (China); Shi, Lin [The Chinese University of Hong Kong, Department of Medicine and Therapeutics, Hong Kong, SAR (China); The Chinese University of Hong Kong, Chow Yuk Ho Technology Center for Innovative Medicine, Hong Kong, SAR (China)

2016-06-15

Accommodating a novel semi-implantable bone conduction hearing device within the temporal bone presents challenges for surgical planning. This study describes the utility of CT in pre-operative assessment of such an implant. Retrospective review of pre-operative CT, clinical and surgical records of 16 adults considered for device implantation. Radiological suitability was assessed on CT using 3D simulation software. Antero-posterior (AP) dimensions of the mastoid bone and minimum skull thickness were measured. CT planning results were correlated with operative records. Eight and five candidates were suitable for device placement in the transmastoid and retrosigmoid positions, respectively, and three were radiologically unsuitable. The mean AP diameter of the mastoid cavity was 14.6 mm for the transmastoid group and 4.6 mm for the retrosigmoid group (p < 0.05). Contracted mastoid and/or prior surgery were predisposing factors for unsuitability. Four transmastoid and five retrosigmoid positions required sigmoid sinus/dural depression and/or use of lifts due to insufficient bone capacity. A high proportion of patients being considered have contracted or operated mastoids, which reduces the feasibility of the transmastoid approach. This finding combined with the complex temporal bone geometry illustrates the importance of careful CT evaluation using 3D software for precise device simulation. (orig.)

Alveolar wound healing after implantation with a pool of commercially available bovine bone morphogenetic proteins (BMPs): a histometric study in rats.

Science.gov (United States)

Calixto, Romeu Felipe Elias; Teófilo, Juliana Mazzonetto; Brentegani, Luiz Guilherme; Lamano-Carvalho, Teresa Lúcia

2007-01-01

The capacity of a commercially available pool of bovine bone morphogenetic proteins (BMPs) to stimulate osteogenesis in the rat alveolar healing was investigated by histometric analysis. Male rats were anesthetized and had their upper incisor extracted. A pool of purified bovine BMPs adsorbed to microgranular resorbable hydroxyapatite was agglutinated with bovine collagen and saline before implantation into the alveolar socket. The implanted and control rats (n=30 per group) were sacrificed 1 to 9 weeks postoperatively, the hemi-maxillae were decalcified, processed for paraffin embedding and semi-serial longitudinal sections were obtained and stained with hematoxylin and eosin. The volume fraction of alveolar healing components was estimated by a differential point-counting method in histologic images. The results showed that in both, control and implanted rats, the alveolar healing followed the histologic pattern usually described in the literature. Quantitative data confirmed that the BMPs mixture did not stimulate new bone formation in the alveolar socket of implanted rats. These results suggest that the pool of BMPs adsorbed to hydroxyapatite and agglutinated with bovine collagen did not warrant incorporation of the osteoinductive proteins to a slow-absorption system that would allow a BMPs release rate compatible to that of new bone formation, and thus more adequate to osteoinduction.

Influence of the radiation (Co{sub 60}) in pre-implants rabbit embryos: effect on atypic mitotic index and embryo pole development; Influencia da radiacao ionizante (bomba de cobalto) em embrioes de coelha na fase de pre-implantacao: influencia no numero de mitoses atipicas e no grau de desenvolvimento do polo

Energy Technology Data Exchange (ETDEWEB)

Approbato, Mario S.; Oliveira Moura, Katia K.V. de; Souza Florencio, Rodopiano de; Garcia, Ricardo; Faria, Renato S.; Benedetti, Leonardo N.; Goulart, Flamarion B. [Goias Univ., Goiania, GO (Brazil). Faculdade de Medicina. Dept. de Ginecologia e Obstetricia

1995-08-01

We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), at three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses: five c Gy and ten c Gy. Six rabbits (36 blastocysts) were used as controls. the matching instant was the zero hour. Exactly six days after ({+-} 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. the embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryo parameters were studied: embryo pole development; percentage of abnormal mitotic figures. irradiation time was associated with lower scores of embryo pole development, but not with irradiation dose. There were no gross abnormalities of embryo pole. The abnormal mitotic cells was affected both by the time and dose of irradiation. (author). 12 refs., 4 figs.

Study of the boron levels in serum after implantation of different ratios nano-hexagonal boron nitride-hydroxy apatite in rat femurs.

Science.gov (United States)

Atila, Alptug; Halici, Zekai; Cadirci, Elif; Karakus, Emre; Palabiyik, Saziye Sezin; Ay, Nuran; Bakan, Feray; Yilmaz, Sahin

2016-01-01

Boron and its derivatives are effective in bone recovery and osteointegration. However, increasing the boron levels in body liquids may cause toxicity. The aim of our study is to investigate serum boron levels using ICP-MS after implantation of different ratios of nano-hBN-HA composites in rat femurs. All rats were (n=126) divided into five experimental groups (n=24) and one healthy group (6 rats); healthy (Group1), femoral defect + %100 HA (Group2), femoral defect + %2.5 hBN + %97.5 HA (Group3), femoral defect + %5 hBN + %95 HA (Group4), femoral defect + %10 hBN + %90 HA (Group5), femoral defect + %100 hBN (Group6). The femoral defect was created in the distal femur (3mm drill-bit). Each implant group was divided into four different groups (n=24) also 6 rats sacrificed for each groups in one week intervals during four weeks. In our results; at 1, 2, 3, and 4 weeks after implantation near bone tissue, serum levels of boron were evaluated using ICP-MS. We demonstrated that neither short-term nor long-term implantation of hBN-HA composite resulted in statistically increased serum boron levels in experimental groups compared to healthy group. In conclusion, this study investigated the implant material produced form hBN-HA for the first time. Our data suggest that hBN is a new promising target for biomaterial and implant bioengineers. Copyright © 2015 Elsevier B.V. All rights reserved.

Brain activation in response to visceral stimulation in rats with amygdala implants of corticosterone: an FMRI study.

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Anthony C Johnson

2010-01-01

Full Text Available Although visceral pain of gastrointestinal (GI origin is the major complaint in patients with irritable bowel syndrome (IBS it remains poorly understood. Brain imaging studies suggest a defect in brain-gut communication in IBS with a greater activation of central arousal circuits including the amygdala. Previously, we found that stereotaxic implantation of corticosterone (CORT onto the amygdala in rats induced anxiety and colonic hypersensitivity. In the present study we used functional magnetic resonance imaging (fMRI to identify specific brain sites activated in a rat model characterized by anxiety and colonic hypersensitivity.Anesthetized male rats received micropellets (30 microg each of either CORT or cholesterol (CHOL, to serve as a control, implanted stereotaxically on the dorsal margin of each amygdala. Seven days later, rats were anesthetized and placed in the fMRI magnet (7T. A series of isobaric colorectal balloon distensions (CRD - 90s 'off', 30s 'on', 8 replicates at two pressures (40 and 60 mmHg were performed in a standard block-design. Cross correlation statistical analysis was used to determine significant differences between distended and non-distended states in CORT and CHOL-treated animals. Analysis of the imaging data demonstrated greater overall brain activation in response to CRD in rats with CORT implants compared to CHOL controls. Additionally, CORT implants produced significant positive bilateral increases in MRI signal in response to CRD in specific nuclei known as integration sites important in anxiety and pain perception.These data indicate that chronic exposure of the amygdala to elevated levels of CORT enhances overall brain activation in response to CRD, and identified other specific brain regions activated in response to mechanical distension of the colon. These results demonstrate the feasibility of performing fMRI imaging in a rodent model that supports clinical observations in IBS patients with enhanced

Characterization and quantification of proteins secreted by single human embryos prior to implantation.

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Poli, Maurizio; Ori, Alessandro; Child, Tim; Jaroudi, Souraya; Spath, Katharina; Beck, Martin; Wells, Dagan

2015-11-01

The use of in vitro fertilization (IVF) has revolutionized the treatment of infertility and is now responsible for 1-5% of all births in industrialized countries. During IVF, it is typical for patients to generate multiple embryos. However, only a small proportion of them possess the genetic and metabolic requirements needed in order to produce a healthy pregnancy. The identification of the embryo with the greatest developmental capacity represents a major challenge for fertility clinics. Current methods for the assessment of embryo competence are proven inefficient, and the inadvertent transfer of non-viable embryos is the principal reason why most IVF treatments (approximately two-thirds) end in failure. In this study, we investigate how the application of proteomic measurements could improve success rates in clinical embryology. We describe a procedure that allows the identification and quantification of proteins of embryonic origin, present in attomole concentrations in the blastocoel, the enclosed fluid-filled cavity that forms within 5-day-old human embryos. By using targeted proteomics, we demonstrate the feasibility of quantifying multiple proteins in samples derived from single blastocoels and that such measurements correlate with aspects of embryo viability, such as chromosomal (ploidy) status. This study illustrates the potential of high-sensitivity proteomics to measure clinically relevant biomarkers in minute samples and, more specifically, suggests that key aspects of embryo competence could be measured using a proteomic-based strategy, with negligible risk of harm to the living embryo. Our work paves the way for the development of "next-generation" embryo competence assessment strategies, based on functional proteomics. © 2015 The Authors. Published under the terms of the CC BY 4.0 license.

Ultrastructural Modifications of Human Endometrium during the Window of Implantation

Directory of Open Access Journals (Sweden)

Maryam Kabir-Salmani

2008-01-01

Full Text Available The endometrium is a highly dynamic tissue empowered with the capacity to undergo cyclicdramatic changes in response to ovarian steroid hormones, ultimately aiming to create awindow of receptivity for blastocyst implantation. Intensive research has been performed tounderstand and establish morphological and molecular correlates of embryo implantation.However, it still remains a biological mystery particularly in the human, where ethical andmoral constraints prohibit in vivo testing and the establishment of an ideal in vitro modeling.Rodent models of embryo implantation are largely irrelevant because the process variessignificantly from that in humans. Even among primates, subtle differences exist amongspecies. For maternal preparation of implantation, the endometrial epithelium which issurprisingly hostile towards the embryo implantation, acquires functional status receptiveto blastocyst acceptance during a limited period of cycle days, termed as the ‘window ofimplantation (WOI. This review provides currently available information concernedprimarily with the various ultrastructural modifications of endometrium coordinated withinthe WOI that may signify endometrial receptivity. In the following sections, the dominantfeatures of endometrial differentiation during WOI, including transformations of luminalepithelium, endometrial glands, and stromal decidualization will be discussed from themorphological points of view.

Moving and fusion of the pancreatic buds in the rat embryos during the embryonic period (carnegie stages 13-17) by a three-dimensional computer-assisted reconstruction.

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Godlewski, Guilhem; Gaubert, Jacques; Cristol-Gaubert, Renée; Radi, Maïada; Baecker, Volker; Travo, Pierre; Prudhomme, Michel; Prat-Pradal, Dominique

2011-10-01

The purpose of the present study was to illustrate the modality of rotation of ventral and dorsal pancreatic buds by three-dimensional (3D) reconstructions in the rat embryos, during the Carnegie stages 13-17. Serial sections of thirty rat embryos stages 13-17, were observed. The embryos were fixed in Bouin's solution, dehydrated, and paraffin embedded. The sections, 7 μm thick, were cut in longitudinal or transverse planes and were stained alternately by hematoxylin-eosin or Heindenhain' azan. The images were digitalized by Canon Camera 350 EOS D. The 3D reconstruction was performed by computer using Cell Image Analyser software. The two pancreatic buds ventral and dorsal, were clearly identified at stage 13, in anterior and posterior position, respectively, in relation to the duodenum. In stage 15, the duodenum started its rotation of 90° clockwise. The ventral bud moved 90° from the midline to the right. In stage 16, the ventral pancreas continued its rotation until 180° in posterior position behind the duodenum. In stage 17, the two pancreatic buds were related closely to the ventral part of the portal vein. The two buds began to merge. The anterior face of the pancreas's head was arising from the dorsal pancreatic bud. The rest of the head including the omental tuberosity and the uncinate process emanated from the ventral pancreatic bud. The use of 3D reconstruction of the pancreas of rat embryos illustrates the modality of the two pancreatic buds rotation and fusion. This method explains the final position of the pancreas.

Importance of Maternal Diabetes on the Chronological Deregulation of the Intrauterine Development: An Experimental Study in Rat

Science.gov (United States)

Salazar García, Marcela; Reyes Maldonado, Elba; Revilla Monsalve, María Cristina; Villavicencio Guzmán, Laura; Reyes López, Alfonso; Sánchez-Gómez, Concepción

2015-01-01

We investigated whether maternal diabetes induced in rats using streptozotocin (STZ) on Day 5 of pregnancy affects the intrauterine developmental timeline. A total of 30 pregnant Sprague-Dawley diabetic rats (DRs) and 20 control rats (CRs) were used to obtain 21-day fetuses (F21) and newborn (NB) pups. Gestational age, weight, and body size were recorded as were the maxillofacial morphometry and morphohistological characteristics of the limbs. In DRs, pregnancy continued for ∼1.7 days, and delivery occurred 23 days postcoitus (DPC). In this group, the number of pups was lower, and 13% had maxillofacial defects. F21 in the DR group had lower weights and were smaller; moreover, the morphological characteristics of the maxillofacial structures, derived from the neural crest, were discordant with their chronological gestational age, resembling 18- to 19-day-old fetuses. These deficiencies were counterbalanced in NB pups. We conclude that hyperglycemia, which results from maternal diabetes and precedes embryo implantation, deregulates the intrauterine developmental timeline, restricts embryo-fetal growth, and primarily delays the remodeling and maturation of the structures derived from neural crest cells. PMID:25756053

Low versus high volume of culture medium during embryo transfer: a randomized clinical trial.

Science.gov (United States)

Sigalos, George Α; Michalopoulos, Yannis; Kastoras, Athanasios G; Triantafyllidou, Olga; Vlahos, Nikos F

2018-04-01

The aim of this prospective randomized control trial was to evaluate if the use of two different volumes (20-25 vs 40-45 μl) of media used for embryo transfer affects the clinical outcomes in fresh in vitro fertilization (IVF) cycles. In total, 236 patients were randomized in two groups, i.e., "low volume" group (n = 118) transferring the embryos with 20-25 μl of medium and "high volume" group (n = 118) transferring the embryos with 40-45 μl of medium. The clinical pregnancy, implantation, and ongoing pregnancy rates were compared between the two groups. No statistically significant differences were observed in clinical pregnancy (46.8 vs 54.3%, p = 0.27), implantation (23.7 vs 27.8%, p = 0.30), and ongoing pregnancy (33.3 vs 40.0%, p = 0.31) rates between low and high volume group, respectively. Higher volume of culture medium to load the embryo into the catheter during embryo transfer does not influence the clinical outcome in fresh IVF cycles. NCT03350646.

Insulin-like growth factor binding protein-3 affects osteogenic efficacy on dental implants in rat mandible

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Bhattarai, Govinda; Lee, Young-Hee [Department of Oral Biochemistry, Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju (Korea, Republic of); Lee, Min-Ho [Department of Dental Materials, Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju (Korea, Republic of); Park, Il-Song [Division of Advanced Materials Engineering, Research Center for Advanced Materials, Development and Institute of Biodegradable Materials, Chonbuk National University, Jeonju 561-756 (Korea, Republic of); Yi, Ho-Keun, E-mail: yihokn@chonbuk.ac.kr [Department of Oral Biochemistry, Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju (Korea, Republic of)

2015-10-01

Insulin like growth factor binding protein-3 (IGFBP-3) in bone cells and its utilization in dental implants have not been well studied. The aim of this study was to determine the osteogenic efficacy of chitosan gold nanoparticles (Ch-GNPs) conjugated with IGFBP-3 coated titanium (Ti) implants. Ch-GNPs were conjugated with IGFBP-3 plasmid DNA through a coacervation process. Conjugation was cast over Ti surfaces, and cells were seeded on coated surfaces. For in vitro analysis the expression of different proteins was analyzed by immunoblotting. For in vivo analysis, Ch-GNP/IGFBP-3 coated implants were installed in rat mandibles. Four weeks post-implantation, mandibles were examined by microcomputed tomography (μCT), immunohistochemistry, hematoxylin & eosin and tartrate resistance acid phosphatase staining. In vitro overexpressed Ch-GNP/IGFBP-3 coated Ti surfaces was associated with activation of extracellular signal related kinase (ERK), inhibition of the stress activated protein c-Jun N-terminal kinase (JNK) and enhanced bone morphogenetic protein (BMP)-2 and 7 compared to control. Further, in vivo, Ch-GNP/IGFBP-3 coated implants were associated with inhibition of implant induced osteoclastogenesis molecules, receptor activator of nuclear factor kappa-B ligand (RANKL) and enhanced expression of osteogenic molecules including BMP2/7 and osteopontin (OPN). The μCT analysis demonstrated that IGFBP-3 increased the volume of newly formed bone surrounding the implants compared to control (n = 5; p < 0.05). These results support the view that IGFBP-3 overexpression diminishes osteoclastogenesis and enhances osteogenesis of Ti implants, and can serve as a potent molecule for the development of good implantation. - Highlights: • Chitosan gold nanoparticles were conjugated with IGFBP-3 and coated onto surface of the titanium implants for gene delivery to bone. • Implants were inserted in rat mandible for 4 weeks. • Parameters studied: histopathology and radiology. �

Insulin-like growth factor binding protein-3 affects osteogenic efficacy on dental implants in rat mandible

International Nuclear Information System (INIS)

Bhattarai, Govinda; Lee, Young-Hee; Lee, Min-Ho; Park, Il-Song; Yi, Ho-Keun

2015-01-01

Insulin like growth factor binding protein-3 (IGFBP-3) in bone cells and its utilization in dental implants have not been well studied. The aim of this study was to determine the osteogenic efficacy of chitosan gold nanoparticles (Ch-GNPs) conjugated with IGFBP-3 coated titanium (Ti) implants. Ch-GNPs were conjugated with IGFBP-3 plasmid DNA through a coacervation process. Conjugation was cast over Ti surfaces, and cells were seeded on coated surfaces. For in vitro analysis the expression of different proteins was analyzed by immunoblotting. For in vivo analysis, Ch-GNP/IGFBP-3 coated implants were installed in rat mandibles. Four weeks post-implantation, mandibles were examined by microcomputed tomography (μCT), immunohistochemistry, hematoxylin & eosin and tartrate resistance acid phosphatase staining. In vitro overexpressed Ch-GNP/IGFBP-3 coated Ti surfaces was associated with activation of extracellular signal related kinase (ERK), inhibition of the stress activated protein c-Jun N-terminal kinase (JNK) and enhanced bone morphogenetic protein (BMP)-2 and 7 compared to control. Further, in vivo, Ch-GNP/IGFBP-3 coated implants were associated with inhibition of implant induced osteoclastogenesis molecules, receptor activator of nuclear factor kappa-B ligand (RANKL) and enhanced expression of osteogenic molecules including BMP2/7 and osteopontin (OPN). The μCT analysis demonstrated that IGFBP-3 increased the volume of newly formed bone surrounding the implants compared to control (n = 5; p < 0.05). These results support the view that IGFBP-3 overexpression diminishes osteoclastogenesis and enhances osteogenesis of Ti implants, and can serve as a potent molecule for the development of good implantation. - Highlights: • Chitosan gold nanoparticles were conjugated with IGFBP-3 and coated onto surface of the titanium implants for gene delivery to bone. • Implants were inserted in rat mandible for 4 weeks. • Parameters studied: histopathology and radiology. �

Robust embryo identification using first polar body single nucleotide polymorphism microarray-based DNA fingerprinting.

Science.gov (United States)

Treff, Nathan R; Su, Jing; Kasabwala, Natasha; Tao, Xin; Miller, Kathleen A; Scott, Richard T

2010-05-01

This study sought to validate a novel, minimally invasive system for embryo tracking by single nucleotide polymorphism microarray-based DNA fingerprinting of the first polar body. First polar body-based assignments of which embryos implanted and were delivered after multiple ET were 100% consistent with previously validated embryo DNA fingerprinting-based assignments. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Comparison of metal concentrations in rat tibia tissues with various metallic implants.

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Okazaki, Yoshimitsu; Gotoh, Emiko; Manabe, Takeshi; Kobayashi, Kihei

2004-12-01

To compare metal concentrations in tibia tissues with various metallic implants, SUS316L stainless steel, Co-Cr-Mo casting alloy, and Ti-6Al-4V and V-free Ti-15Zr-4Nb-4Ta alloys were implanted into the rat tibia for up to 48 weeks. After the implant was removed from the tibia by decalcification, the tibia tissues near the implant were lyophilized. Then the concentrations of metals in the tibia tissues by microwave acid digestion were determined by inductively coupled plasma-mass spectrometry. Fe concentrations were determined by graphite-furnace atomic absorption spectrometry. The Fe concentration in the tibia tissues with the SUS316L implant was relatively high, and it rapidly increased up to 12 weeks and then decreased thereafter. On the other hand, the Co concentration in the tibia tissues with the Co-Cr-Mo implant was lower, and it increased up to 24 weeks and slightly decreased at 48 weeks. The Ni concentration in the tibia tissues with the SUS316L implant increased up to 6 weeks and then gradually decreased thereafter. The Cr concentration tended to be higher than the Co concentration. This Cr concentration linearly increased up to 12 weeks and then decreased toward 48 weeks in the tibia tissues with the SUS316L or Co-Cr-Mo implant. Minute quantities of Ti, Al and V in the tibia tissues with the Ti-6Al-4V implant were found. The Ti concentration in the tibia tissues with the Ti-15Zr-4Nb-4Ta implant was lower than that in the tibia tissues with the Ti-6Al-4V implant. The Zr, Nb and Ta concentrations were also very low. The Ti-15Zr-4Nb-4Ta alloy with its low metal release in vivo is considered advantageous for long-term implants.

Titanium diffusion in shinbone of rats with osseointegrated implants.

Science.gov (United States)

Grenón, Miriam S; Robledo, José; Ibáñez, Juan Carlos; Sánchez, Héctor J

2016-11-01

Dental implants are composed of commercially pure Ti (which is actually an alloy of titanium, and minor or trace components such as aluminium and vanadium). When the implant is inserted, its surface undergoes a number of chemical and mechanical processes, releasing particles of titanium to the medium. The metabolism of free ions of titanium is uncertain; the uptaking processes in the body are not well known, nor their toxic dose. In addition, physical properties of newly formed bone, such as diffusivity and activation energy, are scarce and rarely studied. In this study, we analysed the diffusion of titanium in the titanium-implanted shinbones of six adult male Wistar rats by spatially resolved micro x-ray fluorescence. The measurements were carried out at the microfluorescence station of the x-ray fluorescence (XRF) beamline of the Brazilian synchrotron facility LNLS (from Portuguese 'Laboratorio Nacional de Luz Sincrotron'). For each sample, XRF spectra were taken by linear scanning in area near the new bone formed around the Ti implant. The scanning line shows a clear effect of titanium diffusion whereas calcium intensity presents a different behaviour. Moreover, a clear correlation among the different structures of bones is observed in the Ti and Ca intensities. The results obtained in these measurements may allow determining quantitatively the parameters of diffusion rates and other physical properties of new bone like diffusion coefficients. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

Change in ATP-ase activity and transport of rna of liver cell nuclei of pregnant rats and embryos following irradiation

International Nuclear Information System (INIS)

Shamsutdinova, G.T.; Mirkhamidova, P.; Ibragimkhodzhaeva, M.P.; Mirakhmedov, A.K.

1990-01-01

The effect of radiation on ATP-ase activity in rat liver nuclei and RNA transport of isolated liver nuclei in vitro is studied. It is shown that irradiation changes RNA transport from isolated liver cell nuclei of maternal organism and embryos. Irradiation during prefetus and fetus periods changes ATP-ase activity of embryon and maternal organism nuclei

Influence of irradiation (Co{sub 6}0) in pre-implant rabbits embryos: effect on blastocyst diameters and embryos smaller than 2 mm; Influencia da radiacao ionizante (bomba de cobalto) em embrioes de coelha na fase de pre-implantacao: efeito no diametro das blastulas e embrioes com menos de 2mm

Energy Technology Data Exchange (ETDEWEB)

Approbato, Mario S.; Oliveira Moura, Katia K.V. de; Souza Florencio, Rodopiano de; Cunha Junior, Carlos; Garcia, Ricardo; Faria, Renato S.; Benedetti, Leonardo N.; Goulart, Flamarion B.

1995-06-01

We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), in three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses, 5 c Gy and 10 c Gy. Six rabbits (36 blastocysts) were used as controls. The matching instant was the zero hour. Exactly six days after ({+-} 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. The embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryos parameters were studied: diameter growth; percentage of embryos smaller than 2 mm. We observed that only the irradiation time influenced the blastocysts diameter (no irradiation dose). There was no relation between percentage of embryos smaller than 2 mm and the irradiation. (author). 11 refs., 2 figs.

Single embryo transfer and IVF/ICSI outcome: a balanced appraisal.

Science.gov (United States)

Gerris, Jan M R

2005-01-01

This review considers the value of single embryo transfer (SET) to prevent multiple pregnancies (MP) after IVF/ICSI. The incidence of MP (twins and higher order pregnancies) after IVF/ICSI is much higher (approximately 30%) than after natural conception (approximately 1%). Approximately half of all the neonates are multiples. The obstetric, neonatal and long-term consequences for the health of these children are enormous and costs incurred extremely high. Judicious SET is the only method to decrease this epidemic of iatrogenic multiple gestations. Clinical trials have shown that programmes with >50% of SET maintain high overall ongoing pregnancy rates ( approximately 30% per started cycle) while reducing the MP rate to select patients suitable for SET and embryos with a high putative implantation potential. The typical patient suitable for SET is young (aged Embryo selection is performed using one or a combination of embryo characteristics. Available evidence suggests that, for the overall population, day 3 and day 5 selection yield similar results but better than zygote selection results. Prospective studies correlating embryo characteristics with documented implantation potential, utilizing databases of individual embryos, are needed. The application of SET should be supported by other measures: reimbursement of IVF/ICSI (earned back by reducing costs), optimized cryopreservation to augment cumulative pregnancy rates per oocyte harvest and a standardized format for reporting results. To make SET the standard of care in the appropriate target group, there is a need for more clinical studies, for intensive counselling of patients, and for an increased sense of responsibility in patients, health care providers and health insurers.

Folic acid and safflower oil supplementation interacts and protects embryos from maternal diabetes-induced damage.

Science.gov (United States)

Higa, R; Kurtz, M; Mazzucco, M B; Musikant, D; White, V; Jawerbaum, A

2012-05-01

Maternal diabetes increases the risk of embryo malformations. Folic acid and safflower oil supplementations have been shown to reduce embryo malformations in experimental models of diabetes. In this study we here tested whether folic acid and safflower oil supplementations interact to prevent embryo malformations in diabetic rats, and analyzed whether they act through the regulation of matrix metalloproteinases (MMPs), their endogenous inhibitors (TIMPs), and nitric oxide (NO) and reactive oxygen species production. Diabetes was induced by streptozotocin administration prior to mating. From Day 0.5 of pregnancy, rats did or did not receive folic acid (15 mg/kg) and/or a 6% safflower oil-supplemented diet. Embryos and decidua were explanted on Day 10.5 of gestation for further analysis of embryo resorptions and malformations, MMP-2 and MMP-9 activities, TIMP-1 and TIMP-2 levels, NO production and lipid peroxidation. Maternal diabetes induced resorptions and malformations that were prevented by folic acid and safflower oil supplementation. MMP-2 and MMP-9 activities were increased in embryos and decidua from diabetic rats and decreased with safflower oil and folic acid supplementations. In diabetic animals, the embryonic and decidual TIMPs were increased mainly with safflower oil supplementation in decidua and with folic acid in embryos. NO overproduction was decreased in decidua from diabetic rats treated with folic acid alone and in combination with safflower oil. These treatments also prevented increases in embryonic and decidual lipid peroxidation. In conclusion, folic acid and safflower oil supplementations interact and protect the embryos from diabetes-induced damage through several pathways related to a decrease in pro-inflammatory mediators.

Plasma concentrations and placental immunostaining of interleukin-10 and tumornecrosis factor-α as predictors of alterations in the embryo-fetal organism and the placental development of diabetic rats

Directory of Open Access Journals (Sweden)

Y.K. Sinzato

2011-03-01

Full Text Available Interleukin-10 (IL-10 appears to be the key cytokine for the maintenance of pregnancy and inhibits the secretion of inflammatory cytokines such as tumor necrosis factor-α (TNF-α. However, there are no studies evaluating the profile of these cytokines in diabetic rat models. Thus, our aim was to analyze IL-10 and TNF-α immunostaining in placental tissue and their respective concentrations in maternal plasma during pregnancy in diabetic rats in order to determine whether these cytokines can be used as predictors of alterations in the embryo-fetal organism and in placental development. These parameters were evaluated in non-diabetic (control; N = 15 and Wistar rats with streptozotocin (STZ-induced diabetes (N = 15. At term, the dams (100 days of life were killed under anesthesia and plasma and placental samples were collected for IL-10 and TNF-α determinations by ELISA and immunohistochemistry, respectively. The reproductive performance was analyzed. Plasma IL-10 concentrations were reduced in STZ rats compared to controls (7.6 ± 4.5 vs 20.9 ± 8.1 pg/mL. The placental scores of immunostaining intensity did not differ between groups (P > 0.05. Prevalence analysis showed that the IL-10 expression followed TNF-α expression, showing a balance between them. STZ rats also presented impaired reproductive performance and reduced plasma IL-10 levels related to damage during early embryonic development. However, the increased placental IL-10 as a compensatory mechanism for the deficit of maternal regulation permitted embryo development. Therefore, the data suggest that IL-10 can be used as a predictor of changes in the embryo-fetal organism and in placental development in pregnant diabetic rats.

A randomized clinical trial to evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium for in vitro fertilization

DEFF Research Database (Denmark)

Ziebe, Søren; Loft, Anne; Povlsen, Betina B

2013-01-01

To evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium on ongoing implantation rate (OIR).......To evaluate the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) in embryo culture medium on ongoing implantation rate (OIR)....

Genotoxicity determinations of coriander drop and extract of Coriander Sativum cultured fibroblast of rat embryo by comet assay

International Nuclear Information System (INIS)

Heibatullah, K.; Marzieh, P.; Arefeh, I.; Ebrahim, M.

2008-01-01

The single cell gel electrophoresis (SCGE) or comet assay is a quick, simple and sensitive technique for measuring DNA damage in cell nucleus. It is well known that medicinal herbs play an important role in the life of human beings, thus it is essential to determine their safety as public health is concerned. In this study the genotoxicity of Coriander drop, herbal pharmaceutical product, and the extract of Coriander sativum were examined in cultured fibroblast of rat embryo using comet assay. The thirteen to fifteen days old rat embryos were lysed with tripsin and after certain steps it was centrifuged and then cultured. After three to five passages, different concentrations of each product were applied to the fibroblasts. Lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed with fluorescence microscope. In the test groups the results indicated that coriander drop at different doses showed some fragmentation of DNA but this damage as a result was deemed to be not significant. However, in the case of Coriander sativum extract the results showed no mutagenic effects in comparison with the positive control group (p<0.05). In conclusion, these herbal products did not show any magnetic effect according to our test, but further genotoxicity assays are recommended. (author)

Effect of Bacterial Endotoxins on Superovulated Mouse Embryos In Vivo: Is CSF-1 Involved in Endotoxin-Induced Pregnancy Loss?

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Yogesh Kumar Jaiswal

2006-01-01

Full Text Available Mammalian embryonic development is regulated by several cytokines and growth factors from embryonic or maternal origins. Since CSF-1 plays important role in embryonic development and implantation, we investigated its role in gram-negative bacterial LPS-induced implantation failure. The effect of LPS on normal (nonsuperovulated and superovulated in vivo-produced embryos was assessed by signs of morphological degeneration. A significantly similar number of morphologically degenerated embryos recovered from both nonsuperovulated and superovulated LPS treated animals on day 2.5 of pregnancy onwards were morphologically and developmentally abnormal as compared to their respective controls (P < .001. Normal CSF-1 expression level and pattern were also altered through the preimplantation period in the mouse embryos and uterine horns after LPS treatment. This deviation from the normal pattern and level of CSF-1 expression in the preimplantation embryos and uterine tissues suggest a role for CSF-1 in LPS-induced implantation failure.

Embryogenesis-promoting factors in rat serum.

Science.gov (United States)

Katoh, M; Kimura, R; Shoji, R

1998-06-15

Regarding whole rat embryo cultures in vitro, rat serum as a culture medium is known to support the normal growth of rat embryos in the organogenesis phase. The purpose of the present study was to isolate the embryogenesis-promoting factors from rat serum as a first step in the development of a defined serum-free medium for a whole embryo culture system. Pooled rat serum after heat inactivation was fractionated into three major peaks (frA, containing a region of void volume, frB, and frC) by gel filtration. The 9.5-day rat embryos that were cultivated for 48 hr in essential salt medium containing frB (with a molecular size range of 100-500 kDa) revealed normal growth. Three proteins (27 kDa, 76 kDa, and 190 kDa) that had the embryogenesis-promoting effects were isolated from 3-hr delayed centrifuged rat serum by the ion exchange chromatography. The 76-kDa protein was found to be rat transferrin by immunoblotting. The 27-kDa protein was identified as apo-AI (the major apoprotein of high-density lipoprotein) by immunoblotting. High-density lipoprotein obtained from pooled rat serum by a NaBr density gradient ultracentrifugation was found to have a positive effect on embryogenesis. The 10-kDa protein was also identified as alpha 1-inhibitor 3 by immunoblotting. In addition, the embryogenesis-promoting effect of the fraction containing 27-kDa and 190-kDa proteins declined within a short period of storage at -20 degrees C. This decrease was countered by supplementing its fraction (D-2) with albumin isolated from rat serum. These results in the present study suggest that transferrin, high-density lipoprotein, and alpha 1-inhibitor 3 in rat serum may be embryogenesis-promoting factors, and that albumin appeared to play a role in the embryogenesis of rat embryos in whole embryo cultures.

Utilizing a Rat Delayed Implantation Model to Teach Integrative Endocrinology and Reproductive Biology

Science.gov (United States)

Geisert, Rodney D.; Smith, Michael F.; Schmelzle, Amanda L.; Green, Jonathan A.

2018-01-01

In this teaching laboratory, the students are directed in an exercise that involves designing and performing an experiment to determine estrogen's role in regulating delayed implantation (diapause) in female rats. To encourage active participation by the students, a discussion question is provided before the laboratory exercise in which each…

Is preimplantation genetic diagnosis the ideal embryo selection method in aneuploidy screening?

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Levent Sahin

2014-10-01

Full Text Available To select cytogenetically normal embryos, preimplantation genetic diagnosis (PGD aneuploidy screening (AS is used in numerous centers around the world. Chromosomal abnormalities lead to developmental problems, implantation failure, and early abortion of embryos. The usefulness of PGD in identifying single-gene diseases, human leukocyte antigen typing, X-linked diseases, and specific genetic diseases is well-known. In this review, preimplantation embryo genetics, PGD research studies, and the European Society of Human Reproduction and Embryology PGD Consortium studies and reports are examined. In addition, criteria for embryo selection, technical aspects of PGD-AS, and potential noninvasive embryo selection methods are described. Indications for PGD and possible causes of discordant PGD results between the centers are discussed. The limitations of fluorescence in situ hybridization, and the advantages of the array comparative genomic hybridization are included in this review. Although PGD-AS for patients of advanced maternal age has been shown to improve in vitro fertilization outcomes in some studies, to our knowledge, there is not sufficient evidence to use advanced maternal age as the sole indication for PGD-AS. PGD-AS might be harmful and may not increase the success rates of in vitro fertilization. At the same time PGD, is not recommended for recurrent implantation failure and unexplained recurrent pregnancy loss.

Who abandons embryos after IVF?

LENUS (Irish Health Repository)

Walsh, A P H

2010-04-01

This investigation describes features of in vitro fertilisation (IVF) patients who never returned to claim their embryos following cryopreservation. Frozen embryo data were reviewed to establish communication patterns between patient and clinic; embryos were considered abandoned when 1) an IVF patient with frozen embryo\\/s stored at our facility failed to make contact with our clinic for > 2 yrs and 2) the patient could not be located after a multi-modal outreach effort was undertaken. For these patients, telephone numbers had been disconnected and no forwarding address was available. Patient, spouse and emergency family contact\\/s all escaped detection efforts despite an exhaustive public database search including death records and Internet directory portals. From 3244 IVF cycles completed from 2000 to 2008, > or = 1 embryo was frozen in 1159 cases (35.7%). Those without correspondence for > 2 yrs accounted for 292 (25.2%) patients with frozen embryos; 281 were contacted by methods including registered (signature involving abandoned embryos did not differ substantially from other patients. The goal of having a baby was achieved by 10\\/11 patients either by spontaneous conception, adoption or IVF. One patient moved away with conception status unconfirmed. The overall rate of embryo abandonment was 11\\/1159 (< 1%) in this IVF population. Pre-IVF counselling minimises, but does not totally eliminate, the problem of abandoned embryos. As the number of abandoned embryos from IVF accumulates, their fate urgently requires clarification. We propose that clinicians develop a policy consistent with relevant Irish Constitutional provisions to address this medical dilemma.

Experiment K-313: Rat and quail ontogenesis

Science.gov (United States)

Keefe, J. R.

1981-01-01

The potential effects of spaceflight on the processes of mammalian fertilizaton, implantation and embryonic development are investigated. Five female and two male rats were placed together on Day 2 of the flight. By R+17, it was determined that both flight and synchronous females were not carrying normal pregnancies and three of the flight animals were laparotomized. The uterus and ovaries were processed for microscopic analyses. The two remaining flight females were allowed to recover from the exploratory operation, rebred with flight males and delivered normal litters. As a control for potential transplacental effects that might be interpreted as direct spaceflight effects, a series of fertilized Japanese quail (Coturnix japonica) eggs was flown on Cosmos 1129. Although all of the eggs were adversely impacted by an inflight failure of the incubator humidifier on flight Day 13, several embryos were able to progress to a developmental stage equivalent to that of a control 10-12 Day embryo.

Simple Genome Editing of Rodent Intact Embryos by Electroporation.

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Takehito Kaneko

Full Text Available The clustered regularly interspaced short palindromic repeat (CRISPR/CRISPR-associated (Cas system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique for animal knockout system by electroporation (TAKE method is a simple and effective technology that produces knockout rats by introducing endonuclease mRNAs into intact embryos using electroporation. Using TAKE method and CRISPR/Cas system, the present study successfully produced knockout and knock-in mice and rats. The mice and rats derived from embryos electroporated with Cas9 mRNA, gRNA and single-stranded oligodeoxynucleotide (ssODN comprised the edited targeted gene as a knockout (67% of mice and 88% of rats or knock-in (both 33%. The TAKE method could be widely used as a powerful tool to produce genetically modified animals by genome editing.

The effect of endometrial scratch on natural-cycle cryopreserved embryo transfer outcomes: a randomized controlled study.

Science.gov (United States)

Mak, Jennifer Sze Man; Chung, Cathy Hoi Sze; Chung, Jacqueline Pui Wah; Kong, Grace Wing Shan; Saravelos, Sotirios H; Cheung, Lai Ping; Li, Tin-Chiu

2017-07-01

The benefit of endometrial scratch (ES) prior to embryo transfer is controversial. Systemic analysis has confirmed its potential benefit, especially in women with repeated IVF failures, yet most studies have focused on fresh embryo transfer, and its effect on vitrified-warmed embryo transfer (FET) cycles is yet to be explored. We hereby present our prospective, double-blind, randomized controlled study on the evaluation of the implantation and pregnancy rate after ES prior to natural-cycle FET. A total of 299 patients underwent natural-cycle FET and were randomized to receive ES (n = 115) or endocervical manipulation as control (n = 114) prior to FET cycle, and a total of 196 patients had embryo transfer (93 patients in each group). Our study showed no significant difference in the implantation and pregnancy rate, as well as the clinical and ongoing pregnancy or live birth rates between the two groups. It appears that ES does not have any beneficial effect on an unselected group of women undergoing FET in natural cycles. Further studies on its effect in women with recurrent implantation failure after IVF are warranted. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

IVF/ICSI outcomes after culture of human embryos at low oxygen tension: a meta-analysis

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Gomes Sobrinho David B

2011-11-01

Full Text Available Abstract Background Improved pregnancy, implantation, and birth rates have been reported after the use of reduced O2 concentration during embryo culture, mainly due to a reduction of the cumulative detrimental effects of reactive oxygen species. However, some studies have failed to report any positive effects. The objective of this meta-analysis was to evaluate the effect of a low-O2 environment on IVF/intracytoplasmic sperm injection (ICSI outcomes. Methods All available published and ongoing randomised trials that compared the effects of low (~5%; OC~5 and atmospheric (~20%; OC~20 oxygen concentrations on IVF/ICSI outcomes were included. Search strategies included online surveys of databases from 1980 to 2011. The outcomes measured were fertilisation rate, implantation rate and ongoing pregnancy rates. The fixed effects model was used to calculate the odds ratio. Results Seven studies were included in this analysis. The pooled fertilisation rate did not differ significantly (P = 0.54 between the group of oocytes cultured at low O2 tension and the group at atmospheric O2 tension. Concerning all cycles, the implantation (P = 0.06 and ongoing pregnancy (P = 0.051 rates were not significantly different between the group receiving transferred sets containing only OC~5 embryos and the group receiving transferred sets with only OC~20 embryos. In a meta-analysis performed for only those trials in which embryos were transferred on day 2/3, implantation (P = 0.63 and ongoing pregnancy (P = 0.19 rates were not significantly different between the groups. In contrast, when a meta-analysis was performed using only trials in which embryos were transferred on days 5 and 6 (at the blastocyst stage, the group with transferred sets of only OC~5 embryos showed a statistically significantly higher implantation rate (P = 0.006 than the group receiving transferred sets with only OC~20 embryos, although the ongoing pregnancy (P = 0.19 rates were not significantly

Systemic Assessment of Calcium and Phosphorus Level after Implantation of Porous Iron in Rats

Science.gov (United States)

Siallagan, S. F.; Amelia, F.; Utami, N. D.; Ulum, M. F.; Boediono, A.; Estuningsih, S.; Hermawan, H.; Noviana, D.

2017-07-01

One of important aspects in bone healing process is physiological level of calcium (Ca), and phosphorus (P) that can be altered by implantation of biodegradable porous iron. Therefore, this study aims to investigate the concentration of Ca, P and Ca/P ratio in the peripheral blood during the implantation period up to 4 months. Forty adult male Sprague Dawley rats were used and divided into 3 groups receiving different pore size of iron implants (pore size 450, 580, 800μm) and one group of sham. The implants (5x2x0.5mm) were inserted into flat bone defects at latero-medial of femoral bone. Blood sample was taken from ventral tail artery before and after 4 month of implantation. Calcium and P concentrations in the blood were determined by BA-88A Semi-Auto Chemistry Analyzer. Results showed that concentration of Ca and P are slightly higher after implantation than before implantation, except for the 450μm group. The Ca/P ratio before and after implantation was increased in the sham group, and decreased in the 450 and 800μm groups. Concentration of Ca, P and Ca/P ratio insignificantly change between before and 4 months after surgery in some groups.

Intramyocardial implantation of differentiated rat bone marrow mesenchymal stem cells enhanced by TGF-β1 improves cardiac function in heart failure rats

Energy Technology Data Exchange (ETDEWEB)

Lv, Y. [Department of Histology and Embryology, Hebei Medical University, Shijiazhuang, Hebei (China); Liu, B. [Department of Pathology, the First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei (China); Wang, H.P. [Department of Histology and Embryology, Hebei North University, Zhangjiakou, Hebei (China); Zhang, L. [Department of Histology and Embryology, Hebei Medical University, Shijiazhuang, Hebei (China)

2016-05-31

The present study tested the hypotheses that i) transforming growth factor beta 1 (TGF-β1) enhances differentiation of rat bone marrow mesenchymal stem cells (MSCs) towards the cardiomyogenic phenotype and ii) intramyocardial implantation of the TGF-β1-treated MSCs improves cardiac function in heart failure rats. MSCs were treated with different concentrations of TGF-β1 for 72 h, and then morphological characteristics, surface antigens and mRNA expression of several transcription factors were assessed. Intramyocardial implantation of these TGF-β1-treated MSCs to infarcted heart was also investigated. MSCs were initially spindle-shaped with irregular processes. On day 28 after TGF-β1 treatment, MSCs showed fusiform shape, orientating parallel with one another, and were connected with adjoining cells forming myotube-like structures. Immunofluorescence revealed the expression of cardiomyocyte-specific proteins, α-sarcomeric actin and troponin T, in these cells. The mRNA expression of GATA4 and Nkx2.5 genes was slightly increased on day 7, enhanced on day 14 and decreased on day 28 while α-MHC gene was not expressed on day 7, but expressed slightly on day 14 and enhanced on day 28. Transmission electron microscopy showed that the induced cells had myofilaments, z line-like substances, desmosomes, and gap junctions, in contrast with control cells. Furthermore, intramyocardial implantation of TGF-β1-treated MSCs to infarcted heart reduced scar area and increased the number of muscle cells. This structure regeneration was concomitant with the improvement of cardiac function, evidenced by decreased left ventricular end-diastolic pressure, increased left ventricular systolic pressure and increased maximal positive pressure development rate. Taken together, these results indicate that intramyocardial implantation of differentiated MSCs enhanced by TGF-β1 improved cardiac function in heart failure rats.

Intramyocardial implantation of differentiated rat bone marrow mesenchymal stem cells enhanced by TGF-β1 improves cardiac function in heart failure rats

International Nuclear Information System (INIS)

Lv, Y.; Liu, B.; Wang, H.P.; Zhang, L.

2016-01-01

The present study tested the hypotheses that i) transforming growth factor beta 1 (TGF-β1) enhances differentiation of rat bone marrow mesenchymal stem cells (MSCs) towards the cardiomyogenic phenotype and ii) intramyocardial implantation of the TGF-β1-treated MSCs improves cardiac function in heart failure rats. MSCs were treated with different concentrations of TGF-β1 for 72 h, and then morphological characteristics, surface antigens and mRNA expression of several transcription factors were assessed. Intramyocardial implantation of these TGF-β1-treated MSCs to infarcted heart was also investigated. MSCs were initially spindle-shaped with irregular processes. On day 28 after TGF-β1 treatment, MSCs showed fusiform shape, orientating parallel with one another, and were connected with adjoining cells forming myotube-like structures. Immunofluorescence revealed the expression of cardiomyocyte-specific proteins, α-sarcomeric actin and troponin T, in these cells. The mRNA expression of GATA4 and Nkx2.5 genes was slightly increased on day 7, enhanced on day 14 and decreased on day 28 while α-MHC gene was not expressed on day 7, but expressed slightly on day 14 and enhanced on day 28. Transmission electron microscopy showed that the induced cells had myofilaments, z line-like substances, desmosomes, and gap junctions, in contrast with control cells. Furthermore, intramyocardial implantation of TGF-β1-treated MSCs to infarcted heart reduced scar area and increased the number of muscle cells. This structure regeneration was concomitant with the improvement of cardiac function, evidenced by decreased left ventricular end-diastolic pressure, increased left ventricular systolic pressure and increased maximal positive pressure development rate. Taken together, these results indicate that intramyocardial implantation of differentiated MSCs enhanced by TGF-β1 improved cardiac function in heart failure rats

Extensive review of fish embryo acute toxicities for the prediction of GHS acute systemic toxicity categories.

Science.gov (United States)

Scholz, Stefan; Ortmann, Julia; Klüver, Nils; Léonard, Marc

2014-08-01

Distribution and marketing of chemicals require appropriate labelling of health, physical and environmental hazards according to the United Nations global harmonisation system (GHS). Labelling for (human) acute toxicity categories is based on experimental findings usually obtained by oral, dermal or inhalative exposure of rodents. There is a strong societal demand for replacing animal experiments conducted for safety assessment of chemicals. Fish embryos are considered as alternative to animal testing and are proposed as predictive model both for environmental and human health effects. Therefore, we tested whether LC50s of the fish embryo acute toxicity test would allow effectively predicting of acute mammalian toxicity categories. A database of published fish embryo LC50 containing 641 compounds was established. For these compounds corresponding rat oral LD50 were identified resulting in 364 compounds for which both fish embryo LC50 and rat LD50 was available. Only a weak correlation of fish embryo LC50 and rat oral LD50 was obtained. Fish embryos were also not able to effectively predict GHS oral acute toxicity categories. We concluded that due to fundamental exposure protocol differences (single oral dose versus water-borne exposure) a reverse dosimetry approach is needed to explore the predictive capacity of fish embryos. Copyright © 2014 Elsevier Inc. All rights reserved.

Two-generation reproductive toxicity study of implanted depleted uranium (DU) in CD rats.

Science.gov (United States)

Arfsten, D P; Still, K R; Wilfong, E R; Johnson, E W; McInturf, S M; Eggers, J S; Schaeffer, D J; Bekkedal, M Y-V

2009-01-01

Depleted uranium (DU) munitions and armor plating have been used in several conflicts over the last 17 yr, including the Persian Gulf War and the Iraq War. Because of its effectiveness and availability, DU will continue to be used in military applications into the foreseeable future. There is much controversy over the use of DU in weapons and equipment because of its potential radiological and toxic hazards, and there is concern over the chronic adverse health effects of embedded DU shrapnel in war veterans and bystanders. This study evaluated the effects of long-term implantation of DU on the reproductive success of F0 generation adults and development and survival of subsequent F1 and F2 generations in a two-generation reproductive toxicity study. F0 generation Sprague-Dawley rats, 8 wk of age, were surgically implanted with 0, 4, 8, 12, or 20 DU pellets (1 x 2 mm). Inert implant control animals were implanted with 12 or 20 tantallum (Ta) pellets. The F0 generation was then mated at 120 d post DU implantation. In the F0 generation, when measured on postimplantation d 27 and 117, uranium was present in the urine of DU-implanted animals in a dose-dependent manner. F0 reproductive success was similar across treatment groups and the maternal retrieval test revealed no changes in maternal behavior. DU implantation exerted no effect on the survival, health, or well-being of the F0 generation. Necropsy results of F0 animals were negative with the exception of a marked inflammatory response surrounding the implanted DU pellets. For the F1 generation, measures of F1 development through postnatal day (PND) 20 were unremarkable and no gross abnormalities were observed in F1 offspring. No uranium was detected in whole-body homogenates of PND 4 or PND 20 pups. Necropsy findings of F1 PND 20 pups were negative and no instances of ribcage malformation were observed in F1 PND 20 pups. Body weight and body weight gain of F1 rats through PND 120 were similar across treatment

Dose–response analysis of phthalate effects on gene expression in rat whole embryo culture

Energy Technology Data Exchange (ETDEWEB)

Robinson, Joshua F. [National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Department of Toxicogenomics, Maastricht University, Maastricht (Netherlands); Verhoef, Aart; Beelen, Vincent A. van; Pennings, Jeroen L.A. [National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Piersma, Aldert H., E-mail: aldert.piersma@rivm.nl [National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Institute for Risk Assessment Sciences (IRAS), Utrecht University, Utrecht (Netherlands)

2012-10-01

The rat postimplantation whole embryo culture (WEC) model serves as a potential screening tool for developmental toxicity. In this model, cultured rat embryos are exposed during early embryogenesis and evaluated for morphological effects. The integration of molecular-based markers may lead to improved objectivity, sensitivity and predictability of WEC in assessing developmental toxic properties of compounds. In this study, we investigated the concentration-dependent effects of two phthalates differing in potency, mono(2-ethylhexyl) phthalate (MEHP) and monomethyl phthalate (MMP, less toxic), on the transcriptome in WEC to examine gene expression in relation with dysmorphogenesis. MEHP was more potent than MMP in inducing gene expression changes as well as changes on morphology. MEHP induced significant enrichment of cholesterol/lipid/steroid (CLS) metabolism and apoptosis pathways which was associated with developmental toxicity. Regulation of genes within CLS metabolism pathways represented the most sensitive markers of MEHP exposure, more sensitive than classical morphological endpoints. As shown in direct comparisons with toxicogenomic in vivo studies, alterations in the regulation of CLS metabolism pathways has been previously identified to be associated with developmental toxicity due to phthalate exposure in utero. Our results support the application of WEC as a model to examine relative phthalate potency through gene expression and morphological responses. Additionally, our results further define the applicability domain of the WEC model for developmental toxicological investigations. -- Highlights: ► We examine the effect of two phthalates on gene expression and morphology in WEC. ► MEHP is more potent than MMP in inducing gene expression changes and dysmorphogenesis. ► MEHP significantly disrupts cholesterol metabolism pathways in a dose-dependent manner. ► Specific phthalate-related mechanisms in WEC are relevant to mechanisms in vivo.

Injection of embryo culture supernatant to the endometrial cavity does not affect outcomes in IVF/ICSI or oocyte donation cycles: a randomized clinical trial.

Science.gov (United States)

Prapas, Yannis; Petousis, Stamatios; Panagiotidis, Yannis; Gullo, Giuseppe; Kasapi, Lia; Papadeothodorou, Achilleas; Prapas, Nikos

2012-06-01

To evaluate whether intrauterine injection of embryo culture supernatant before embryo transfer has any impact on pregnancy and implantation rates. A total of 400 cycles, of which 200 IVF/ICSI and 200 oocyte donor (OD), were randomly assigned to have their uterine cavity injected (group I) or not (group II). Primary endpoints to be studied were pregnancy and implantation rates. Clinical pregnancy rate per transfer (47.87%, 90/188 versus 48.45%, 94/194) based on transvaginal scan findings at 7 weeks of gestation and implantation rate (25.6% versus 26.5%) were similar in the two groups. The day of embryo transfer, day 3 or day 5, did not affect the final outcome. Injection of embryo culture supernatant into the uterine cavity, 30 min before the embryo transfer on either day 3 or 5, neither improves nor adversely affects the pregnancy rate in IVF/ICSI or oocyte donation cycles. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

ART culture conditions change the probability of mouse embryo gestation through defined cellular and molecular responses

NARCIS (Netherlands)

Schwarzer, Caroline; Esteves, Telma Cristina; Arau´zo-Bravo, Marcos J.; le Gac, Severine; Nordhoff, Verena; Schlatt, Stefan; Boiani, Michele

2012-01-01

Do different human ART culture protocols prepare embryos differently for post-implantation development? ... Our data promote awareness that human ART culture media affect embryo development. Effects reported here in the mouse may apply also in human, because no ART medium presently available on the

[Traditional and modern approaches to culture of preimplantation mammalian embryos in vitro].

Science.gov (United States)

Brusentsev, E Iu; Igonina, T N; Amstislavskiĭ, S Ia

2014-01-01

This review covers the basic principles and methods of in vitro culture of preimplantation mammalian embryos. The features of in vitro development of embryos of various species of animals with allowance for the composition of nutrient media are described, with special attention paid to those species that have traditionally been consideredas laboratory (i.e., mice, rats, and hamsters). The effects of suboptimal culturing conditions of preimplantation embryos on the formation of the phenotype of individuals developed from these embryos are discussed. New approaches to optimize the conditions of the development of preimplantation mammalian embryos in vitro are analyzed.

Comparison of rat and rabbit embryo-fetal developmental toxicity data for 379 pharmaceuticals: on the nature and severity of developmental effects (Critical Reviews in Toxicology)

Science.gov (United States)

Regulatory non-clinical safety testing of human pharmaceutical compounds typically requires embryo fetal developmental toxicity (EFDT) testing in two species, (one rodent and one non-rodent, usually the rat and the rabbit). The question has been raised whether under some conditio...

A novel embryo culture media supplement that improves pregnancy rates in mice.

Science.gov (United States)

Highet, A R; Bianco-Miotto, T; Pringle, K G; Peura, A; Bent, S; Zhang, J; Nottle, M B; Thompson, J G; Roberts, C T

2017-03-01

The preimplantation embryo in vivo is exposed to numerous growth factors in the female reproductive tract, which are not recapitulated in embryo culture media in vitro The IGF2 and plasminogen activator systems facilitate blastocyst development. We hypothesized that the addition of IGF2 in combination with urokinase plasminogen activator (uPA) and plasminogen could improve rates of blastocyst hatching and implantation in mice. B6BcF1 and CBAB6F2 mouse embryos were divided into one of four supplemented culture media treatment groups: (1) control (media only); (2) 12.5 nM IGF2; (3) 10 µg/mL uPA and 5 µg/mL plasminogen; or (4) a combination of IGF2, uPA and plasminogen treatments. Embryo development to blastocyst stage and hatching were assessed before transfer to pseudopregnant recipient females and implantation, pregnancy rates and postnatal growth were assessed. After 90.5 h of culture, IGF2 + U + P treatment increased the percentage of B6BcF1 embryos that were hatching/hatched and percentage developing to blastocyst stage compared with controls (P culture, IGF2, uPA and plasminogen supplementation of culture media can improve pregnancy success, but the effect of treatment is dependent on the mouse strain. © 2017 Society for Reproduction and Fertility.

Implanted, inductively-coupled, radiofrequency coils fabricated on flexible polymeric material: Application to in vivo rat brain MRI at 7 T

International Nuclear Information System (INIS)

Ginefri, J.C.; Poirier-Quinot, M.; Darrasse, L.; Rubin, A.; Tatoulian, M.; Woytasik, M.; Boumezbeur, F.; Djemai, B.; Lethimonnier, F.

2012-01-01

Combined with high-field MRI scanners, small implanted coils allow for high resolution imaging with locally improved SNR, as compared to external coils. Small flexible implantable coils dedicated to in vivo MRI of the rat brain at 7 T were developed. Based on the Multi-turn Transmission Line Resonator design, they were fabricated with a Teflon substrate using copper micro-molding process and a specific metal-polymer adhesion treatment. The implanted coils were made biocompatible by Polydimethylsiloxane (PDMS) encapsulation. The use of low loss tangent material achieves low dielectric losses within the substrate and the use of the PDMS layer reduces the parasitic coupling with the surrounding media. An implanted coil was implemented in a 7 T MRI system using inductive coupling and a dedicated external pick-up coil for signal transmission. In vivo images of the rat brain acquired with in plane resolution of (150 μm) 2 thanks to the implanted coil revealed high SNR near the coil, allowing for the visualization of fine cerebral structures. (authors)

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

Science.gov (United States)

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

2015-03-01

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

Effect of MeV energy He and N pre-implantation on the formation of porous silicon

International Nuclear Information System (INIS)

Manuaba, A.; Paszti, F.; Ortega, C.; Grosman, A.; Horvath, Z.E.; Szilagyi, E.; Khanh, N.Q.; Vickridge, I.

2001-01-01

The effects of MeV energy He and N pre-implantation of Si substrate on the structure of porous silicon formed by anodic etching were studied by measuring the depth profiles of 15 N decorating the pores walls. Radiation damage was recovered by annealing after the implantation. It was found that the He implant accelerates the etching process, probably due to the bubbles or the remaining lattice damage. At a dose of 8x10 16 ions/cm 2 the He containing layer was formed with a significantly enhanced porosity due to the contribution of the large-sized bubbles. At the highest dose of 32.5x10 16 ions/cm 2 flaking took place during the anodic etching. In contrast to He, N stopped the anodic etching at a depth of critical N concentration of ∼0.9 at.%. For the lowest implantation dose, where the peak concentration was below this limit, the pores propagate through the implanted layer with an enhanced speed

Histological study on acute inflammatory reaction to polyurethane-coated silicone implants in rats Estudo histológico da reação inflamatória aguda ao implante de silicone revestido com poliuretano em ratos

OpenAIRE

Paulo Roberto da Silva Mendes; Jorge Bins-Ely; Eduardo Arnaut dos Santos Lima; Zulmar Antonio Accioli de Vasconcellos; Armando José d'Acampora; Rodrigo d' Eça Neves

2008-01-01

PURPOSE: Evaluating histologically the silicone peri-implant coated by polyurethane inflammation associated to the use of anti-microbial and bacterial contamination. METHODS: It was used 35 Wistar rats. The animals were divided in seven groups: I - Control; II - implant cavity contamination with10 bacteria/ml; III - implant cavity contamination with 10 bacteria/ml; IV - implant cavity contamination with 10 bacteria/ml; V - identical contamination to group II and implant immersions in anti-mic...

Embryo implantation: Shedding light on the roles of ovarian ...

African Journals Online (AJOL)

Huda

Implantation is a crucial step in mammalian reproduction, as it is a gateway to further embryonic development and successful pregnancy. Successful implantation requires coordinated interactions between .... Interestingly, LIF expression is suppressed in stressed mice. .... a role, under the influence of sex hormones, in the.

Interspecific in vitro assay for the chimera-forming ability of human pluripotent stem cells.

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Masaki, Hideki; Kato-Itoh, Megumi; Umino, Ayumi; Sato, Hideyuki; Hamanaka, Sanae; Kobayashi, Toshihiro; Yamaguchi, Tomoyuki; Nishimura, Ken; Ohtaka, Manami; Nakanishi, Mahito; Nakauchi, Hiromitsu

2015-09-15

Functional assay limitations are an emerging issue in characterizing human pluripotent stem cells (PSCs). With rodent PSCs, chimera formation using pre-implantation embryos is the gold-standard assay of pluripotency (competence of progeny to differentiate into all three germ layers). In human PSCs (hPSCs), however, this can only be monitored via teratoma formation or in vitro differentiation, as ethical concerns preclude generation of human-human or human-animal chimeras. To circumvent this issue, we developed a functional assay utilizing interspecific blastocyst injection and in vitro culture (interspecies in vitro chimera assay) that enables the development and observation of embryos up to headfold stage. The assay uses mouse pre-implantation embryos and rat, monkey and human PSCs to create interspecies chimeras cultured in vitro to the early egg-cylinder stage. Intra- and interspecific chimera assays with rodent PSC lines were performed to confirm the consistency of results in vitro and in vivo. The behavior of chimeras developed in vitro appeared to recapitulate that of chimeras developed in vivo; that is, PSC-derived cells survived and were integrated into the epiblast of egg-cylinder-stage embryos. This indicates that the interspecific in vitro chimera assay is useful in evaluating the chimera-forming ability of rodent PSCs. However, when human induced PSCs (both conventional and naïve-like types) were injected into mouse embryos and cultured, some human cells survived but were segregated; unlike epiblast-stage rodent PSCs, they never integrated into the epiblast of egg-cylinder-stage embryos. These data suggest that the mouse-human interspecies in vitro chimera assay does not accurately reflect the early developmental potential/process of hPSCs. The use of evolutionarily more closely related species as host embryos might be necessary to evaluate the developmental potency of hPSCs. © 2015. Published by The Company of Biologists Ltd.

Analysis of metallic traces from the biodegradation of endomedullary AZ31 alloy temporary implants in rat organs after long implantation times.

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Bodelón, O G; Iglesias, C; Garrido, J; Clemente, C; Garcia-Alonso, M C; Escudero, M L

2015-08-04

AZ31 alloy has been tested as a biodegradable material in the form of endomedullary implants in female Wistar rat femurs. In order to evaluate the accumulation of potentially toxic elements from the biodegradation of the implant, magnesium (Mg), aluminium (Al), zinc (Zn), manganese (Mn) and fluorine (F) levels have been measured in different organs such as kidneys, liver, lungs, spleen and brain. Several factors that may influence accumulation have been taken into account: how long the implant has been in place, whether or not the bone is fractured, and the presence of an MgF2 protective coating on the implant. The main conclusions and the clinical relevance of the study have been that AZ31 endomedullary implants have a degradation rate of about 60% after 13 months, which is fully compatible with fracture consolidation. Neither bone fracture nor an MgF2 coating seems to influence the accumulation of trace elements in the studied organs. Aluminium is the only alloying element in this study that requires special attention. The increase in Al recovered from the sampled organs represents 3.95% of the amount contained in the AZ31 implant. Al accumulates in a statistically significant way in all the organs except the brain. All of this suggests that in long-term tests AZ31 may be a suitable material for osteosynthesis.

Investigation of Implantable Multi-Channel Electrode Array in Rat Cerebral Cortex Used for Recording

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Taniguchi, Noriyuki; Fukayama, Osamu; Suzuki, Takafumi; Mabuchi, Kunihiko

There have recently been many studies concerning the control of robot movements using neural signals recorded from the brain (usually called the Brain-Machine interface (BMI)). We fabricated implantable multi-electrode arrays to obtain neural signals from the rat cerebral cortex. As any multi-electrode array should have electrode alignment that minimizes invasion, it is necessary to customize the recording site. We designed three types of 22-channel multi-electrode arrays, i.e., 1) wide, 2) three-layered, and 3) separate. The first extensively covers the cerebral cortex. The second has a length of 2 mm, which can cover the area of the primary motor cortex. The third array has a separate structure, which corresponds to the position of the forelimb and hindlimb areas of the primary motor cortex. These arrays were implanted into the cerebral cortex of a rat. We estimated the walking speed from neural signals using our fabricated three-layered array to investigate its feasibility for BMI research. The neural signal of the rat and its walking speed were simultaneously recorded. The results revealed that evaluation using either the anterior electrode group or posterior group provided accurate estimates. However, two electrode groups around the center yielded poor estimates although it was possible to record neural signals.

Importance of Maternal Diabetes on the Chronological Deregulation of the Intrauterine Development: An Experimental Study in Rat

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Marcela Salazar García

2015-01-01

Full Text Available We investigated whether maternal diabetes induced in rats using streptozotocin (STZ on Day 5 of pregnancy affects the intrauterine developmental timeline. A total of 30 pregnant Sprague-Dawley diabetic rats (DRs and 20 control rats (CRs were used to obtain 21-day fetuses (F21 and newborn (NB pups. Gestational age, weight, and body size were recorded as were the maxillofacial morphometry and morphohistological characteristics of the limbs. In DRs, pregnancy continued for ∼1.7 days, and delivery occurred 23 days postcoitus (DPC. In this group, the number of pups was lower, and 13% had maxillofacial defects. F21 in the DR group had lower weights and were smaller; moreover, the morphological characteristics of the maxillofacial structures, derived from the neural crest, were discordant with their chronological gestational age, resembling 18- to 19-day-old fetuses. These deficiencies were counterbalanced in NB pups. We conclude that hyperglycemia, which results from maternal diabetes and precedes embryo implantation, deregulates the intrauterine developmental timeline, restricts embryo-fetal growth, and primarily delays the remodeling and maturation of the structures derived from neural crest cells.

Evidence of increased endometrial vascular permeability at the time of implantation in the short-nosed fruit bat, Cyanopterus sphinx.

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Pakrasi, Pranab Lal; Tiwari, Anjana

2007-09-01

Early embryonic development and implantation were studied in tropical short-nosed fruit bat Cyanopterus sphinx. We report preimplantation development and embryo implantation. Different stages of cleavage were observed in embryo by direct microscopic examination of fresh embryos after retrieving them either from the oviduct or the uterus at different days, up to the day of implantation. Generally, the embryos enter the uterus at the 8-cell stage. Embryonic development continued without any delay and blastocyst were formed showing attachment to the uterine epithelium at the mesometrial side of the uterus. A distinct blue band was formed in the uterus. The site of blastocyst attachment was visualized as a blue band following intravenous injection of pontamine blue. Implantation occurred 9+/-0.7 days after mating. This study reports that bat embryonic development can be studied like other laboratory animals and that this bat shows blue dye reaction, indicating the site and exact time of implantation. This blue dye reaction can be used to accurately find post-implantational delay. We prove conclusively that this species of tropical bat does not have any type of embryonic diapause.

Melatonin Promotes the In Vitro Development of Microinjected Pronuclear Mouse Embryos via Its Anti-Oxidative and Anti-Apoptotic Effects.

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Tian, Xiuzhi; Wang, Feng; Zhang, Lu; Ji, Pengyun; Wang, Jing; Lv, Dongying; Li, Guangdong; Chai, Menglong; Lian, Zhengxing; Liu, Guoshi

2017-05-05

CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats) combined with pronuclear microinjection has become the most effective method for producing transgenic animals. However, the relatively low embryo developmental rate limits its application. In the current study, it was observed that 10 -7 M melatonin is considered an optimum concentration and significantly promoted the in vitro development of murine microinjected pronuclear embryos, as indicated by the increased blastocyst rate, hatching blastocyst rate and blastocyst cell number. When these blastocysts were implanted into recipient mice, the pregnancy rate and birth rate were significantly higher than those of the microinjected control, respectively. Mechanistic studies revealed that melatonin treatment reduced reactive oxygen species (ROS) production and cellular apoptosis during in vitro embryo development and improved the quality of the blastocysts. The implantation of quality-improved blastocysts led to elevated pregnancy and birth rates. In conclusion, the results revealed that the anti-oxidative and anti-apoptotic activities of melatonin improved the quality of microinjected pronuclear embryos and subsequently increased both the efficiency of embryo implantation and the birth rate of the pups. Therefore, the melatonin supplementation may provide a novel alternative method for generating large numbers of transgenic mice and this method can probably be used in human-assisted reproduction and genome editing.

Thermal desorption of deuterium from polycrystalline nickel pre-implanted with helium

International Nuclear Information System (INIS)

Shi, S.Q.; Abramov, E.; Thompson, D.A.

1990-01-01

The thermal desorption technique has been used to study the trapping of deuterium atoms in high-purity polycrystalline nickel pre-implanted with helium for 1 x 10 19 to 5 x 10 20 ions/m 2 . The effect of post-implantation annealing at 703 K and 923 K on the desorption behavior was investigated. Measured values of the total amount of detrapped deuterium (Q T ) and helium concentration were used in a computer simulation of the desorption curve. It was found that the simulation using one or two discrete trap energies resulted in an inadequate fit between the simulated and the measured data. Both experimental and simulation results are explained using a stress-field trapping model. The effective binding energy, E b eff , was estimated to be in the range of 0.4-0.6 eV. Deuterium charging was found to stimulate a release of helium at a relatively low temperature

Pre- and Postoperative Binaural Unmasking for Bimodal Cochlear Implant Listeners.

Science.gov (United States)

Sheffield, Benjamin M; Schuchman, Gerald; Bernstein, Joshua G W

Cochlear implants (CIs) are increasingly recommended to individuals with residual bilateral acoustic hearing. Although new hearing-preserving electrode designs and surgical approaches show great promise, CI recipients are still at risk to lose acoustic hearing in the implanted ear, which could prevent the ability to take advantage of binaural unmasking to aid speech recognition in noise. This study examined the tradeoff between the benefits of a CI for speech understanding in noise and the potential loss of binaural unmasking for CI recipients with some bilateral preoperative acoustic hearing. Binaural unmasking is difficult to evaluate in CI candidates because speech perception in noise is generally too poor to measure reliably in the range of signal to noise ratios (SNRs) where binaural intelligibility level differences (BILDs) are typically observed (binaural benefit, 9 out of 10 listeners tested postoperatively had performance equal to or better than their best pre-CI performance. The listener who retained functional acoustic hearing in the implanted ear also demonstrated a preserved acoustic BILD postoperatively. Approximately half of the CI candidates in this study demonstrated preoperative binaural hearing benefits for audiovisual speech perception in noise. Most of these listeners lost their acoustic hearing in the implanted ear after surgery (using nonhearing-preservation techniques), and therefore lost access to this binaural benefit. In all but one case, any loss of binaural benefit was compensated for or exceeded by an improvement in speech perception with the CI. Evidence of a preoperative BILD suggests that certain CI candidates might further benefit from hearing-preservation surgery to retain acoustic binaural unmasking, as demonstrated for the listener who underwent hearing-preservation surgery. This test of binaural audiovisual speech perception in noise could serve as a diagnostic tool to identify CI candidates who are most likely to receive

Detection of SEA-type α-thalassemia in embryo biopsies by digital PCR

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Ta-Hsien Lee

2017-08-01

Conclusion: The SEA-type deletion in cultured embryos can be sensitively diagnosed with the digital PCR procedure in clinics. The adoption of this robust PGD method could prevent the implantation of IVF embryos that are destined to develop Bart's hydrops fetalis in a timely manner. The results also help inform future development of a standard digital PCR procedure for cost-effective PGD of α-thalassemia in a standard IVF clinic.

Interspecies chimera between primate embryonic stem cells and mouse embryos: monkey ESCs engraft into mouse embryos, but not post-implantation fetuses.

Science.gov (United States)

Simerly, Calvin; McFarland, Dave; Castro, Carlos; Lin, Chih-Cheng; Redinger, Carrie; Jacoby, Ethan; Mich-Basso, Jocelyn; Orwig, Kyle; Mills, Parker; Ahrens, Eric; Navara, Chris; Schatten, Gerald

2011-07-01

Unequivocal evidence for pluripotency in which embryonic stem cells contribute to chimeric offspring has yet to be demonstrated in human or nonhuman primates (NHPs). Here, rhesus and baboons ESCs were investigated in interspecific mouse chimera generated by aggregation or blastocyst injection. Aggregation chimera produced mouse blastocysts with GFP-nhpESCs at the inner cell mass (ICM), and embryo transfers (ETs) generated dimly-fluorescencing abnormal fetuses. Direct injection of GFP-nhpESCs into blastocysts produced normal non-GFP-fluorescencing fetuses. Injected chimera showed >70% loss of GFP-nhpESCs after 21 h culture. Outgrowths of all chimeric blastocysts established distinct but separate mouse- and NHP-ESC colonies. Extensive endogenous autofluorescence compromised anti-GFP detection and PCR analysis did not detect nhpESCs in fetuses. NhpESCs localize to the ICM in chimera and generate pregnancies. Because primate ESCs do not engraft post-implantation, and also because endogenous autofluorescence results in misleading positive signals, interspecific chimera assays for pluripotency with primate stem cells is unreliable with the currently available ESCs. Testing primate ESCs reprogrammed into even more naïve states in these inter-specific chimera assays will be an important future endeavor. Copyright © 2011 Elsevier B.V. All rights reserved.

Factors affecting the outcome of "endometrial scratch" in women with recurrent implantation failure.

Science.gov (United States)

Coughlan, Carol; Yuan, Xi; Demirol, Aygul; Ledger, William; Li, Tin Chiu

2014-01-01

To examine factors affecting the outcome of the endometrial scratch in women with recurrent implantation failure. A total of 57 eligible patients with a history of recurrent implantation failure underwent an endometrial biopsy in the luteal phase of the menstrual cycle in the month immediately preceding the embryo transfer cycle. The comparative group consisted of a retrospective cohort of 66 women with recurrent implantation failure but without endometrial biopsy. There were no significant differences between the intervention and control groups in terms of age, follicle-stimulating hormone (FSH), free androgen index, anti-Müllerian hormone, body mass index, the number of embryos transferred, and the number of embryo transfer cycles. The clinical pregnancy rate in the intervention group (53%) was significantly (p 10 IU/L. Women with a normal FSH are more likely to derive benefit from endometrial scratch.

Day 4 good morula embryo transfer provided compatible live birth rate with day 5 blastocyst embryo in fresh IVF/ET cycles.

Science.gov (United States)

Li, Ryh-Sheng; Hwu, Yuh-Ming; Lee, Robert Kuo-Kuang; Li, Sheng-Hsiang; Lin, Ming-Huei

2018-02-01

Embryo transfers during cleavage stage (day 2 or day 3) and blastocyst stages (day 5 or day 6) are common in current daily practice in fresh IVF/ET cycles. Data regarding transferring day 4 embryos, morula/compact stage, is still restricted and the grading system is also inconsistent, as between IVF clinics. This study provided a new detailed classification system for morula/compact stage embryos and compared successes rates between day 4 and day 5 ET. This was a retrospective study. A review of medical records from January 1st, 2013, to December 31st 2015, performed for all conventional insemination and ICSI cycles with a GnRH-antagonist protocol at the Infertility Division of MacKay Memorial Hospital in Taipei City, Taiwan. There were 427 cycles included in our study, 107 in study group (day 4 MET) and 320 in control group (day 5 BET). Pregnancy rates and live birth rate were compatible, as between morula embryo transfer (MET) and blastocyst embryo transfer (BET). The implantation rate (36.3% vs. 39.6%, respectively, p = 0.500), clinical pregnancy rate (49.5% vs. 51.9%, respectively, p = 0.737), and live birth rate (42.1% vs. 45.6%, respectively, p = 0.574) were statistically insignificant between groups. The term birth rate was statistically higher in the MET group than in the BET group (95.7% vs. 79.5%, respectively, p = 0.006). When the clinical outcomes between day 4 good MET and day 5 good BET were compared, the results were compatible. The implantation rate (48.8% vs. 41.1%, respectively, p = 0.335), clinical pregnancy rate (55.0% vs. 53.2%, respectively, p = 0.867), and live birth rate (47.5% vs. 47.1%, respectively, p = 1.000) showed no significant difference. The term birth rate was also higher in day 4 good MET group than in day 5 good BET group (100% vs. 78.3%, respectively, p = 0.025). In this study, we performed day 4 MET avoid BET on Sunday. The grading system we provided was more detailed for embryo selection and it was easier to

SU-F-T-44: A Comparison of the Pre-Plan, Intra-Operative Plan, and Post-Implant Dosimetry for a Prostate Implant Case Using Prefabricated Linear Polymer-Encapsulated Pd-103

International Nuclear Information System (INIS)

Sheu, R; Powers, A; McGee, H; Stock, R; Lo, Y

2016-01-01

Purpose: To investigate the reproducibility and limitations of Pd-103 prostate brachytherapy using fixed length linear sources (CivaString). Methods: An LDR prostate brachytherapy case which was preplanned on MR images with prefabricated linear polymer-encapsulated Pd-103 sources (CivaString) was studied and compared with ultrasound based intra-operative planning and CT based post-implant dosimetry. We evaluated the following parameters among the three studies: prostate geometry (volume and cross sectional area), needle position and alignment deviations, and dosimetry parameters (D90). Results: The prostate volumes and axial cross sectional areas at center of prostate were measured as 41.8, 39.3 and 36.8 cc, and 14.9, 14.3, and 11.3 respectively on pre-plan MR, inter-op US, and post-implant CT studies. The deviation of prostate volumes and axial cross sectional areas measured on pre-planning MR and intra-operative US were within 5%. 17 out of 19 pre-planned needles were positioned within 5mm (the template grid size). One needle location was adjusted intra-operatively and another needle was removed due to proximity to urethra. The needle pathways were not always parallel to the trans-rectal probe due to the flexibility of CivaString. The angle of deviation was up to 10 degrees. Two pairs of needles were exchanged to better fit the length of prostate at the time of implant. This resulted in a prostate D90 of 153.8 Gy (124%) and 131.4 Gy (106.7%) for intra-op and PID respectively. Conclusion: Preplanning is a necessary part of implants performed with prefabricated linear polymer sources. However, as is often the case, there were real-time deviations from the pre-plan. Intra-op planning provides the ability conform to anatomy at the time of implant. Therefore, we propose to develop a systematic way to order extra strings of different length to provide the flexibility to perform intra-operative planning with fixed length strands.

SU-F-T-44: A Comparison of the Pre-Plan, Intra-Operative Plan, and Post-Implant Dosimetry for a Prostate Implant Case Using Prefabricated Linear Polymer-Encapsulated Pd-103

Energy Technology Data Exchange (ETDEWEB)

Sheu, R; Powers, A; McGee, H; Stock, R; Lo, Y [Mount Sinai Medical Center, New York, NY (United States)

2016-06-15

Purpose: To investigate the reproducibility and limitations of Pd-103 prostate brachytherapy using fixed length linear sources (CivaString). Methods: An LDR prostate brachytherapy case which was preplanned on MR images with prefabricated linear polymer-encapsulated Pd-103 sources (CivaString) was studied and compared with ultrasound based intra-operative planning and CT based post-implant dosimetry. We evaluated the following parameters among the three studies: prostate geometry (volume and cross sectional area), needle position and alignment deviations, and dosimetry parameters (D90). Results: The prostate volumes and axial cross sectional areas at center of prostate were measured as 41.8, 39.3 and 36.8 cc, and 14.9, 14.3, and 11.3 respectively on pre-plan MR, inter-op US, and post-implant CT studies. The deviation of prostate volumes and axial cross sectional areas measured on pre-planning MR and intra-operative US were within 5%. 17 out of 19 pre-planned needles were positioned within 5mm (the template grid size). One needle location was adjusted intra-operatively and another needle was removed due to proximity to urethra. The needle pathways were not always parallel to the trans-rectal probe due to the flexibility of CivaString. The angle of deviation was up to 10 degrees. Two pairs of needles were exchanged to better fit the length of prostate at the time of implant. This resulted in a prostate D90 of 153.8 Gy (124%) and 131.4 Gy (106.7%) for intra-op and PID respectively. Conclusion: Preplanning is a necessary part of implants performed with prefabricated linear polymer sources. However, as is often the case, there were real-time deviations from the pre-plan. Intra-op planning provides the ability conform to anatomy at the time of implant. Therefore, we propose to develop a systematic way to order extra strings of different length to provide the flexibility to perform intra-operative planning with fixed length strands.

Effect of Pre-treatment with Moringa oleifera (Drumstick Leaves on Diabetogenesis Produced by Alloxan in Rats

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Shamsun Nahar

2015-07-01

Full Text Available Background: Medicinal plants constitute an important source of potential therapeutic agents for diabetes. Objective: In the study, we aimed to investigate the pre-treatment effect or preventive effects of Moringa oleifera (MO leaves on blood sugar of rats. Materials and method: This experimental study was carried out in the department of Pharmacology and Therapeutics of Sir Salimullah Medical College in collaboration with Bangladesh Council of Scientific and Industrial Research (BCSIR, Dhaka. A total 24 long Evans rats were included in this study and divided in to four groups. Hyperglycemia was induced on rats using alloxan (100 mg/kg body weight, intraperitioneally. Blood sample was collected from tail vein by tail tipping method. Pre-treatment effect or preventive role of Moringa oleifera (drumstick leaf powder on diabetogenesis produced by Alloxan in rats was tested by giving 50 mg/rat/day Moringa oleifera leaf powder for 14 days orally as pre-treatment along with standard rat feed. Then alloxan was administered intraperitoneally on 15th day of the experiment and 50mg/rat/day Moringa oleifera leaf powder was given for 7 days as post-treatment. Results: No significant effect of MO on blood glucose level was observed on normal rats and non significant hypoglycaemic effect was found in rats that were pretreated with MO. Conclusion: The present study suggests that Moringa oleifera leaf powder did not produce any significant protective effect in diabetogenesis produced by alloxan though it has hypoglycaemic effect.

Gamma radiation and its role in bio prosthetic aortic valves implanted in rat hearts

International Nuclear Information System (INIS)

Lamas, Gloria I.; Kairiyama, Eulogia; Navia, Jose

2000-01-01

Porcine heart valves glutaraldehyde fixed are implanted in patients with valvular deterioration. Mineralization may be the major factor in the long-term failure of tissue bio prosthesis. Gamma radiation randomly breaks some glutaraldehyde cross-links. As a consequence of irradiation, the polymeric fibers belonging to the valvular tissue are broken too, leading to sites of collagen fiber disorganisation. It is well known that the collagen fibers may act as a passive nucleator of salts where the calcium phosphate salts precipitate. This salt concentration has been described in association with disintegrated sites of protein fiber, which may favour new sites where the calcium salts would be deposit. The irradiation process is a technique used for sterilization of porcine heart valve. The main objective of this work was to study the effect of different doses of gamma radiation on the calcification process of subcutaneously implanted valves in rats. Small pieces from glutaraldehyde fixed valves, irradiated to different doses with a 60 Co sources were implanted subcutaneously in rats. The calcium was measured by X-ray and atomic absorption spectrophotometry. In our experimental conditions and at the radiation doses used in these tests, the calcium measurements on control and irradiated material were not significantly different. We conclude that, at the employed doses, the gamma radiation does not alter the process. (author) [es

Technical aspects of the integration of three-dimensional treatment planning dose parameters (GEC-ESTRO Working Group) into pre-implant planning for LDR gynecological interstitial brachytherapy.

Science.gov (United States)

Chi, A; Gao, M; Nguyen, N P; Albuquerque, K

2009-06-01

This study investigates the technical feasibility of pre-implant image-based treatment planning for LDR GYN interstitial brachytherapy(IB) based on the GEC-ESTRO guidelines. Initially, a virtual plan is generated based on the prescription dose and GEC-ESTRO defined OAR dose constraints with a pre-implant CT. After the actual implant, a regular diagnostic CT was obtained and fused with our pre-implant scan/initial treatment plan in our planning software. The Flexi-needle position changes, and treatment plan modifications were made if needed. Dose values were normalized to equivalent doses in 2 Gy fractions (LQED 2 Gy) derived from the linear-quadratic model with alpha/beta of 3 for late responding tissues and alpha/beta of 10 for early responding tissues. D(90) to the CTV, which was gross tumor (GTV) at the time of brachytherapy with a margin to count for microscopic disease, was 84.7 +/- 4.9% of the prescribed dose. The OAR doses were evaluated by D(2cc) (EBRT+IB). Mean D(2cc) values (LQED(2Gy)) for the rectum, bladder, sigmoid, and small bowel were the following: 63.7 +/- 8.4 Gy, 61.2 +/- 6.9 Gy, 48.0 +/- 3.5 Gy, and 49.9 +/- 4.2 Gy. This study confirms the feasibility of applying the GEC-ESTRO recommended dose parameters in pre-implant CT-based treatment planning in GYN IB. In the process, this pre-implant technique also demonstrates a good approximation of the target volume dose coverage, and doses to the OARs.

Endogenous lysophosphatidic acid participates in vascularisation and decidualisation at the maternal-fetal interface in the rat.

Science.gov (United States)

Sordelli, Micaela S; Beltrame, Jimena S; Zotta, Elsa; Gomez, Natalia; Dmytrenko, Ganna; Sales, María Elena; Blois, Sandra M; Davio, Carlos; Martinez, Silvina Perez; Franchi, Ana M; Ribeiro, María L

2017-10-01

Lysophosphatidic acid (LPA) affects several female reproductive functions through G-protein-coupled receptors. LPA contributes to embryo implantation via the lysophospholipid LPA 3 receptor. In the present study we investigated the participation of endogenous LPA signalling through the LPA 3 receptor in vascularisation and decidualisation, two crucial events at the maternal-fetal interface. Pregnant rats were treated with diacylglycerol pyrophosphate (DGPP), a highly selective antagonist of LPA 3 receptors, on Day 5 of gestation. Pregnant rats received intrauterine (i.u.) injections of single doses of DGPP (0.1mgkg -1 ) in a total volume of 2μL in the left horn (treated horn) in the morning of GD5. DGPP treatment produced aberrant embryo spacing and increased embryo resorption. The LPA 3 receptor antagonist decreased the cross-sectional length of the uterine and arcuate arteries and induced histological anomalies in the decidua and placentas. Marked haemorrhagic processes, infiltration of immune cells and tissue disorganisation were observed in decidual and placental tissues from sites of resorption. The mRNA expression of three vascularisation markers, namely interleukin 10 (Il10), vascular endothelial growth factor (Vegfa) and vascular endothelial growth factor receptor 1 (Vegfr1), was reduced at sites of resorption from Day 8. The results show that the disruption of endogenous LPA signalling by blocking the LPA 3 receptor modified the development of uterine vessels with consequences in the formation of the decidua and placenta and in the growth of embryos.

Effect of MeV energy He and N pre-implantation on the formation of porous silicon

Energy Technology Data Exchange (ETDEWEB)

Manuaba, A. E-mail: manu@rmki.kfki.hu; Paszti, F.; Ortega, C.; Grosman, A.; Horvath, Z.E.; Szilagyi, E.; Khanh, N.Q.; Vickridge, I

2001-06-01

The effects of MeV energy He and N pre-implantation of Si substrate on the structure of porous silicon formed by anodic etching were studied by measuring the depth profiles of {sup 15}N decorating the pores walls. Radiation damage was recovered by annealing after the implantation. It was found that the He implant accelerates the etching process, probably due to the bubbles or the remaining lattice damage. At a dose of 8x10{sup 16} ions/cm{sup 2} the He containing layer was formed with a significantly enhanced porosity due to the contribution of the large-sized bubbles. At the highest dose of 32.5x10{sup 16} ions/cm{sup 2} flaking took place during the anodic etching. In contrast to He, N stopped the anodic etching at a depth of critical N concentration of {approx}0.9 at.%. For the lowest implantation dose, where the peak concentration was below this limit, the pores propagate through the implanted layer with an enhanced speed.

The synergistic effect of TiO2 nanoporous modification and platelet-rich plasma treatment on titanium-implant stability in ovariectomized rats

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Jiang N

2016-09-01

Full Text Available Nan Jiang,1,2 Pinggong Du,2 Weidong Qu,2 Lin Li,2 Zhonghao Liu,2 Songsong Zhu1 1State Key Laboratory of Oral Diseases and Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu, 2Yantai City Stomatological Hospital, Yantai, People’s Republic of China Abstract: For several decades, titanium and its alloys have been commonly utilized for endosseous implantable materials, because of their good mechanical properties, chemical resistance, and biocompatibility. But associated low bone mass, wear and loss characteristics, and high coefficients of friction have limited their long-term stable performance, especially in certain abnormal bone-metabolism conditions, such as postmenopausal osteoporosis. In this study, we investigated the effects of platelet-rich plasma (PRP treatment and TiO2 nanoporous modification on the stability of titanium implants in osteoporotic bone. After surface morphology, topographical structure, and chemical changes of implant surface had been detected by scanning electron microscopy (SEM, atomic force microscopy, contact-angle measurement, and X-ray diffraction, we firstly assessed in vivo the effect of PRP treatment on osseointegration of TiO2-modified implants in ovariectomized rats by microcomputed tomography examinations, histology, biomechanical testing, and SEM observation. Meanwhile, the potential molecular mechanism involved in peri-implant osseous enhancement was also determined by quantitative real-time polymerase chain reaction. The results showed that this TiO2-modified surface was able to lead to improve bone implant contact, while PRP treatment was able to increase the implant surrounding bone mass. The synergistic effect of both was able to enhance the terminal force of implants drastically in biomechanical testing. Compared with surface modification, PRP treatment promoted earlier osteogenesis with increased expression of the RUNX2 and COL1 genes and

Embryotoxicity of benzalkonium chloride in vaginally treated rats.

Science.gov (United States)

Buttar, H S

1985-12-01

The effects of the spermicide benzalkonium chloride (BKC) were studied on the conceptus of rat. Single doses (0, 25, 50, 100 or 200 mg kg-1) of aqueous solutions of BKC were administered intravaginally (1 ml kg-1) on gestational day 1. The vulval metallic clips, used to prevent leakage of the solution, were removed 24 h post-treatment. Fetuses were obtained and examined for malformations on day 21 of gestation. slight to copious amounts of vaginal discharge and vaginitis were noticed in rats treated with the two largest doses of BKC. A dose-related increase in resorptions and fetal death, reduction in litter size and weight were observed in BKC-treated dams. The conceptus loss seemed to occur both before and after implantation. BKC did not cause any discernible visceral malformations, although minor sternal defects occurred in fetuses exposed to 100 and 200 mg kg-1 of the spermicide. These results suggest that single vaginal application of BKC is embryo- and fetocidal in the rat at a dose about 143 times higher than that recommended for controlling conception in women.

Evaluation of Musa (Paradisiaca Linn. cultivar)--"Puttubale" stem juice for antilithiatic activity in albino rats.

Science.gov (United States)

Prasad, K V; Bharathi, K; Srinivasan, K K

1993-10-01

The fresh juice of Musa stem (Puttubale) was tested for its antilithiatic activity. Zinc discs were implanted in the urinary bladder of albino rats to induce urolithiasis. The stones formed were mainly of magnesium ammonium phosphate with traces of calcium oxalate. Musa stem juice (3 mL/rat/day orally) was found to be effective in reducing the formation and also in dissolving the pre-formed stones.

A comparative study of zirconium and titanium implants in rat: osseointegration and bone material quality.

Science.gov (United States)

Hoerth, Rebecca M; Katunar, María R; Gomez Sanchez, Andrea; Orellano, Juan C; Ceré, Silvia M; Wagermaier, Wolfgang; Ballarre, Josefina

2014-02-01

Permanent metal implants are widely used in human medical treatments and orthopedics, for example as hip joint replacements. They are commonly made of titanium alloys and beyond the optimization of this established material, it is also essential to explore alternative implant materials in view of improved osseointegration. The aim of our study was to characterize the implant performance of zirconium in comparison to titanium implants. Zirconium implants have been characterized in a previous study concerning material properties and surface characteristics in vitro, such as oxide layer thickness and surface roughness. In the present study, we compare bone material quality around zirconium and titanium implants in terms of osseointegration and therefore characterized bone material properties in a rat model using a multi-method approach. We used light and electron microscopy, micro Raman spectroscopy, micro X-ray fluorescence and X-ray scattering techniques to investigate the osseointegration in terms of compositional and structural properties of the newly formed bone. Regarding the mineralization level, the mineral composition, and the alignment and order of the mineral particles, our results show that the maturity of the newly formed bone after 8 weeks of implantation is already very high. In conclusion, the bone material quality obtained for zirconium implants is at least as good as for titanium. It seems that the zirconium implants can be a good candidate for using as permanent metal prosthesis for orthopedic treatments.

Comparative study of as-implanted and pre-damaged ion-beam-synthesized ZnS nanocrystallites in SiO sub 2

CERN Document Server

Gao, K Y; Grosshans, I; Hipp, W; Stritzker, B

2002-01-01

The semiconducting ZnS nanocrystallites were synthesized by sequential high dose ion implantation of Zn and S in thermally grown SiO sub 2 on Si(1 0 0) and subsequent rapid thermal annealing (RTA). Some samples were pre-implanted with Ar ions in order to investigate the influence of radiation induced damage on the formation of ZnS nanocrystallites. The crystal structure of the ZnS crystallites, their size distribution and the concentration depth profile were analyzed by X-ray diffraction (XRD), Rutherford backscattering spectroscopy (RBS) and cross-sectional transmission-electron-microscopy (XTEM). The XRD results indicate, that the phase transition from cubic zinc blende to hexagonal wurtzite structure of ZnS nanocrystallites begins at temperatures below 1000 degree sign C. The RBS results show a clear redistribution of Zn and S after RTA annealing. The concentration of Zn is seriously reduced due to strong diffusion towards deeper regions and the surface, while Ar pre-implantation partially suppressed the c...

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

African Journals Online (AJOL)

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

Automated microinjection of recombinant BCL-X into mouse zygotes enhances embryo development.

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Xinyu Liu

Full Text Available Progression of fertilized mammalian oocytes through cleavage, blastocyst formation and implantation depends on successful implementation of the developmental program, which becomes established during oogenesis. The identification of ooplasmic factors, which are responsible for successful embryo development, is thus crucial in designing possible molecular therapies for infertility intervention. However, systematic evaluation of molecular targets has been hampered by the lack of techniques for efficient delivery of molecules into embryos. We have developed an automated robotic microinjection system for delivering cell impermeable compounds into preimplantation embryos with a high post-injection survival rate. In this paper, we report the performance of the system on microinjection of mouse embryos. Furthermore, using this system we provide the first evidence that recombinant BCL-XL (recBCL-XL protein is effective in preventing early embryo arrest imposed by suboptimal culture environment. We demonstrate that microinjection of recBCL-XL protein into early-stage embryos repairs mitochondrial bioenergetics, prevents reactive oxygen species (ROS accumulation, and enhances preimplantation embryo development. This approach may lead to a possible treatment option for patients with repeated in vitro fertilization (IVF failure due to poor embryo quality.

Maintained functionality of an implantable radiotelemetric blood pressure and heart rate sensor after magnetic resonance imaging in rats

International Nuclear Information System (INIS)

Nölte, I; Boll, H; Figueiredo, G; Groden, C; Brockmann, M A; Gorbey, S; Lemmer, B

2011-01-01

Radiotelemetric sensors for in vivo assessment of blood pressure and heart rate are widely used in animal research. MRI with implanted sensors is regarded as contraindicated as transmitter malfunction and injury of the animal may be caused. Moreover, artefacts are expected to compromise image evaluation. In vitro, the function of a radiotelemetric sensor (TA11PA-C10, Data Sciences International) after exposure to MRI up to 9.4 T was assessed. The magnetic force of the electromagnetic field on the sensor as well as radiofrequency (RF)-induced sensor heating was analysed. Finally, MRI with an implanted sensor was performed in a rat. Imaging artefacts were analysed at 3.0 and 9.4 T ex vivo and in vivo. Transmitted 24 h blood pressure and heart rate were compared before and after MRI to verify the integrity of the telemetric sensor. The function of the sensor was not altered by MRI up to 9.4 T. The maximum force exerted on the sensor was 273 ± 50 mN. RF-induced heating was ruled out. Artefacts impeded the assessment of the abdomen and thorax in a dead rat, but not of the head and neck. MRI with implanted radiotelemetric sensors is feasible in principal. The tested sensor maintains functionality up to 9.4 T. Artefacts hampered abdominal and throacic imaging in rats, while assessment of the head and neck is possible

Assessment of implantable infusion pumps for continuous infusion of human insulin in rats: potential for group housing

DEFF Research Database (Denmark)

Jensen, Vivi Flou Hjorth; Molck, Anne-Marie; Martensson, Martin

2017-01-01

compound in these studies, and a comparator model of persistent exposure by HI infusion from external pumps has recently been developed to support toxicological evaluation of long-acting insulin analogues. However, this model requires single housing of the animals. Developing an insulin-infusion model...... which allows group housing would therefore greatly improve animal welfare. The aim of the present study was to investigate the suitability of implantable infusion pumps for HI infusion in group-housed rats. Group housing of rats implanted with a battery-driven pump proved to be possible. Intravenous...... infusion of HI lowered blood glucose levels persistently for two weeks, providing a comparator model for use in two-week repeated-dose toxicity studies with new long-acting insulin analogues, which allows group housing, and thereby increasing animal welfare compared with an external infusion model....

Effect of ovariectomy and 17 beta-estradiol implantation on bone metabolism in female rats fed 1,25 dihydroxyvitamin D3

International Nuclear Information System (INIS)

Osborne, M.; Sherman, S.; Soares, J.H. Jr.

1986-01-01

Eight-week old Sprague-Dawley female rats were used in two experiments to test the effects of 17-B estradiol (E 2 ) via silastic tubing implants on 3 H-labelled tetracycline (TC) incorporation into bone. 1,25(OH) 2 D 3 was the dietary source of vitamin D in a low calcium (.2%) semipurified eggwhite diet. In experiment I, the Ovx + E 2 animals showed a significantly (p 3 H TC uptake in scapula during a 2-week labelling experiment. An increase in 3 H TC content resulted (p 2 was implanted in 1,25(OH) 2 D 3 fed Ovx rats. However, the calcium content was not significantly different. The effect of dietary 1,25(OH) 2 D 3 and E 2 implantation appears to be additive. Optimal action of the Vitamin D endocrine system may be dependent on presence of E 2

Challenges of animal models in SCI research: Effects of pre-injury task-specific training in adult rats before lesion.

Science.gov (United States)

May, Zacnicte; Fouad, Karim; Shum-Siu, Alice; Magnuson, David S K

2015-09-15

A rarely explored subject in animal research is the effect of pre-injury variables on behavioral outcome post-SCI. Low reporting of such variables may underlie some discrepancies in findings between laboratories. Particularly, intensive task-specific training before a SCI might be important, considering that sports injuries are one of the leading causes of SCI. Thus, individuals with SCI often underwent rigorous training before their injuries. In the present study, we asked whether training before SCI on a grasping task or a swimming task would influence motor recovery in rats. Swim pre-training impaired recovery of swimming 2 and 4 weeks post-injury. This result fits with the idea of motor learning interference, which posits that learning something new may disrupt learning of a new task; in this case, learning strategies to compensate for functional loss after SCI. In contrast to swimming, grasp pre-training did not influence grasping ability after SCI at any time point. However, grasp pre-trained rats attempted to grasp more times than untrained rats in the first 4 weeks post-injury. Also, lesion volume of grasp pre-trained rats was greater than that of untrained rats, a finding which may be related to stress or activity. The increased participation in rehabilitative training of the pre-trained rats in the early weeks post-injury may have potentiated spontaneous plasticity in the spinal cord and counteracted the deleterious effect of interference and bigger lesions. Thus, our findings suggest that pre-training plays a significant role in recovery after CNS damage and needs to be carefully controlled for. Copyright © 2015 Elsevier B.V. All rights reserved.

Pre-exposure Schedule Effects on Generalization of Taste Aversion and Palatability for Thirsty and Not Thirsty Rats

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Rocío Angulo

2018-06-01

Full Text Available The study reported four experiments aiming to test the effects of the pre-exposure schedule and water deprivation on the generalization of a conditioned taste aversion in rats, with a particular focus on testing whether or not the concurrent schedule might enhance generalization. In two experiments, non-water-deprived rats received concurrent, intermixed, or blocked exposure to a sweet-acid solution and a salty-acid solution before conditioning of one of these compounds and testing of both flavors. During pre-exposure, the rats consumed a greater amount of the sweet-acid solution than the salty-acid solution (Experiments 1 and 2, consumption of the former increasing during pre-exposure while consumption of the latter decreased (Experiment 1. Furthermore, consumption of the salty-acid solution was lower during concurrent than intermixed or blocked pre-exposure (Experiment 1 and 2 while consumption of the sweet-acid solution was greater during intermixed than concurrent or blocked pre-exposure (Experiment 1. It is discussed whether the pre-exposure schedule might modify stimulus perception beyond the mere enhancement of stimulus differentiation, by, for instance, affecting the palatability of gustatory stimuli. Evidence for enhanced generalization after concurrent pre-exposure was not found for either deprived (Experiments 1, 2, and 3 or non-deprived rats (Experiments 3 and 4, with deprivation leading to a general increase in consumption of both the conditioned and test flavors. This then raised the question of whether or not concurrent pre-exposure to flavors always increases generalization between them. The present study highlights the importance of this issue for various accounts of perceptual learning.

The effects of metformin on ovum implantation and pregnancy outcome in rats with induced PCOS

Directory of Open Access Journals (Sweden)

Mesbah F

2011-06-01

Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Polycystic ovary syndrome (PCOS is the most common cause of anovulatory infertility. Metformin which is effectively used for the treatment of anovulatory PCOS improves pregnancy rate and endometrial receptivity and reduces the risk of miscarriage. The aim of this study was to evaluate the effects of metformin on the endometrium, the number of fetuses and hormonal levels of PCOS rats."n"nMethods: Forty female adult Sprague-Dawley rats were assigned randomly into four equal groups. Group I: control rats, group II: rats receiving metformin (150 mg/kg/day, group III: Estradiol Valerate-induced PCOS rats (4 mg/rat and group IV: induced PCOS rats receiving metformin. Body weight and serum levels of glucose, LH, FSH, testosterone, progesterone and estradiol were measured. Following mating, each group was divided into two subgroups and the rats were sacrificed on the 5th and 15th day of gestation to evaluate endometrial reaction to implantation and fetus count, respectively."n"nResults: Hormone assay showed a significant increase in testosterone, estradiol, LH, FSH and blood glucose levels in group III compared to the controls (P≤0.01 and a significant decrease in blood glucose in group IV versus group III (P≤0

Lithium chloride enhances bone regeneration and implant osseointegration in osteoporotic conditions.

Science.gov (United States)

Jin, Yifan; Xu, Lihua; Hu, Xiaohui; Liao, Shixian; Pathak, Janak L; Liu, Jinsong

2016-10-06

Osteoporotic patients have a high risk of dental and orthopedic implant failure. Lithium chloride (LiCl) has been reported to enhance bone formation. However, the role of LiCl in the success rate of dental and orthopedic implants in osteoporotic conditions is still unknown. We investigated whether LiCl enhances implant osseointegration, implant fixation, and bone formation in osteoporotic conditions. Sprague-Dawley female rats (n = 18) were ovariectomized (OVX) to induce osteoporosis, and another nine rats underwent sham surgery. Three months after surgery, titanium implants were implanted in the tibia of the OVX and sham group rats. After implantation, the OVX rats were gavaged with 150 mg/kg/2 days of LiCl (OVX + LiCl group) or saline (OVX group), and sham group rats were gavaged with saline for 3 months. Implant osseointegration and bone formation were analyzed using histology, biomechanical testing, and micro computed tomography (micro-CT). More bone loss was observed in the OVX group compared to the control, and LiCl treatment enhanced bone formation and implant fixation in osteoporotic rats. In the OVX group, bone-implant contact (BIC) was decreased by 81.2 % compared to the sham group. Interestingly, the OVX + LiCl group showed 4.4-fold higher BIC compared to the OVX group. Micro-CT data of tibia from the OVX + LiCl group showed higher bone volume, trabecular thickness, trabecular number, and osseointegration compared to the OVX group. Maximum push-out force and implant-bone interface shear strength were 2.9-fold stronger in the OVX + LiCl group compared to the OVX group. In conclusion, LiCl enhanced implant osseointegration, implant fixation, and bone formation in osteoporotic conditions, suggesting LiCl as a promising therapeutic agent to prevent implant failure and bone loss in osteoporotic conditions.

High-grade Angiosarcoma Associated with Ruptured Breast Implants

Directory of Open Access Journals (Sweden)

Nicolas R. Smoll, MBBS

2013-04-01

Full Text Available Summary: Since the serendipitous discovery that implanted polymers cause sarcomas in rats, much research has been conducted to prove or disprove a link between silicone breast implants and/or polymer-based materials and breast cancer. In light of an initial report that 35% of rats implanted with a variety of polymers developed fibrosarcomas, we report a case of primary angiosarcoma found in a patient presenting with bilateral rupture of gel-filled breast implants.

The effect of x-irradiation on the implantation and development of the white rat fetus

International Nuclear Information System (INIS)

Danius, J.; Bahauddin, R.

1976-01-01

X-ray whole body irradiation (200R, exposure dose) was performed on young virgin females of about 4 months old. The female rats, divided into three groups, were treated as follows: Female rats as control (unirradiated = 1Kt): female rats irradiated before mating (RSbK), and female rats irradiated after mating (RSsK). The average number of surviving foetuses at 20 days gestational stage of 1Kt was compared with that of RSbK, and no significant difference was found (P<=0.05) while the difference between the average number of RSsK foetuses compared with that of 1Kt and RSbK was highly significant (P<0.01). A decrease in the average number of surviving foetuses was found in RSsK, although the analysis of variance of all groups, revealed no significant difference (P<=0.05) in the average number of implantations and weight of the foetuses. (author)

Silicon-Doped Titanium Dioxide Nanotubes Promoted Bone Formation on Titanium Implants.

Science.gov (United States)

Zhao, Xijiang; Wang, Tao; Qian, Shi; Liu, Xuanyong; Sun, Junying; Li, Bin

2016-02-26

While titanium (Ti) implants have been extensively used in orthopaedic and dental applications, the intrinsic bioinertness of untreated Ti surface usually results in insufficient osseointegration irrespective of the excellent biocompatibility and mechanical properties of it. In this study, we prepared surface modified Ti substrates in which silicon (Si) was doped into the titanium dioxide (TiO₂) nanotubes on Ti surface using plasma immersion ion implantation (PIII) technology. Compared to TiO₂ nanotubes and Ti alone, Si-doped TiO₂ nanotubes significantly enhanced the expression of genes related to osteogenic differentiation, including Col-I, ALP, Runx2, OCN, and OPN, in mouse pre-osteoblastic MC3T3-E1 cells and deposition of mineral matrix. In vivo, the pull-out mechanical tests after two weeks of implantation in rat femur showed that Si-doped TiO₂ nanotubes improved implant fixation strength by 18% and 54% compared to TiO₂-NT and Ti implants, respectively. Together, findings from this study indicate that Si-doped TiO₂ nanotubes promoted the osteogenic differentiation of osteoblastic cells and improved bone-Ti integration. Therefore, they may have considerable potential for the bioactive surface modification of Ti implants.

Bone tissue engineering for spine fusion : An experimental study on ectopic and orthotopic implants in rats

NARCIS (Netherlands)

van Gaalen, SM; Dhert, WJA; van den Muysenberg, A; Oner, FC; van Blitterswijk, C; Verbout, AJ; de Bruijn, J.D.

2004-01-01

Alternatives to the use of autologous bone as a bone graft in spine surgery are needed. The purpose of this study was to examine tissue-engineered bone constructs in comparison with control scaffolds without cells in a posterior spinal implantation model in rats. Syngeneic bone marrow cells were

Therapeutic interaction of systemically-administered mesenchymal stem cells with peri-implant mucosa.

Directory of Open Access Journals (Sweden)

Ryosuke Kondo

Full Text Available OBJECTIVES: The objective of this study was to investigate the effect of systemically transplanted mesenchymal stem cells (MSCs on the peri-implant epithelial sealing around dental implants. MATERIALS AND METHODS: MSCs were isolated from bone marrow of donor rats and expanded in culture. After recipient rats received experimental titanium dental implants in the bone sockets after extraction of maxillary right first molars, donor rat MSCs were intravenously transplanted into the recipient rats. RESULTS: The injected MSCs were found in the oral mucosa surrounding the dental implants at 24 hours post-transplantation. MSC transplantation accelerated the formation of the peri-implant epithelium (PIE-mediated mucosa sealing around the implants at an early stage after implantation. Subsequently, enhanced deposition of laminin-332 was found along the PIE-implant interface at 4 weeks after the replacement. We also observed enhanced attachment and proliferation of oral mucous epithelial cells. CONCLUSION: Systemically transplanted MSCs might play a critical role in reinforcing the epithelial sealing around dental implants.

The effect of Ni pre-implantation on surface morphology and optical absorption properties of Ag nanoparticles embedded in SiO2

International Nuclear Information System (INIS)

Shen, Yanyan; Qi, Ting; Qiao, Yu; Yu, Shengwang; Hei, Hongjun; He, Zhiyong

2016-01-01

Graphical abstract: - Highlights: • Ag concentration increased significantly due to the Ni pre-implantation. • Deposition and accumulation process of Ag atoms depends on Ni fluences. • The incorporation of Ni elements in Ag NPs can damp SPR absorption intensity. • AgNi alloy NPs embedded in SiO 2 have been created by sequentially implantation. • Unique SPR absorption with dual peaks centered at 406 nm and 563 nm was observed. - Abstract: The effect of Ni ion fluence on Ag nucleation and particle growth was investigated by sequentially implantation of 60 keV Ni ions at fluences of 1 × 10 16 , 5 × 10 16 , 1 × 10 17 ions/cm 2 and 70 keV Ag ions at a fluence of 5 × 10 16 ions/cm 2 . Due to the modification of the deposition and accumulation process of Ag implants caused by Ni pre-implantation, the surface morphology, structures, and optical absorption properties of the Ag nanoparticles (NPs) depends strongly on the Ni fluences. UV–vis absorption spectroscopy study showed that the introducing of Ni atoms lead to intensity decrease in the Ag SPR band. Remarkable local concentration increase of Ag profiles appeared for the sample pre-implanted by Ni ions of 5.0 × 10 16 ions/cm 2 . In particular, the AgNi alloy NPs with dual absorption peaks centered at 406 nm and 563 nm have been formed after 600 °C annealing in Ar atmosphere. However, at a low fluence of 1.0 × 10 16 ions/cm 2 , only small increase of the local Ag concentration than the Ag ions singly implanted sample can be observed. At a high fluence of 1.0 × 10 17 ions/cm 2 , lots Ag atoms are trapped close to the surface, which result in heavy sputtering loss of Ag atoms and the sublimation of Ag atoms after 600 °C annealing.

Birth of rats following nuclear exchange at the 2-cell stage.

Science.gov (United States)

Roh, Sangho; Guo, Jitong; Malakooti, Nakisa; Morrison, John R; Trounson, Alan O; Du, Zhong Tao

2003-11-01

We report full-term development of nuclear transfer embryos following nuclear exchange at the 2-cell stage. Nuclei from 2-cell rat embryos were transferred into enucleated 2-cell embryos and developed to term after transfer to recipients (NT2). Pronuclear exchange in zygotes was used for comparison (NT1). Zygotes and 2-cell embryos were harvested from 4-week-old female Sprague-Dawley rats. Nuclear transfer was performed by transferring the pronuclei or karyoplasts into the perivitelline space of recipient embryos followed by electrofusion to reconstruct embryos. Fused couplets were cultured for 4 or 24 h before being transferred into day 1 pseudopregnant recipients (Hooded Wistar) at the 1- or 2-cell stage. In vitro culture was also carried out to check the developmental competence of the embryos. In vitro development to the blastocyst stage was not significantly different between the two groups (NT1, 34.3%; NT2, 45.0%). Two of three recipients from NT1 and two of five recipients from NT2 became pregnant. Six pups (3 from NT1, 3 from NT2) were delivered from the four foster mothers. Three female pups survived; 2 from NT1 and 1 from NT2. At 2 months of age these pups appeared healthy, and were mated with Sprague-Dawley males. One rat derived from NT1 delivered 15 pups (5 males, 10 females) as did the rat from NT2 (7 males, 8 females). Our results show that by using karyoplasts from 2-cell stage embryos as nuclear donors and reconstructing them with enucleated 2-cell embryos, healthy rats can be produced.

Curcumin Implants, not Curcumin Diet Inhibits Estrogen-Induced Mammary Carcinogenesis in ACI Rats

Science.gov (United States)

Bansal, Shyam S.; kausar, Hina; Vadhanam, Manicka V.; Ravoori, Srivani; Pan, Jianmin; Rai, Shesh N.; Gupta, Ramesh C.

2014-01-01

Curcumin is widely known for its anti-oxidant, anti-inflammatory and anti-proliferative activities in cell culture studies. However, poor oral bioavailability limited its efficacy in animal and clinical studies. Recently, we developed polymeric curcumin implants that circumvents oral bioavailability issues, and tested their potential against 17β-estradiol (E2)-mediated mammary tumorigenesis. Female ACI rats were administered curcumin either via diet (1,000 ppm) or via polymeric curcumin implants (two 2-cm; 200 mg each; 20% drug load) 4 days prior to grafting a subcutaneous E2 silastic implant (1.2 cm, 9 mg E2). Implants were changed after 4½ months to provide higher curcumin dose at the appearance of palpable tumors. The animals were euthanized after 3 weeks, 3 months and after the tumor incidence reached >80% (~6 months) in control animals. The curcumin administered via implants resulted in significant reduction in both the tumor multiplicity (2±1 vs 5±3; p=0.001) and tumor volume (184±198 mm3 vs 280±141 mm3; p=0.0283); the dietary curcumin, however, was ineffective. Dietary curcumin increased hepatic CYP1A and CYP1B1 activities without any effect on CYP3A4 activity whereas curcumin implants increased both CYP1A and CYP3A4 activities but decreased CYP1B1 activity in presence of E2. Since CYP1A and 3A4 metabolize most of the E2 to its non-carcinogenic 2-OH metabolite and CYP1B1 produces potentially carcinogenic 4-OH metabolite, favorable modulation of these CYPs via systemically delivered curcumin could be one of the potential mechanisms. The analysis of plasma and liver by HPLC showed substantially higher curcumin levels via implants versus the dietary route despite substantially higher dose administered. PMID:24501322

Small-Group Phonological Awareness Training for Pre-Kindergarten Children with Hearing Loss Who Wear Cochlear Implants and/or Hearing Aids

Science.gov (United States)

Werfel, Krystal L.; Douglas, Michael; Ackal, Leigh

2016-01-01

This case report details a year-long phonological awareness (PA) intervention for pre-kindergarten children with hearing loss (CHL) who use listening and spoken language. All children wore cochlear implants and/or hearing aids. Intervention occurred for 15 min/day, 4 days per week across the pre-kindergarten school year and was delivered by…

Human embryos secrete microRNAs into culture media--a potential biomarker for implantation.

Science.gov (United States)

Rosenbluth, Evan M; Shelton, Dawne N; Wells, Lindsay M; Sparks, Amy E T; Van Voorhis, Bradley J

2014-05-01

To determine whether human blastocysts secrete microRNA (miRNAs) into culture media and whether these reflect embryonic ploidy status and can predict in vitro fertilization (IVF) outcomes. Experimental study of human embryos and IVF culture media. Academic IVF program. 91 donated, cryopreserved embryos that developed into 28 tested blastocysts, from 13 couples who had previously completed IVF cycles. None. Relative miRNA expression in IVF culture media. Blastocysts were assessed by chromosomal comparative genomic hybridization analysis, and the culture media from 55 single-embryo transfer cycles was tested for miRNA expression using an array-based quantitative real-time polymerase chain reaction analysis. The expression of the identified miRNA was correlated with pregnancy outcomes. Ten miRNA were identified in the culture media; two were specific to spent media (miR-191 and miR-372), and one was only present in media before the embryos had been cultured (miR-645). MicroRNA-191 was more highly concentrated in media from aneuploid embryos, and miR-191, miR-372, and miR-645 were more highly concentrated in media from failed IVF/non-intracytoplasmic sperm injection cycles. Additionally, miRNA were found to be more highly concentrated in ICSI and day-5 media samples when compared with regularly inseminated and day-4 samples, respectively. MicroRNA can be detected in IVF culture media. Some of these miRNA are differentially expressed according to the fertilization method, chromosomal status, and pregnancy outcome, which makes them potential biomarkers for predicting IVF success. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Untwisting the Caenorhabditis elegans embryo

Science.gov (United States)

Christensen, Ryan Patrick; Bokinsky, Alexandra; Santella, Anthony; Wu, Yicong; Marquina-Solis, Javier; Guo, Min; Kovacevic, Ismar; Kumar, Abhishek; Winter, Peter W; Tashakkori, Nicole; McCreedy, Evan; Liu, Huafeng; McAuliffe, Matthew; Mohler, William; Colón-Ramos, Daniel A; Bao, Zhirong; Shroff, Hari

2015-01-01

The nematode Caenorhabditis elegans possesses a simple embryonic nervous system with few enough neurons that the growth of each cell could be followed to provide a systems-level view of development. However, studies of single cell development have largely been conducted in fixed or pre-twitching live embryos, because of technical difficulties associated with embryo movement in late embryogenesis. We present open-source untwisting and annotation software (http://mipav.cit.nih.gov/plugin_jws/mipav_worm_plugin.php) that allows the investigation of neurodevelopmental events in late embryogenesis and apply it to track the 3D positions of seam cell nuclei, neurons, and neurites in multiple elongating embryos. We also provide a tutorial describing how to use the software (Supplementary file 1) and a detailed description of the untwisting algorithm (Appendix). The detailed positional information we obtained enabled us to develop a composite model showing movement of these cells and neurites in an 'average' worm embryo. The untwisting and cell tracking capabilities of our method provide a foundation on which to catalog C. elegans neurodevelopment, allowing interrogation of developmental events in previously inaccessible periods of embryogenesis. DOI: http://dx.doi.org/10.7554/eLife.10070.001 PMID:26633880

Untwisting the Caenorhabditis elegans embryo.

Science.gov (United States)

Christensen, Ryan Patrick; Bokinsky, Alexandra; Santella, Anthony; Wu, Yicong; Marquina-Solis, Javier; Guo, Min; Kovacevic, Ismar; Kumar, Abhishek; Winter, Peter W; Tashakkori, Nicole; McCreedy, Evan; Liu, Huafeng; McAuliffe, Matthew; Mohler, William; Colón-Ramos, Daniel A; Bao, Zhirong; Shroff, Hari

2015-12-03

The nematode Caenorhabditis elegans possesses a simple embryonic nervous system with few enough neurons that the growth of each cell could be followed to provide a systems-level view of development. However, studies of single cell development have largely been conducted in fixed or pre-twitching live embryos, because of technical difficulties associated with embryo movement in late embryogenesis. We present open-source untwisting and annotation software (http://mipav.cit.nih.gov/plugin_jws/mipav_worm_plugin.php) that allows the investigation of neurodevelopmental events in late embryogenesis and apply it to track the 3D positions of seam cell nuclei, neurons, and neurites in multiple elongating embryos. We also provide a tutorial describing how to use the software (Supplementary file 1) and a detailed description of the untwisting algorithm (Appendix). The detailed positional information we obtained enabled us to develop a composite model showing movement of these cells and neurites in an 'average' worm embryo. The untwisting and cell tracking capabilities of our method provide a foundation on which to catalog C. elegans neurodevelopment, allowing interrogation of developmental events in previously inaccessible periods of embryogenesis.

Comparative studies on permanent prostate brachytherapy: pre-plan and real-time transrectal ultrasound guided iodine-125 seed implants at Korle-Bu Teaching Hospital, Ghana

International Nuclear Information System (INIS)

Kalolo, L.T.

2013-06-01

This research was carried out to investigate and compare the real-time and pre-plan implant at the Radiotherapy Department of the Korle Bu Teaching Hospital, Ghana. Prowess Panther 4.5 treatment planning system and variseed 7.2 software were used for pre-plan and real-time implant respectively. The study was conducted for eighty three (83) patients treated for prostate cancer through real-time implant brachytherapy between september, 2008 to April, 2013. Thirty one patients (31) patients whose ultrasound images were available were selected for the pre-plan study. The slices of ultrasound images were re-drawn on transparent A-4 sheets and later on scanned, contoured and registered in the treatment planning system (prowess 4.5). After planning, the volume to be implanted, total number of needles, seeds and the total activity of the source were displayed. Comparison was done withe the pre-plan and real-time implant. In both cases the variation was below 5% as recommended in dosimetry. About 30% - 40% of the imported seeds were left un-used due to over-estimation of seeds ordered from the manufacturer (BARD Company-USA). Hence this work (pre-plan) aims to solve this problem. The comparison for dosimetric parameters was assessed for prostate, urethra and rectum as (V 95%, V 100%, V 150%, D90Gy, D90%), (D90Gy, D90%, D30Gy, D30% ) and (V 100%, D30Gy and D30%) respectively and the variation were within the limit of ± 5%. Comparison of dosimetric values for this work were done with other institutions, like Karolinska university hospital, Sweden, The institute of Curie/ hospital Cochin Group Paris-France and European recommendations. The values reported at Korle - Bu teaching hospital (this work) were in good agreement with the international guidelines. (au)

Scaffold-Free Tubular Tissues Created by a Bio-3D Printer Undergo Remodeling and Endothelialization when Implanted in Rat Aortae

Science.gov (United States)

Itoh, Manabu; Nakayama, Koichi; Noguchi, Ryo; Kamohara, Keiji; Furukawa, Kojirou; Uchihashi, Kazuyoshi; Toda, Shuji; Oyama, Jun-ichi; Node, Koichi; Morita, Shigeki

2015-01-01

Background Small caliber vascular prostheses are not clinically available because synthetic vascular prostheses lack endothelial cells which modulate platelet activation, leukocyte adhesion, thrombosis, and the regulation of vasomotor tone by the production of vasoactive substances. We developed a novel method to create scaffold-free tubular tissue from multicellular spheroids (MCS) using a “Bio-3D printer”-based system. This system enables the creation of pre-designed three-dimensional structures using a computer controlled robotics system. With this system, we created a tubular structure and studied its biological features. Methods and Results Using a “Bio-3D printer,” we made scaffold-free tubular tissues (inner diameter of 1.5 mm) from a total of 500 MCSs (2.5× 104 cells per one MCS) composed of human umbilical vein endothelial cells (40%), human aortic smooth muscle cells (10%), and normal human dermal fibroblasts (50%). The tubular tissues were cultured in a perfusion system and implanted into the abdominal aortas of F344 nude rats. We assessed the flow by ultrasonography and performed histological examinations on the second (n = 5) and fifth (n = 5) day after implantation. All grafts were patent and remodeling of the tubular tissues (enlargement of the lumen area and thinning of the wall) was observed. A layer of endothelial cells was confirmed five days after implantation. Conclusions The scaffold-free tubular tissues made of MCS using a Bio-3D printer underwent remodeling and endothelialization. Further studies are warranted to elucidate the underlying mechanism of endothelialization and its function, as well as the long-term results. PMID:26325298

Scaffold-Free Tubular Tissues Created by a Bio-3D Printer Undergo Remodeling and Endothelialization when Implanted in Rat Aortae.

Science.gov (United States)

Itoh, Manabu; Nakayama, Koichi; Noguchi, Ryo; Kamohara, Keiji; Furukawa, Kojirou; Uchihashi, Kazuyoshi; Toda, Shuji; Oyama, Jun-Ichi; Node, Koichi; Morita, Shigeki

2015-01-01

Small caliber vascular prostheses are not clinically available because synthetic vascular prostheses lack endothelial cells which modulate platelet activation, leukocyte adhesion, thrombosis, and the regulation of vasomotor tone by the production of vasoactive substances. We developed a novel method to create scaffold-free tubular tissue from multicellular spheroids (MCS) using a "Bio-3D printer"-based system. This system enables the creation of pre-designed three-dimensional structures using a computer controlled robotics system. With this system, we created a tubular structure and studied its biological features. Using a "Bio-3D printer," we made scaffold-free tubular tissues (inner diameter of 1.5 mm) from a total of 500 MCSs (2.5× 104 cells per one MCS) composed of human umbilical vein endothelial cells (40%), human aortic smooth muscle cells (10%), and normal human dermal fibroblasts (50%). The tubular tissues were cultured in a perfusion system and implanted into the abdominal aortas of F344 nude rats. We assessed the flow by ultrasonography and performed histological examinations on the second (n = 5) and fifth (n = 5) day after implantation. All grafts were patent and remodeling of the tubular tissues (enlargement of the lumen area and thinning of the wall) was observed. A layer of endothelial cells was confirmed five days after implantation. The scaffold-free tubular tissues made of MCS using a Bio-3D printer underwent remodeling and endothelialization. Further studies are warranted to elucidate the underlying mechanism of endothelialization and its function, as well as the long-term results.

Long-term neuroplasticity of the face primary motor cortex and adjacent somatosensory cortex induced by tooth loss can be reversed following dental implant replacement in rats.

Science.gov (United States)

Avivi-Arber, Limor; Lee, Jye-Chang; Sood, Mandeep; Lakschevitz, Flavia; Fung, Michelle; Barashi-Gozal, Maayan; Glogauer, Michael; Sessle, Barry J

2015-11-01

Tooth loss is common, and exploring the neuroplastic capacity of the face primary motor cortex (face-M1) and adjacent primary somatosensory cortex (face-S1) is crucial for understanding how subjects adapt to tooth loss and their prosthetic replacement. The aim was to test if functional reorganization of jaw and tongue motor representations in the rat face-M1 and face-S1 occurs following tooth extraction, and if subsequent dental implant placement can reverse this neuroplasticity. Rats (n = 22) had the right maxillary molar teeth extracted under local and general anesthesia. One month later, seven rats had dental implant placement into healed extraction sites. Naive rats (n = 8) received no surgical treatment. Intracortical microstimulation (ICMS) and recording of evoked jaw and tongue electromyographic responses were used to define jaw and tongue motor representations at 1 month (n = 8) or 2 months (n = 7) postextraction, 1 month postimplant placement, and at 1-2 months in naive rats. There were no significant differences across study groups in the onset latencies of the ICMS-evoked responses (P > 0.05), but in comparison with naive rats, tooth extraction caused a significant (P rats. These novel findings suggest that face-M1 and adjacent face-S1 may play a role in adaptive mechanisms related to tooth loss and their replacement with dental implants. © 2015 Wiley Periodicals, Inc.

The impact of food intake and social habits on embryo quality and the likelihood of blastocyst formation.

Science.gov (United States)

Braga, Daniela Paes Almeida Ferreira; Halpern, Gabriela; Setti, Amanda S; Figueira, Rita Cássia S; Iaconelli, Assumpto; Borges, Edson

2015-07-01

The aim of this study was to evaluate the influence of patients' lifestyle factors and eating habits on embryo development. A total of 2659 embryos recovered from 269 patients undergoing intracytoplasmic sperm injection cycles were included. The frequency of intake of food items and social habits were registered and its influences on embryo development evaluated. The consumption of cereals, vegetables and fruits positively influenced the embryo quality at the cleavage stage. The quality of the embryo at the cleavage stage was also negatively correlated with the consumption of alcoholic drinks and smoking habits. The consumption of fruits influenced the likelihood of blastocyst formation, which was also positively affected by the consumption of fish. Being on a weight-loss diet and consumption of red meat had a negative influence on the likelihood of blastocyst formation. The likelihood of blastocyst formation was also negatively influenced by the consumption of alcoholic drinks and by smoking habits. The consumption of red meat and body mass index had a negative effect on the implantation rate and the likelihood of pregnancy. In addition, being on a weight-loss diet had a negative influence on implantation rate. Our evidence suggests a possible relationship between environmental factors and ovary biology. Copyright © 2015 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Combined effect of radiation and polytriu insecticide on the development of rat embryo

Energy Technology Data Exchange (ETDEWEB)

Abu-Gabal, H A; Eid, F A; Gaber, S H [National Centre for Radiation Research Technology, Faculty of Science for Girls, Al-Azhar University. Cairo (Egypt)

1987-12-31

In the present study 9 groups of pregnant female rats were used. The 1st group served as the control. The 2nd and 3rd groups were orally administered polytrin insecticide:1/100 L D 50 on days 1 and 7 of gestation. Groups 4,5 and 6 were exposed to 2 Gy -rays on day 6, day 12 or days 6 and 12 of gestation. Groups 7,8 and 9 were exposed to 2 Gy gamma -ray on day 6, day 12 or days 6 and 12 during polytrin treatment. The results showed serious abnormal developmental changes as intrauterine death, gross malformations and relation when the embryos or fetuses were maternally exposed to -rays and/or treated with the pyrethroid compound. Moreover the malformations were higher and more drastic in the groups exposed to irradiation during polytrin treatment. These results are very important from the standpoint environmental pollution and radiation protection and safety. figs 23., 8 tabs.

Combined assessment of cyto-, geno-, and embryotoxicity of silica nanoparticles in experiments in vitro

International Nuclear Information System (INIS)

Protasova, G A; Popov, V B; Shabasheva, L V; Strekalovskii, I V; Panferova, Y A; Khlebnikova, N S; Radilov, A A

2017-01-01

A complex of express methods for cyto-, geno-, and embryotoxicity assessment of nanoparticles (NPs), based on the simultaneous use of three models: the cultures of pre- and post-implantation embryos of mice (egg cleavage, blastocyst formation, 1−5 day of development) and rats (head fold−neurulation stages, 30 somite pairs, 9.5−11.5 day of development), as well as human peripheral blood lymphocytes, is developed. Pathogenic (dysmorphogenic) effects of SiO 2 NPs in concentrations of 100 and 200 µg/mL on early embryo development processes were established. The cytogenetic methods used for cyto-, geno-, and mutagenicity assessment (micronuclear test under cytokinesis-block conditions, chromosomal aberrations test, and DNA comet assay) revealed no effects of SiO 2 NPs in concentrations 200, 100, and 20 µg/mL. (paper)

Modifying effects of pre-existing fibrosis in rats exposed to aerosols of {sup 239}PuO{sub 2}. II

Energy Technology Data Exchange (ETDEWEB)

Lundgren, D L; Mauderly, J L; Gillett, N A; Hahn, F F

1988-12-01

We have initiated a study using rats to determine the modifying effects of pre-existing pulmonary fibrosis on the retention and biological effects of inhaled {sup 239}PuO{sub 2}. Pulmonary fibrosis was induced by intratracheal instillation of 8.5 IU/kg body weight of bleomycin at 45 to 49 days before inhalation exposure to an aerosol of {sup 239}PuO{sub 2}. The clearance of {sup 239}Pu from the lungs of rats was decreased significantly (p < 0.01) in rats with pre-existing pulmonary fibrosis compared with controls. Respiratory function, lung morphometric measurements and histological evaluations were all consistent with the presence of mild pulmonary fibrosis in the rats treated with bleomycin. Pre-existing pulmonary fibrosis resulted in an increased retention of the initial lung burdens of {sup 239}Pu, apparently by entrapping the particles in fibrotic areas of the lung. The life span of the rats with pulmonary fibrosis was decreased by up to 25% compared with control rats having similar initial lung burdens of {sup 239}Pu. (author)

Prolonged survival of isolated rat islet allografts pre-irradiated with X-rays

Energy Technology Data Exchange (ETDEWEB)

Kimura, Toshihisa; Note, Masayuki; Nakagawara, Gizo (Fukui Medical School, Matsuoka (Japan)); Kojima, Yasuhiko

1994-04-01

Prior to transplantation of islets, pre-incubation, or pre-irradiation may suppress the immunogenicity of islet cells without suppressing islet function. In the presently described experiments we investigated the use of X-ray irradiation prior to transplantation to reduce islet immunogenicity. To determine whether or not the islet function was reduced after irradiation, ACI rat islets were transplanted into the subrenal capsules of isogeneic rats which had been diabetic and examined the blood glucose level over a period of 40 days. The results indicated that irradiation injury was dose-dependent and that islets irradiated with over 80 Gy lost their function. Next, allogeneic transplantation was performed using the model of ACI rats to Lewis rats without the use of any immunosuppressive agent. Non-irradiated islets were rejected within 7 days. However 20 Gy or 40 Gy irradiated islets prolonged survival (18.7[+-]5.8 days (n=6) and 26.7[+-]10.0 days (n=6), respectively). To determine the basis for this effect, MHC expression of islets was examined by the immunoperoxidase technique. Immunohistologic studies showed that 40 Gy-irradiated islets were depleted of Class II antigen positive cells while Class I antigen expression was unchanged. These results suggest that the prolongation of islets survival by X-ray irradiation may possibly be due to, in part, the depletion of donor Class II antigen positive cells. (author).

Blastomere nucleation: Predictive factors and influence of blastomere with no apparent nuclei on blastocyst development and implantation.

Science.gov (United States)

Setti, Amanda Souza; Figueira, Rita Cássia Sávio; Braga, Daniela Paes de Almeida Ferreira; Iaconelli, Assumpto; Borges, Edson

2018-06-01

To investigate whether embryos presenting blastomere(s) with no apparent nucleus (BNAN) on days 2 and 3 are more likely to fail to develop into blastocysts, hatch and implant. A total of 5705 zygotes obtained from 743 intracytoplasmic sperm injection (ICSI) cycles were analyzed. The presence and incidence of BNAN on days 2 and 3 of embryo development were recorded and then associated with ICSI outcomes. The occurrence of BNAN on day 2 of embryo development was determinant to the decreased odds of blastocyst formation (OR: 0.57, CI: 0.50-0.65), quality (OR: 0.56, CI: 0.43-0.73) and hatching status (OR: 0.66, CI: 0.50-0.87). The presence of BNAN on day 3 of embryo development was determinant to the decreased odds of blastocyst formation (OR: 0.67, CI: 0.58-0.78) and hatching status (OR: 0.61, CI: 0.45-0.83). The occurrence of BNAN on day 2 of embryo development was determinant to the decreased odds of blastocyst implantation (OR: 0.50, CI: 0.27-0.94). The presence of BNAN on day 2 or day 3 reduces development to blastocyst stage, hatching and implantation. Careful nuclear observation, taking into account the absence of blastomere nucleus, should be part of the strategies used for embryo selection.

Influence of Piezosurgery on Bone Healing around Titanium Implants: A Histological Study in Rats.

Science.gov (United States)

Sirolli, Marcelo; Mafra, Carlos Eduardo Secco; Santos, Rodrigo Albuquerque Basílio Dos; Saraiva, Luciana; Holzhausen, Marinella; César, João Batista

2016-01-01

The aim of this study was to evaluate histomorphometrically the influence of two techniques of dental implant site preparation on bone healing around titanium implants. Fifteen male Wistar rats (±300 g) were used in the study. Each tibia was randomly assigned to receive the implant site preparation either with a conventional drilling technique (control - DRILL group) or with a piezoelectric device (PIEZO group). The animals were sacrificed after 30 days and then the following histomorphometric parameters were evaluated (percentage) separately for cortical and cancellous regions: proportion of mineralized tissue (PMT) adjacent to implant threads (500 μm adjacent); bone area within the threads (BA) and bone-implant contact (BIC). The results demonstrated that there were no statistically significant differences between both groups for cancellous BIC (p>0.05) and cortical PMT (p>0.05). On the other hand, a higher percentage of BA was observed in the PIEZO group in the cortical (71.50±6.91 and 78.28±4.38 for DRILL and PIEZO groups, respectively; ppiezosurgery also showed higher PMT values in the cancellous zone (9.35±5.54 and 18.72±13.21 for DRILL and PIEZO groups, respectively; ppiezosurgery was beneficial to bone healing rates in the cancellous bone region, while the drill technique produced better results in the cortical bone.

Embryonic Stem Cell Proteins and MicroRNAs in the Etiology of Germ Cell Cancer

NARCIS (Netherlands)

R. Eini (Ronak)

2013-01-01

textabstractIn the early 1980s, a population of unique cells was isolated from the inner cell mass (ICM) of the mouse pre-implantation embryo named embryonic stem (ES) cells. These cells were generated by removing the ICM from pre-implantation blastocysts. The resulting cells were found to be

Effects of mechanical repetitive load on bone quality around implants in rat maxillae.

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Yusuke Uto

Full Text Available Greater understanding and acceptance of the new concept "bone quality", which was proposed by the National Institutes of Health and is based on bone cells and collagen fibers, are required. The novel protein Semaphorin3A (Sema3A is associated with osteoprotection by regulating bone cells. The aims of this study were to investigate the effects of mechanical loads on Sema3A production and bone quality based on bone cells and collagen fibers around implants in rat maxillae. Grade IV-titanium threaded implants were placed at 4 weeks post-extraction in maxillary first molars. Implants received mechanical loads (10 N, 3 Hz for 1800 cycles, 2 days/week for 5 weeks from 3 weeks post-implant placement to minimize the effects of wound healing processes by implant placement. Bone structures, bone mineral density (BMD, Sema3A production and bone quality based on bone cells and collagen fibers were analyzed using microcomputed tomography, histomorphometry, immunohistomorphometry, polarized light microscopy and birefringence measurement system inside of the first and second thread (designated as thread A and B, respectively, as mechanical stresses are concentrated and differently distributed on the first two threads from the implant neck. Mechanical load significantly increased BMD, but not bone volume around implants. Inside thread B, but not thread A, mechanical load significantly accelerated Sema3A production with increased number of osteoblasts and osteocytes, and enhanced production of both type I and III collagen. Moreover, mechanical load also significantly induced preferential alignment of collagen fibers in the lower flank of thread B. These data demonstrate that mechanical load has different effects on Sema3A production and bone quality based on bone cells and collagen fibers between the inside threads of A and B. Mechanical load-induced Sema3A production may be differentially regulated by the type of bone structure or distinct stress distribution

High risk men's perceptions of pre-implantation genetic diagnosis for hereditary breast and ovarian cancer.

Science.gov (United States)

Quinn, Gwendolyn P; Vadaparampil, Susan T; Miree, Cheryl A; Lee, Ji-Hyun; Zhao, Xiuhua; Friedman, Susan; Yi, Susan; Mayer, James

2010-10-01

Pre-implantation genetic diagnosis (PGD) is an assisted reproductive technology procedure which provides parents with the option of conducting genetic analyses to determine if a mutation is present in an embryo. Though studies have discussed perceptions of PGD from a general population, couples or high-risk women, no studies to date have specifically examined PGD usage among men. This study sought to explore perceptions and attitudes towards PGD among males who either carry a BRCA mutation or have a partner or first degree relative with a BRCA mutation. A cross-sectional survey was conducted among 228 men visiting the Facing Our Risk of Cancer Empowered or Craigslist website. Eligibility criteria included men who self-reported they had been tested for a BRCA mutation or had a partner or first degree relative tested for a BRCA mutation. A 41-item survey assessed socio-demographic, clinical characteristics, PGD knowledge and attitudinal factors and consideration of the use of PGD. Differences in proportions of subgroups were tested using the Monte Carlo exact test for categorical data. A multiple logistic regression model was then built through a backward elimination procedure. Although 80% of men reported being previously unfamiliar with PGD, after learning the definition of PGD, 34% of the 228 respondents then said they would 'ever consider the use of PGD'. Respondents who thought of PGD only in terms of 'health and safety' were almost three times more likely (OR = 2.82; 95% 1.19-6.71) to 'ever consider the use of PGD' compared with respondents who thought of PGD in terms of both 'health and safety', and 'religion and morality'. As with other anonymous web-based surveys, we cannot verify clinical characteristics that may impact consideration of PGD use. Our findings indicate high-risk men need more information about PGD and may benefit from educational materials to assist them in reproductive decision-making.

Efficiency of assisted hatching of the cryopreserved–melted embryos

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V. A. Pitko

2018-04-01

, parameters of positive results of tests on HCG (human chorionic gonadotropin (42 % against 27 % and quantity of clinical pregnancy (35 % against 24 % were statistically higher in the group with assisted hatching comparatively to the control group (P < 0.004; P < 0.001 accordingly. Conclusions. Implantation of the transfered embryos and number of clinical pregnancies were statistically improved due to selection of the optimum freezing conditions and subsequent cultivation and conducting of procedure of mechanical incision of ZP.

Repair of spinal cord injury by implantation of bFGF-incorporated HEMA-MOETACL hydrogel in rats

Science.gov (United States)

Chen, Bo; He, Jianyu; Yang, Hao; Zhang, Qian; Zhang, Lingling; Zhang, Xian; Xie, En; Liu, Cuicui; Zhang, Rui; Wang, Yi; Huang, Linhong; Hao, Dingjun

2015-03-01

There is no effective strategy for the treatment of spinal cord injury (SCI). An appropriate combination of hydrogel materials and neurotrophic factor therapy is currently thought to be a promising approach. In this study, we performed experiments to evaluate the synergic effect of implanting hydroxyl ethyl methacrylate [2-(methacryloyloxy)ethyl] trimethylammonium chloride (HEMA-MOETACL) hydrogel incorporated with basic fibroblast growth factor (bFGF) into the site of surgically induced SCI. Prior to implantation, the combined hydrogel was surrounded by an acellular vascular matrix. Sprague-Dawley rats underwent complete spinal cord transection at the T-9 level, followed by implantation of bFGF/HEMA-MOETACL 5 days after transection surgery. Our results showed that the bFGF/HEMA-MOETACL transplant provided a scaffold for the ingrowth of regenerating tissue eight weeks after implantation. Furthermore, this newly designed implant promoted both nerve tissue regeneration and functional recovery following SCI. These results indicate that HEMA-MOETACL hydrogel is a promising scaffold for intrathecal, localized and sustained delivery of bFGF to the injured spinal cord and provide evidence for the possibility that this approach may have clinical applications in the treatment of SCI.

Implanting Glioblastoma Spheroids into Rat Brains and Monitoring Tumor Growth by MRI Volumetry.

Science.gov (United States)

Löhr, Mario; Linsenmann, Thomas; Jawork, Anna; Kessler, Almuth F; Timmermann, Nils; Homola, György A; Ernestus, Ralf-Ingo; Hagemann, Carsten

2017-01-01

The outcome of patients suffering from glioblastoma multiforme (GBM) remains poor with a median survival of less than 15 months. To establish innovative therapeutical approaches or to analyze the effect of protein overexpression or protein knockdown by RNA interference in vivo, animal models are mandatory. Here, we describe the implantation of C6 glioma spheroids into the rats' brain and how to follow tumor growth by MRI scans. We show that C6 cells grown in Sprague-Dawley rats share several morphologic features of human glioblastoma like pleomorphic cells, areas of necrosis, vascular proliferation, and tumor cell invasion into the surrounding brain tissue. In addition, we describe a method for tumor volumetry utilizing the CISS 3D- or contrast-enhanced T1-weighted 3D sequence and freely available post-processing software.

Chlorhexidine-releasing implant coating on intramedullary nail reduces infection in a rat model

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SM Shiels

2018-03-01

Full Text Available The use of internal intramedullary nails for long bone fracture fixation is a common practice among surgeons. Bacteria naturally attach to these devices, increasing the risk for wound infection, which can result in non- or malunion, additional surgical procedures and extended hospital stays. Intramedullary nail surface properties can be modified to reduce bacterial colonisation and potentially infectious complications. In the current study, a coating combining a non-fouling property with leaching chlorhexidine for orthopaedic implantation was tested. Coating stability and chlorhexidine release were evaluated in vitro. Using a rat model of intramedullary fixation and infection, the effect of the coating on microbial colonisation and fracture healing was evaluated in vivo by quantitative microbiology, micro-computed tomography, plain radiography, three-point bending and/or histology. Low dose systemic cefazolin was administered to increase the similarities to clinical practice, without overshadowing the effect of the anti-infective coating. When introduced into a contaminated wound, the non-fouling chlorhexidine-coated implant reduced the overall bacteria colonisation within the bone and on the implant, reduced the osteolysis and increased the radiographic union, confirming its potential for reducing complications in wounds at high risk of infection. However, when implanted into a sterile wound, non-union increased. Further studies are required to best optimise the anti-microbial effectiveness, while not sacrificing fracture union.

Biocompatibility of a new nanomaterial based on calcium silicate implanted in subcutaneous connective tissue of rats

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Petrović Violeta

2012-01-01

Full Text Available The aim of the study was to investigate rat connective tissue response to a new calcium silicate system 7, 15, 30 and 60 days after implantation. Twenty Wistar albino male rats received two tubes half-filled with a new calcium silicate system (NCSS or MTA in subcutaneous tissue. The empty half of the tubes served as controls. Five animals were sacrificed after 7, 15, 30 and 60 days and samples of the subcutaneous tissue around implanted material were submitted to histological analysis. The intensity of inflammation was evaluated based on the number of inflammatory cells present. Statistical analysis was performed using one way ANOVA and Holm Sidak's multiple comparison tests. Mild to moderate inflammatory reaction was observed after 7, 15 and 30 days around a NCSS while mild inflammatory reaction was detected after 60 days of implantation. In the MTA group, mild to moderate inflammatory reaction was found after 7 and 15 days while mild inflammatory reaction was present after 30 and 60 days. There was no statistically significant difference in the intensity of inflammatory reactions between the tested materials and control groups in any experimental period (ANOVA p>0.05. Regarding the intensity of inflammatory reactions at different experimental periods, a statistically significant difference was observed between 7 and 30 days, 7 and 60 days and 15 to 60 days for both materials. For the controls, a statistically significant difference was found between 7 and 60 days and 15 and 60 days of the experiment (Holm Sidak < p 0.001. Subcutaneous tissue of rats showed good tolerance to a new calcium silicate system. Inflammatory reaction was similar to that caused by MTA. [Projekat Ministarstva nauke Republike Srbije, br. 172026

Critical reappraisal of embryo quality as a predictive parameter for pregnancy outcome: a pilot study.

Science.gov (United States)

Campo, R; Binda, M M; Van Kerkhoven, G; Frederickx, V; Serneels, A; Roziers, P; Lopes, A S; Gordts, S; Puttemans, P; Gordts, S

2010-01-01

Pilot study to analyse the efficacy and embryo morphology using a new human embryo culture medium (GM501) versus the conventional used medium (ISM1). Over a four-month period, all patients at the Leuven Institute of Fertility and Embryology (LIFE) were -randomly allocated to have their embryos cultured in either the standard sequential culture medium ISM1 (control) or in a new universal medium (GM501) (study group). Primary outcome parameters were clinical pregnancy and live birth rate. The secondary outcome parameter was the correlation of embryo fragmentation rate with pregnancy outcome. We did not observe any differences between the ISM1 control group and GM501 study group with regard to fertilization, pregnancy, implantation rates, ongoing pregnancy, and babies born. The number of embryos with a minimal fragmentation rate (less than 30%) was significantly higher in the GM501 study group. Although a significant higher embryo fragmentation rate was seen in In vitro culture of embryos in GM501, pregnancy outcome results were comparable to those of embryos cultured in ISM1. According to our results the value of embryo morphological criteria as a parameter for pregnancy outcome should be examined and discussed again.

Can pre-implantation biopsies predict renal allograft function in pediatric renal transplant recipients?

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Jameela A. Kari

2015-11-01

Full Text Available Objectives: To determine the utility of pre-implantation renal biopsy (PIB to predict renal allograft outcomes. Methods: This is a retrospective review of all patients that underwent PIB from January 2003 to December 2011 at the Great Ormond Street Hospital for Children in London, United Kingdom. Thirty-two male patients (56% aged 1.5-16 years (median: 10.2 at the time of transplantation were included in the study and followed-up for 33 (6-78 months. The results were compared with 33 controls. Results: The PIB showed normal histopathological findings in 13 patients (41%, mild chronic vascular changes in 8 (25%, focal tubular atrophy in one, moderate to severe chronic vascular change in 3, mild to moderate acute tubular damage in 6, and tissue was inadequate in one subject. Delayed graft function (DGF was observed in 3 patients; 2 with vascular changes in PIB, and one with normal histopathological findings. Two subjects with PIB changes lost their grafts. The estimated glomerular filtration rate at 3-, and 6-months post-transplantation was lower in children with abnormal PIB changes compared with those with normal PIB. There was one case of DGF in the control group, and 4 children lost their grafts including the one with DGF. Conclusion: Pre-implantation renal biopsy can provide important baseline information of the graft with implications on subsequent medical treatment for pediatric renal transplant recipients.

Genetic effects on mammalian development during and after implantation

Energy Technology Data Exchange (ETDEWEB)

Pedersen, R A; Spindle, A I

1976-05-01

The effects of gene expression on early mouse embryo differentiation were investigated by measuring the developmental rate of embryos homozygous for the yellow allele (A/sup y/) of the agouti locus and by determining the growth response of normal embryos treated with 5-bromodeoxyuridine (Brd U). The earliest effect of A/sup y/ homozygosity observed in cultured embryos was a delay in cleavage from the 2-cell to the 4-cell stage. Although A/sup y//A/sup y/ embryos were capable of some post-blastocyst development in vitro if the zona pellucida was removed, their inner cell masses disappeared during attachment and trophoblast outgrowth. The period of gene expression that is responsible for the abnormal development of A/sup y//A/sup y/ embryos may thus be between the 2-cell stage and implantation. It is therefore suggested that gene expression necessary for in vitro hatching, attachment, trophoblast outgrowth and inner cell mass development occurs between the morula and late blastocyst stages.

Exercise training prevents the attenuation of anesthetic pre-conditioning-mediated cardioprotection in diet-induced obese rats.

Science.gov (United States)

Li, L; Meng, F; Li, N; Zhang, L; Wang, J; Wang, H; Li, D; Zhang, X; Dong, P; Chen, Y

2015-01-01

Obesity abolishes anesthetic pre-conditioning-induced cardioprotection due to impaired reactive oxygen species (ROS)-mediated adenosine monophosphate-activated protein kinase (AMPK) pathway, a consequence of increased basal myocardial oxidative stress. Exercise training has been shown to attenuate obesity-related oxidative stress. This study tests whether exercise training could normalize ROS-mediated AMPK pathway and prevent the attenuation of anesthetic pre-conditioning-induced cardioprotection in obesity. Male Sprague-Dawley rats were divided into lean rats fed with control diet and obese rats fed with high-fat diet. After 4 weeks of feeding, lean and obese rats were assigned to sedentary conditions or treadmill exercise for 8 weeks. There was no difference in infarct size between lean sedentary and obese sedentary rats after 25 min of myocardial ischemia followed by 120 min reperfusion. In lean rats, sevoflurane equally reduced infarct size in lean sedentary and lean exercise-trained rats. Molecular studies revealed that AMPK activity, endothelial nitric oxide synthase, and superoxide production measured at the end of ischemia in lean rats were increased in response to sevoflurane. In obese rats, sevoflurane increased the above molecular parameters and reduced infarct size in obese exercise-trained rats but not in obese sedentary rats. Additional study showed that obese exercise-trained rats had decreased basal oxidative stress than obese sedentary rats. The results indicate that exercise training can prevent the attenuation of anesthetic cardioprotection in obesity. Preventing the attenuation of this strategy may be associated with reduced basal oxidative stress and normalized ROS-mediated AMPK pathway, but the causal relationship remains to be determined. © 2014 The Acta Anaesthesiologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Histopathological analysis of pre-implantation donor kidney biopsies: association with graft survival and function in one year post-transplantation

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Karla Lais Pêgas

2014-04-01

Full Text Available Introduction: Pre-implantation kidney biopsy is a decision-making tool when considering the use of grafts from deceased donors with expanded criteria, implanting one or two kidneys and comparing this to post-transplantation biopsies. The role of histopathological alterations in kidney compartments as a prognostic factor in graft survival and function has had conflicting results. Objective: This study evaluated the prevalence of chronic alterations in pre-implant biopsies of kidney grafts and the association of findings with graft function and survival in one year post-transplant. Methods: 110 biopsies were analyzed between 2006 and 2009 at Santa Casa de Porto Alegre, including live donors, ideal deceased donors and those with expanded criteria. The score was computed according to criteria suggested by Remuzzi. The glomerular filtration rate (GFR was calculated using the abbreviated MDRD formula. Results: No statistical difference was found in the survival of donors stratified according to Remuzzi criteria. The GFR was significantly associated with the total scores in the groups with mild and moderate alterations, and in the kidney compartments alone, by univariate analysis. The multivariate model found an association with the presence of arteriosclerosis, glomerulosclerosis, acute rejection and delayed graft function. Conclusion: Pre-transplant chronic kidney alterations did not influence the post-transplantation one-year graft survival, but arteriosclerosis and glomerulosclerosis is predictive of a worse GFR. Delayed graft function and acute rejection are independent prognostic factors.

Neural network classification of sweet potato embryos

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Molto, Enrique; Harrell, Roy C.

1993-05-01

Somatic embryogenesis is a process that allows for the in vitro propagation of thousands of plants in sub-liter size vessels and has been successfully applied to many significant species. The heterogeneity of maturity and quality of embryos produced with this technique requires sorting to obtain a uniform product. An automated harvester is being developed at the University of Florida to sort embryos in vitro at different stages of maturation in a suspension culture. The system utilizes machine vision to characterize embryo morphology and a fluidic based separation device to isolate embryos associated with a pre-defined, targeted morphology. Two different backpropagation neural networks (BNN) were used to classify embryos based on information extracted from the vision system. One network utilized geometric features such as embryo area, length, and symmetry as inputs. The alternative network utilized polar coordinates of an embryo's perimeter with respect to its centroid as inputs. The performances of both techniques were compared with each other and with an embryo classification method based on linear discriminant analysis (LDA). Similar results were obtained with all three techniques. Classification efficiency was improved by reducing the dimension of the feature vector trough a forward stepwise analysis by LDA. In order to enhance the purity of the sample selected as harvestable, a reject to classify option was introduced in the model and analyzed. The best classifier performances (76% overall correct classifications, 75% harvestable objects properly classified, homogeneity improvement ratio 1.5) were obtained using 8 features in a BNN.

Host inflammatory response to polypropylene implants: insights from a quantitative immunohistochemical and birefringence analysis in a rat subcutaneous model

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Alessandro Prudente

2016-06-01

Full Text Available ABSTRACT Objectives To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. Materials and Methods Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham. The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment; one unstained (birefringence evaluation and four slides for immunohistochemical processing: IL-1 and TNF-α (pro-inflammatory cytokines, MMP-2 (collagen metabolism and CD-31 (angiogenesis. The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. Results A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002. After 4 days, PP presented higher TNF (p=0.0001 immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06 and IL-1 (p=0.08. After 30 days, a reduction of IL-1 (p=0.010 and TNF (p=0.016 for PP and of IL-1 (p=0.010 for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018. Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively. Conclusions The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization.

Healing of experimental femoral defects in rats after implantation of collagen-calcium phosphate biocomposites

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O. V. Korenkov

2017-06-01

Full Text Available The aim of this study was to investigate the healing process of experimental defects of the femoral shaft diaphysis of rats after implantation of osteoplastic material Collapan into its cavity. In experi-mental animals, a perforated defect with diameter of 2.5 mm was created in the medullary canal of the femoral shaft and filled with osteoplastic material Collapan. In control rats, the defect was left un-filled. The bone fragments were examined on the 15th and 30th day by light microscopy morphometry and scanning electron microscopy. It was found that application of osteoplastic material Collapan in the femoral diaphysis defect optimised reparative osteogenesis, showed high biocompatibility, osteo-conductive properties, resorption ability and good integration with tissue-specific structures of the regenerate

A single pre-operative antibiotic dose is as effective as continued antibiotic prophylaxis in implant-based breast reconstruction: A matched cohort study.

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Townley, William A; Baluch, Narges; Bagher, Shaghayegh; Maass, Saskia W M C; O'Neill, Anne; Zhong, Toni; Hofer, Stefan O P

2015-05-01

Infections following implant-based breast reconstruction can lead to devastating consequences. There is currently no consensus on the need for post-operative antibiotics in preventing immediate infection. This study compared two different methods of infection prevention in this group of patients. A retrospective matched cohort study was performed on consecutive women undergoing implant-based breast reconstruction at University Health Network, Toronto (November 2008-December 2012). All patients received a single pre-operative intravenous antibiotic dose. Group A received minimal interventions and Group B underwent maximal prophylactic measures. Patient (age, smoking, diabetes, co-morbidities), oncologic and procedural variables (timing and laterality) were collected. Univariate and multivariate logistic regression were performed to compare outcomes between the two groups. Two hundred and eight patients underwent 647 implant procedures. After matching the two treatment groups by BMI, 94 patients in each treatment group yielding a total of 605 implant procedures were selected for analysis. The two groups were comparable in terms of patient and disease variables. Post-operative wound infection was similar in Group A (n = 11, 12%) compared with Group B (n = 9, 10%; p = 0.8). Univariate analysis revealed only pre-operative radiotherapy to be associated with the development of infection (0.004). Controlling for the effect of radiotherapy, multivariate analysis demonstrated that there was no statistically significant difference between the two methods for infection prevention. Our findings suggest that a single pre-operative dose of intravenous antibiotics is equally as effective as continued antibiotic prophylaxis in preventing immediate infection in patients undergoing implant-based breast reconstructions. Copyright © 2015 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

Single versus dual renal transplantation from donors with significant arteriosclerosis on pre-implant biopsy.

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Kayler, Liise K; Mohanka, Ravi; Basu, Amit; Shapiro, Ron; Randhawa, Parmjeet S

2009-01-01

Transplantation of kidneys from donor with arteriosclerosis seen on pre-implantation biopsy has not been well studied. We retrospectively evaluated 20 dual kidney transplant (DKT) and 28 single (SKT) kidney transplant recipients with >or=12 months follow-up from donors with moderate arteriosclerosis (>or=25% luminal diameter narrowing). Death censored graft survival was 100% and 79%, respectively (p = 0.0339). DKT recipients had significantly lower mean creatinine levels at one, three, six, and nine months and spent somewhat less time on the waiting list (181 +/- 160 vs. 318 +/- 306 d, p = 0.1429). DKT patients received kidneys from significantly older donors (64 +/- 7 vs. 54 +/- 11 yr; p = 0.0012), proportionately more expanded criteria donors (95% vs. 54%; p = 0.0029), and more donors with hypertension (81% vs. 48%, p = 0.0344) and death related to cerebrovascular accident (100% vs. 71%, p = 0.0143); however, more DKT kidneys underwent machine perfusion (95% vs. 57%, p = 0.0068). Baseline recipient variables were comparable between the two groups including age, race, gender, retransplantation, and HLA mismatch. Pre-implant biopsy was notable for similar frequencies of moderate interstitial fibrosis (10% vs. 14%, respectively) and glomerulosclerosis. Among recipients of deceased-donor kidneys with >25% arteriosclerosis, short-term outcomes after DKT were superior to that of SKT grafts. This approach may help to expand the donor-organ pool while optimizing outcomes.

Pre-expansion before risk reducing mastectomy combined with lipomodelling to enhance results from implant based reconstruction.

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Katerinaki, E; Sircar, T; Fatah, F

2012-02-01

Implant based breast reconstruction after risk reducing mastectomy (RRM) is challenging as implants are inadequately covered in their lower pole. Also complication rates with one stage procedures are significant. We describe a novel method of pre-expansion by inserting subpectoral anatomical expandable implants to prepare the breast mound before RRM is carried out. Lipomodelling is later used to enhance the overall result of the reconstruction. Six patients who were BRCA 1 mutation carriers requested bilateral RRM. Median age was 31. All were non-smokers. Anatomical expandable implants were inserted via an inframammary incision and were expanded in the next two months. RRM was carried out 6-13 months later and was followed by lipomodelling seven months afterwards. Four patients had a nipple sparing mastectomy. In three cases access was via the previous inframammary scar. In all cases complete implant cover had been achieved. There were no early postoperative complications but one patient has developed grade III capsular contracture on the side of previous wide local tumour excision and radiotherapy. Five patients have so far undergone post-mastectomy lipomodelling to improve breast contour. In conclusion, we believe that our technique provides better aesthetic results with reduced complication rates for these patients. Copyright © 2011 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

[Limitations and controversies in determining the predictive value of oocyte and embryo morphology criteria].

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Figueira, Rita de Cássia Savio; Aoki, Tsutomu; Borges Junior, Edson

2015-11-01

In order to increase the success rate of in vitro fertilization cycles, several studies have focused on the identification of the embryo with higher implantation potential. Despite recent advances in the reproductive medicine, based on the OMICs technology, routinely applicable methodologies are still needed. Thus, in most fertilization centers embryo selection for transfer is still based on morphological parameters evaluated under light microscopy. Several morphological parameters may be evaluated, ranging from the pronuclear to blastocyst stage. In general, despite the day of transfer, some criteria are suggested to present a predictive value for embryo viability when analyzed independently or combined. However, the subjectivity of morphological evaluation, as well as the wide diversity of embryo classification systems used by different fertilization centers shows contrasting results, making the implementation of a consensus regarding different morphological criteria and their predictive value a difficult task. The optimization of embryo selection represents a large potential to increase treatment success rates, allowing the transfer of a reduced number of embryos and minimizing the risks of multiple pregnancy.

Warfarin affects acute inflammatory response induced by subcutaneous polyvinyl sponge implantation in rats.

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Mirkov, Ivana; Popov Aleksandrov, Aleksandra; Demenesku, Jelena; Ninkov, Marina; Mileusnic, Dina; Kataranovski, Dragan; Kataranovski, Milena

2017-09-01

Warfarin (WF) is an anticoagulant which also affects physiological processes other than hemostasis. Our previous investigations showed the effect of WF which gained access to the organism via skin on resting peripheral blood granulocytes. Based on these data, the aim of the present study was to examine whether WF could modulate the inflammatory processes as well. To this aim the effect of WF on the inflammatory response induced by subcutaneous sponge implantation in rats was examined. Warfarin-soaked polyvinyl sponges (WF-sponges) were implanted subcutaneously and cell infiltration into sponges, the levels of nitric oxide (NO) and inflammatory cytokines tumor necrosis factor (TNF) and interleukin-6 (IL-6) production by sponge cells were measured as parameters of inflammation. T cell infiltration and cytokine interferon-γ (IFN-γ), interleukin-17 (IL-17) and interleukin-10 (IL-10) were measured at day 7 post implantation. Warfarin exerted both stimulatory and suppressive effects depending on the parameter examined. Flow cytometry of cells recovered from sponges showed higher numbers of granulocytes (HIS48 + cells) at days 1 and 3 post implantation and CD11b + cells at day 1 compared to control sponges. Cells from WF-sponges had an increased NO production (Griess reaction) at days 1 and 7. In contrast, lower levels of TNF (measured by ELISA) production by cells recovered from WF-soaked sponges were found in the early (day one) phase of reaction with unchanged levels at other time points. While IL-6 production by cells recovered from WF-soaked sponges was decreased at day 1, it was increased at day 7. Higher T cell numbers were noted in WF sponges at day 7 post implantation, and recovered cells produced more IFN-γ and IL-17, while IL-10 production remained unchanged. Warfarin affects some of the parameters of inflammatory reaction induced by subcutaneous polyvinyl sponge implantation. Differential (both stimulatory as well as inhibitory) effects of WF on

Effects of single dose GnRH agonist as luteal support on pregnancy outcome in frozen-thawed embryo transfer cycles: an RCT

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Robab Davar

2015-08-01

Full Text Available Background: There is no doubt that luteal phase support is essential to enhance the reproductive outcome in IVF cycles. In addition to progesterone and human chorionic gonadotropin, several studies have described GnRH agonists as luteal phase support to improve implantation rate, pregnancy rate and live birth rate, whereas other studies showed dissimilar conclusions. All of these studies have been done in fresh IVF cycles. Objective: To determine whether an additional GnRH agonist administered at the time of implantation for luteal phase support in frozen-thawed embryo transfer (FET improves the embryo developmental potential. Materials and Methods: This is a prospective controlled trial study in 200 FET cycles, patients were randomized on the day of embryo transfer into group 1 (n=100 to whom a single dose of GnRH agonist (0.1 mg triptorelin was administered three days after transfer and group 2 (n=100, who did not receive agonist. Both groups received daily vaginal progesterone suppositories plus estradiol valerate 6 mg daily. Primary outcome measure was clinical pregnancy rate. Secondary outcome measures were implantation rate, chemical, ongoing pregnancy rate and abortion rate. Results: A total of 200 FET cycles were analyzed. Demographic data and embryo quality were comparable between two groups. No statistically significant difference in clinical and ongoing pregnancy rates was observed between the two groups (26% versus 21%, p=0.40 and 21% versus 17%, p=0.37, respectively. Conclusion: Administration of a subcutaneous GnRH agonist at the time of implantation does not increase clinical or ongoing pregnancy.

Post-implantation mortality of in vitro produced embryos is associated with DNA methyltransferase 1 dysfunction in sheep placenta.

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Ptak, Grazyna Ewa; D'Agostino, Antonella; Toschi, Paola; Fidanza, Antonella; Zacchini, Federica; Czernik, Marta; Monaco, Federica; Loi, Pasqualino

2013-02-01

Is DNA methyltransferase 1 (DNMT1) dysfunction involved in epigenetic deregulation of placentae from embryos obtained by assisted reproduction technologies (ARTs)? DNMT1 expression in growing placentae of in vitro produced (IVP) embryos is compromised and associated with pregnancy loss. DNMT1 maintains the methylation profile of genes during cell division. The methylation status of genes involved in placenta development is altered in embryos obtained in vitro. Disturbances in the epigenetic regulation of gene expression during placentogenesis could be involved in the frequent developmental arrest and loss of IVP embryos. Forty sheep were naturally mated (Group 1, CTR). IVP blastocysts (2-4 per ewe) were surgically transferred to the remaining 46 recipient sheep 6 days after oestrus (Group 2). Twenty-one recipients from Group 1 and 27 recipients from Group 2 were allowed to deliver in order to compare embryo survival in both groups at term (150 days). From the remaining recipients (n = 38), fetuses and placentae of both groups were recovered by paramedian laparotomy at Days 20, 22, 24, 26 and 28 of gestation. Immediately after collection, early placental tissues (chorion-allantois) were snap frozen in liquid nitrogen and DNMT1 expression and activity was evaluated. mRNA levels (for DNMT1, HDAC2, PCNA, DMAP1, MEST, IGF2, CDKN1C, H19) and the methylation status of H19 were also analyzed. Furthermore, embryo size and survival rate were measured. Our study shows that DNMT1 expression was reduced in early placentae from sheep IVP embryos. This reduction was associated with growth arrest and subsequent death of the sheep embryos. Conversely, normal levels of DNMT1 and its cofactors were observed in placentae from IVP embryos that survived this developmental bottleneck. Although DNA methylation machinery was severely compromised in IVP placentae only up to Day 24, the low DNMT1 enzymatic activity that persisted after this stage in IVP placentae was not lethal for the

Effect of variable scanning protocolson the pre-implant site evaluation of the mandible in reformatted computed tomography

International Nuclear Information System (INIS)

Kim, Kee Deog; Park, Chang Seo

1999-01-01

To evaluate the effect of variable scanning protocols of computed tomography for evaluation of pre-implant site of the mandible through the comparison of the reformatted cross-sectional images of helical CT scans obtained with various imaging parameters versus those of conventional CT scans. A dry mandible was imaged using conventional nonoverlapped CT scans with 1 mm slice thickness and helical CT scans with 1 mm slice thickness and pitches of 1.0, 1.5, 2.0, 2.5 and 3.0. All helical images were reconstructed at reconstruction interval of 1 mm. DentaScan reformatted images were obtained to allow standardized visualization of cross-sectional images of the mandible. The reformatted images were reviewed and measured separately by 4 dental radiologists. The image qualities of continuity of cortical outline, trabecular bone structure and visibility of the mandibular canal were evaluated and the distance between anatomic structures were measured by 4 dental radiologists. On image qualities of continuity of cortical outline, trabecular bone structure and visibility of the mandibular canal and in horizontal measurement, there was no statistically significant difference among conventional and helical scans with pitches of 1.0, 1.5 and 2.0. In vertical measurement, there was no statistically significant difference among the conventional and all imaging parameters of helical CT scans with pitches of 1.0, 1.5, 2.0, 2.5 and 3.0. The images of helical CT scans with 1 mm slice thickness and pitches of 1.0, 1.5 and 2.0 are as good as those of conventional CT scans with 1 mm slice thickness for evaluation of pre-dental implant site of the mandible. Considering the radiation dose and patient comfort, helical CT scans with 1 mm slice thickness and pitch of 2.0 is recommended for evaluation of pre-implant site of the mandible.

Comparing Binaural Pre-processing Strategies II: Speech Intelligibility of Bilateral Cochlear Implant Users.

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Baumgärtel, Regina M; Hu, Hongmei; Krawczyk-Becker, Martin; Marquardt, Daniel; Herzke, Tobias; Coleman, Graham; Adiloğlu, Kamil; Bomke, Katrin; Plotz, Karsten; Gerkmann, Timo; Doclo, Simon; Kollmeier, Birger; Hohmann, Volker; Dietz, Mathias

2015-12-30

Several binaural audio signal enhancement algorithms were evaluated with respect to their potential to improve speech intelligibility in noise for users of bilateral cochlear implants (CIs). 50% speech reception thresholds (SRT50) were assessed using an adaptive procedure in three distinct, realistic noise scenarios. All scenarios were highly nonstationary, complex, and included a significant amount of reverberation. Other aspects, such as the perfectly frontal target position, were idealized laboratory settings, allowing the algorithms to perform better than in corresponding real-world conditions. Eight bilaterally implanted CI users, wearing devices from three manufacturers, participated in the study. In all noise conditions, a substantial improvement in SRT50 compared to the unprocessed signal was observed for most of the algorithms tested, with the largest improvements generally provided by binaural minimum variance distortionless response (MVDR) beamforming algorithms. The largest overall improvement in speech intelligibility was achieved by an adaptive binaural MVDR in a spatially separated, single competing talker noise scenario. A no-pre-processing condition and adaptive differential microphones without a binaural link served as the two baseline conditions. SRT50 improvements provided by the binaural MVDR beamformers surpassed the performance of the adaptive differential microphones in most cases. Speech intelligibility improvements predicted by instrumental measures were shown to account for some but not all aspects of the perceptually obtained SRT50 improvements measured in bilaterally implanted CI users. © The Author(s) 2015.

The interaction between Xe and F in Si (1 0 0) pre-amorphised with 20 keV Xe and implanted with low energy BF{sub 2}

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Werner, M. [Joule Physics Laboratory, Institute of Materials Research, University of Salford, Salford M54WT (United Kingdom)]. E-mail: m.werner@pgr.salford.ac.uk; Berg, J.A. van den [Joule Physics Laboratory, Institute of Materials Research, University of Salford, Salford M54WT (United Kingdom); Armour, D.G. [Joule Physics Laboratory, Institute of Materials Research, University of Salford, Salford M54WT (United Kingdom); Carter, G. [Joule Physics Laboratory, Institute of Materials Research, University of Salford, Salford M54WT (United Kingdom); Feudel, T. [AMD Saxony LLC and Co. KG, Wilschdorfer Landstrasse, 101 D-01109 Dresden (Italy); Herden, M. [AMD Saxony LLC and Co. KG, Wilschdorfer Landstrasse, 101 D-01109 Dresden (Italy); Bersani, M. [ITC IRST, 38050 Povo, Trento (Italy); Giubertoni, D. [ITC IRST, 38050 Povo, Trento (Italy); Bailey, P. [CCLRC Daresbury Laboratory, Daresbury WA44A (United Kingdom); Noakes, T.C.Q. [CCLRC Daresbury Laboratory, Daresbury WA44A (United Kingdom)

2004-12-15

The pre-amorphisation of Si by Xe{sup +} ions, before source/drain and extension implants, is an attractive alternative to Ge{sup +} or Si{sup +}, as it produces sharper amorphous/crystalline interfaces. Si (1 0 0) samples pre-amorphised with 20 keV Xe{sup +} to a nominal dose of 2E14 cm{sup -2} were implanted with 1 and 3 keV BF{sub 2} {sup +} to doses of 7E14 cm{sup -2}. Samples were annealed at temperatures ranging from 600 to 1130 deg. C and investigated by medium energy ion scattering (MEIS) and secondary ion mass spectrometry (SIMS). Following annealing, it was observed that implanted Xe has interacted with F originating from the BF{sub 2} {sup +} implant. MEIS studies showed that for all annealing conditions, approximately half of the Xe accumulated at depths of 7 nm for the 1 keV and at 13 nm for the 3 keV BF{sub 2} {sup +} implant. This equates to the end of range of B and F within the amorphous Si. SIMS showed that in the pre-amorphised samples, approximately 10% of the F migrates into the bulk and is trapped at the same depth in a {approx}1:1 ratio to Xe. A small fraction of the implanted B is also trapped. The effect is interpreted in terms of the formation of a defect structure within the amorphised Si, leading to F stabilised Xe bubble or XeF compound formation.

Podoplanin immunopositive lymphatic vessels at the implant interface in a rat model of osteoporotic fractures.

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Katrin Susanne Lips

Full Text Available Insertion of bone substitution materials accelerates healing of osteoporotic fractures. Biodegradable materials are preferred for application in osteoporotic patients to avoid a second surgery for implant replacement. Degraded implant fragments are often absorbed by macrophages that are removed from the fracture side via passage through veins or lymphatic vessels. We investigated if lymphatic vessels occur in osteoporotic bone defects and whether they are regulated by the use of different materials. To address this issue osteoporosis was induced in rats using the classical method of bilateral ovariectomy and additional calcium and vitamin deficient diet. In addition, wedge-shaped defects of 3, 4, or 5 mm were generated in the distal metaphyseal area of femur via osteotomy. The 4 mm defects were subsequently used for implantation studies where bone substitution materials of calcium phosphate cement, composites of collagen and silica, and iron foams with interconnecting pores were inserted. Different materials were partly additionally functionalized by strontium or bisphosphonate whose positive effects in osteoporosis treatment are well known. The lymphatic vessels were identified by immunohistochemistry using an antibody against podoplanin. Podoplanin immunopositive lymphatic vessels were detected in the granulation tissue filling the fracture gap, surrounding the implant and growing into the iron foam through its interconnected pores. Significant more lymphatic capillaries were counted at the implant interface of composite, strontium and bisphosphonate functionalized iron foam. A significant increase was also observed in the number of lymphatics situated in the pores of strontium coated iron foam. In conclusion, our results indicate the occurrence of lymphatic vessels in osteoporotic bone. Our results show that lymphatic vessels are localized at the implant interface and in the fracture gap where they might be involved in the removal of

Podoplanin Immunopositive Lymphatic Vessels at the Implant Interface in a Rat Model of Osteoporotic Fractures

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Lips, Katrin Susanne; Kauschke, Vivien; Hartmann, Sonja; Thormann, Ulrich; Ray, Seemun; Kampschulte, Marian; Langheinrich, Alexander; Schumacher, Matthias; Gelinsky, Michael; Heinemann, Sascha; Hanke, Thomas; Kautz, Armin R.; Schnabelrauch, Matthias; Schnettler, Reinhard; Heiss, Christian; Alt, Volker; Kilian, Olaf

2013-01-01

Insertion of bone substitution materials accelerates healing of osteoporotic fractures. Biodegradable materials are preferred for application in osteoporotic patients to avoid a second surgery for implant replacement. Degraded implant fragments are often absorbed by macrophages that are removed from the fracture side via passage through veins or lymphatic vessels. We investigated if lymphatic vessels occur in osteoporotic bone defects and whether they are regulated by the use of different materials. To address this issue osteoporosis was induced in rats using the classical method of bilateral ovariectomy and additional calcium and vitamin deficient diet. In addition, wedge-shaped defects of 3, 4, or 5 mm were generated in the distal metaphyseal area of femur via osteotomy. The 4 mm defects were subsequently used for implantation studies where bone substitution materials of calcium phosphate cement, composites of collagen and silica, and iron foams with interconnecting pores were inserted. Different materials were partly additionally functionalized by strontium or bisphosphonate whose positive effects in osteoporosis treatment are well known. The lymphatic vessels were identified by immunohistochemistry using an antibody against podoplanin. Podoplanin immunopositive lymphatic vessels were detected in the granulation tissue filling the fracture gap, surrounding the implant and growing into the iron foam through its interconnected pores. Significant more lymphatic capillaries were counted at the implant interface of composite, strontium and bisphosphonate functionalized iron foam. A significant increase was also observed in the number of lymphatics situated in the pores of strontium coated iron foam. In conclusion, our results indicate the occurrence of lymphatic vessels in osteoporotic bone. Our results show that lymphatic vessels are localized at the implant interface and in the fracture gap where they might be involved in the removal of lymphocytes, macrophages

Human oocyte oolemma characteristic is positively related to embryo developmental competence after ICSI procedure

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Mohamed A. Danfour

2010-10-01

Conclusion: The current study provides evidence that preselection at a very early stage based on oolemma behavior may be helpful to identify a subgroup of preimplantation embryos with good prognostic to form blastocyst and consequently to implant and to give pregnancy.

Ability of mini-implant-facilitated micro-osteoperforations to accelerate tooth movement in rats.

Science.gov (United States)

Cheung, Tracy; Park, Juyoung; Lee, Deborah; Kim, Catherine; Olson, Jeffrey; Javadi, Shadi; Lawson, Gregory; McCabe, James; Moon, Won; Ting, Kang; Hong, Christine

2016-12-01

Although current techniques for accelerated tooth movement often involve invasive surgical procedures, micro-osteoperforations (MOPs) using mini-implants may facilitate orthodontic tooth movement without raising flaps, reduce surgical risks, and increase patient acceptance. In this study, we evaluated the effectiveness of mini-implant-facilitated MOPs in inducing accelerated tooth movement and investigated the potential risks for root resorption. Five MOPs were placed on the left side around the maxillary first molars in 6 rats using an automated mini-implant driver, whereas the right side received no MOPs as the control. Closed-coiled springs were secured from incisors to first molars for orthodontic tooth movement. Tooth movement was measured, and samples underwent radiologic and histologic analyses. The MOP side exhibited a 1.86-fold increase in the rate of tooth movement with decreased bone density and bone volume around the first molars compared with the control side. Hematoxylin and eosin and tartrate-resistant acid phosphatase analyses showed increased numbers of osteoclasts as well as new bone formation. Three-dimensional volumetric analysis of all 5 roots of the maxillary first molars demonstrated no statistically significant difference in root volumes. Mini-implant-facilitated MOPs accelerated tooth movement without increased risk for root resorption and therefore may become a readily available and efficient treatment option to shorten orthodontic treatment time with improved patient acceptance. Copyright © 2016 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.

The Well-of-the-Well system: an efficient approach to improve embryo development.

Science.gov (United States)

Vajta, Gábor; Korösi, Tamás; Du, Yutao; Nakata, Kumiko; Ieda, Shoko; Kuwayama, Masashige; Nagy, Zsolt Peter

2008-07-01

Transfer of human embryos at the blastocyst stage may offer considerable benefits including an increased implantation rate and a decreased risk of multiple pregnancies; however, blastocyst culture requires an efficient and reliable in-vitro embryo culture system. In this study, the effect of the Well-of-the-Well (WOW) system consisting of microwells formed on the bottom of the culture dish was tested in three mammalian species, including humans. The WOW system resulted in significant improvement when comparing the drops for culture of in-vitro-matured and parthenogenetically activated porcine oocytes, and in-vivo-derived mouse zygotes. In human embryos, using a sibling oocyte design, embryos cultured in WOW developed to the blastocyst stage in a significantly higher proportion than did embryos cultured traditionally (55% in WOW and 37% in conventional culture; P WOW system or in microdrops. Transferable quality blastocyst development (48.9% of cultured zygotes) was observed in the WOW system. Ninety-four blastocysts transferred to 45 patients resulted in clinical pregnancy rates of 48.9%, including nine twin pregnancies, seven single pregnancies, five miscarriages and one ectopic pregnancy. The results indicate that the WOW system provides a promising alternative for microdrop culture of mammalian embryos, including human embryos.

Influence of radiation (Co{sub 60}) in pre-implant rabbit embryos: effect on mitotic index and embryonic pole malformations; Influencia da radiacao ionizante (bomba de cobalto) em embrioes de coelha na fase de pre-implantacao: influencia no indice mitotico e malformacoes no polo embrionario

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Approbato, M.S.; Moura, K.K.V.O.; Florencio, R.S.; Cunha Junior, C.; Garcia, R.; Faria, R.S.; Benedetti, L.N.; Goulart, F.B. [Goias Univ., Goiania, GO (Brazil). Faculdade de Medicina. Dept. de Ginecologia e Obstetricia

1995-03-01

We studied the effect of ionizing irradiation on 12 New Zealand rabbits (65 embryos), at three different times: at match time (zero hour), two days after and four days after, with two different irradiation doses: five c Gy and ten c Gy. Six rabbits (36 blastocysts) were used as controls. the matching instant was the zero hour. Exactly six days after ({+-} 60 minutes) the embryos of each rabbit was picked up by flushing the uterus with culture media. the embryos were fixed in methanol for 48 hours, and colored with acid Mayer hematoxylin. The following embryo parameters were studied: mitotic index; embryonic pole malformations. There were no gross abnormalities of embryo pole. The mitotic index were altered both by the time and doses. (author). 12 refs., 3 figs.

Clinical outcome of fresh and vitrified-warmed blastocyst and cleavage-stage embryo transfers in ethnic Chinese ART patients

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Tong Guo

2012-10-01

Full Text Available Abstract Objectives This study sought to evaluate the outcome of fresh and vitrified-warmed cleavage-stage and blastocyst-stage embryo transfers in patients undergoing ART treatment within an ethnic Chinese population. Study design We compared the clinical results of embryo transfer on the 3rd (cleavage stage or 5th (blastocyst stage day after oocyte retrieval, including clinical pregnancy rates, implantation rates and multiple pregnancy rates. Results Our data showed that blastocyst transfer on day 5 did not significantly increase clinical pregnancy rate (41.07% vs 47.08%, p>0.05 and implantation rate (31.8% vs 31.2%, p>0.05 in patients under 35 years of age, in comparison with day 3 cleavage stage embryo transfer. In patients older than 35 years of age, the clinical pregnancy rate after blastocyst transfer was slightly decreased compared with cleavage stage embryo transfer (33.33% vs 42.31%, p>0.05. Unexpectedly, It was found that vitrified-warmed blastocyst transfer resulted in significantly higher clinical pregnancy rate (56.8% and implantation rate (47% compared with fresh blastocyst transfer in controlled stimulation cycles (41.07% and 31.8%, respectively. For patients under 35 years of age, the cumulative clinical pregnancy rate combining fresh and vitrified-warmed blastocyst transfer cycles were significantly higher compared to just cleavage-stage embryo transfer (70.1% versus 51.8%, p Conclusions In an ethnic Chinese patient population, fresh blastocyst transfer does not significantly increase clinical pregnancy rate. However, subsequent vitrified-warmed blastocyst transfer in a non-controlled ovarian hyperstimulation cycle dramatically improves clinical outcomes. Therefore, blastocyst culture in tandem with vitrified-warmed blastocyst transfer is recommended as a favourable and promising protocol in human ART treatment, particularly for ethnic Chinese patients.

Clinical outcome of fresh and vitrified-warmed blastocyst and cleavage-stage embryo transfers in ethnic Chinese ART patients.

Science.gov (United States)

Tong, Guo Qing; Cao, Shan Ren; Wu, Xun; Zhang, Jun Qiang; Cui, Ji; Heng, Boon Chin; Ling, Xiu Feng

2012-10-05

This study sought to evaluate the outcome of fresh and vitrified-warmed cleavage-stage and blastocyst-stage embryo transfers in patients undergoing ART treatment within an ethnic Chinese population. We compared the clinical results of embryo transfer on the 3rd (cleavage stage) or 5th (blastocyst stage) day after oocyte retrieval, including clinical pregnancy rates, implantation rates and multiple pregnancy rates. Our data showed that blastocyst transfer on day 5 did not significantly increase clinical pregnancy rate (41.07% vs 47.08%, p>0.05) and implantation rate (31.8% vs 31.2%, p>0.05) in patients under 35 years of age, in comparison with day 3 cleavage stage embryo transfer. In patients older than 35 years of age, the clinical pregnancy rate after blastocyst transfer was slightly decreased compared with cleavage stage embryo transfer (33.33% vs 42.31%, p>0.05). Unexpectedly, It was found that vitrified-warmed blastocyst transfer resulted in significantly higher clinical pregnancy rate (56.8%) and implantation rate (47%) compared with fresh blastocyst transfer in controlled stimulation cycles (41.07% and 31.8%, respectively). For patients under 35 years of age, the cumulative clinical pregnancy rate combining fresh and vitrified-warmed blastocyst transfer cycles were significantly higher compared to just cleavage-stage embryo transfer (70.1% versus 51.8%, p<0.05). However, the cumulative multiple pregnancy rates showed no significant difference between the two groups. In an ethnic Chinese patient population, fresh blastocyst transfer does not significantly increase clinical pregnancy rate. However, subsequent vitrified-warmed blastocyst transfer in a non-controlled ovarian hyperstimulation cycle dramatically improves clinical outcomes. Therefore, blastocyst culture in tandem with vitrified-warmed blastocyst transfer is recommended as a favourable and promising protocol in human ART treatment, particularly for ethnic Chinese patients.

Endometrial scratching in women with implantation failure after a first IVF/ICSI cycle; does it lead to a higher live birth rate? The SCRaTCH study: a randomized controlled trial (NTR 5342)

NARCIS (Netherlands)

Hoogenhuijze, N.E. van; Torrance, H.L.; Mol, F.; Laven, J.S.; Scheenjes, E.; Traas, M.A.F.; Janssen, C.; Cohlen, B.; Teklenburg, G.; Bruin, J.P. de; Oppenraaij, R. van; Maas, J.W.; Moll, E.; Fleischer, K.; Hooff, M.H. van; Koning, C.; Cantineau, A.; Lambalk, C.B.; Verberg, M.; Nijs, M.; Manger, A.P.; Rumste, M. van; Voet, L.F. van der; Preys-Bosman, A.; Visser, J.; Brinkhuis, E.; Hartog, J.E. Den; Sluijmer, A.; Jansen, F.W.; Hermes, W.; Bandell, M.L.; Pelinck, M.J.; Disseldorp, J. van; Wely, M. van; Smeenk, J.; Pieterse, Q.D.; Boxmeer, J.C.; Groenewoud, E.R.; Eijkemans, M.J.; Kasius, J.C.; Broekmans, F.J.

2017-01-01

BACKGROUND: Success rates of assisted reproductive techniques (ART) are approximately 30%, with the most important limiting factor being embryo implantation. Mechanical endometrial injury, also called 'scratching', has been proposed to positively affect the chance of implantation after embryo

Endometrial scratching in women with implantation failure after a first IVF/ICSI cycle; does it lead to a higher live birth rate? The SCRaTCH study : A randomized controlled trial (NTR 5342)

NARCIS (Netherlands)

van Hoogenhuijze, N. E.; Torrance, H. L.; Mol, F.; Laven, Joop S E; Scheenjes, E.; Traas, M.A.F.; Janssen, Bert J C; Cohlen, B.; Teklenburg, G.; Bruin, Jacob P; van Oppenraaij, R.; Maas, J. W M; Moll, Etelka; Fleischer, K.; van Hooff, Marcel H; de Koning, C.; Cantineau, A.; Lambalk, Cornelis B.; Verberg, M.F.G.; Nijs, M. M.; Manger, A. P.; Van Rumste, M. M.; Van der Voet, L F; Preys-Bosman, A.; Visser, J.; Brinkhuis, E.A.; den Hartog, J. E.; Sluijmer, A.; Jansen, F. W.; Hermes, W.; Bandell, M. L.; Pelinck, M. J.; van Disseldorp, J.; Wely, M.; Smeenk, J.; Pieterse, Q.D.; Boxmeer, J.C.; Groenewoud, Eva R.; Eijkemans, M. J.C.; Kasius, J. C.; Broekmans, F. J.M.

2017-01-01

Background: Success rates of assisted reproductive techniques (ART) are approximately 30%, with the most important limiting factor being embryo implantation. Mechanical endometrial injury, also called 'scratching', has been proposed to positively affect the chance of implantation after embryo

Treatment of diabetic rats with encapsulated islets.

Science.gov (United States)

Sweet, Ian R; Yanay, Ofer; Waldron, Lanaya; Gilbert, Merle; Fuller, Jessica M; Tupling, Terry; Lernmark, Ake; Osborne, William R A

2008-12-01

Immunoprotection of islets using bioisolator systems permits introduction of allogeneic cells to diabetic patients without the need for immunosuppression. Using TheraCyte immunoisolation devices, we investigated two rat models of type 1 diabetes mellitus (T1DM), BB rats and rats made diabetic by streptozotocin (STZ) treatment. We chose to implant islets after the onset of diabetes to mimic the probable treatment of children with T1DM as they are usually diagnosed after disease onset. We encapsulated 1000 rat islets and implanted them subcutaneously (SQ) into diabetic biobreeding (BB) rats and STZ-induced diabetic rats, defined as two or more consecutive days of blood glucose>350 mg/dl. Rats were monitored for weight and blood glucose. Untreated BB rats rapidly lost weight and were euthanized at >20% weight loss that occurred between 4 and 10 days from implantation. For period of 30-40 days following islet implantation weights of treated rats remained steady or increased. Rapid weight loss occurred after surgical removal of devices that contained insulin positive islets. STZ-treated rats that received encapsulated islets showed steady weight gain for up to 130 days, whereas untreated control rats showed steady weight loss that achieved >20% at around 55 days. Although islet implants did not normalize blood glucose, treated rats were apparently healthy and groomed normally. Autologous or allogeneic islets were equally effective in providing treatment. TheraCyte devices can sustain islets, protect allogeneic cells from immune attack and provide treatment for diabetic-mediated weight loss in both BB rats and STZ-induced diabetic rats.

Distinct differences in global gene expression profiles in non-implanted blastocysts and blastocysts resulting in live birth

DEFF Research Database (Denmark)

Kirkegaard, Kirstine Kjær; Fredsted, Palle Villesen; Jensen, Jacob Malte

2015-01-01

Results from animal models points towards the existence of a gene expression profile that is distinguishably different in viable embryos compared with non-viable embryos. Knowledge of human embryo transcripts is however limited, in particular with regard to how gene expression is related...... to clinical outcome. The purpose of the present study was therefore to determine the global gene expression profiles of human blastocysts. Next Generation Sequencing was used to identify genes that were differentially expressed in non-implanted embryos and embryos resulting in live birth. Three trophectoderm...

Effect of pomegranate juice pre-treatment on the transport of carbamazepine across rat intestine

Directory of Open Access Journals (Sweden)

D Adukondalu

2010-12-01

Full Text Available "n  "nBackground and the purpose of the study: Many drug substances along with a variety of naturally occurring dietary or herbal components interact with the CYP enzyme system.The present study was aimed to investigate the effect of pomegranate juice pre-treatment on the transport of carbamazepine across the rat intestine "nMethods: The transport of carbamazepine across different parts of rat intestine was studied by everted and non-everted sac methods. The control and pomegranate juice (10 ml Kg-1 for 7 days pre-treated rats were sacrificed and isolated the intestine. The sacs of intestine were prepared, treated with carbamazepine solution and then placed in dulbeccos buffer. Samples were collected periodically and the drug content was estimated using HPLC. Results and conclusion: The results show that there was a significant (p<0.05 difference in the transport of carbamazepine from the intestinal sacs of pretreated with pomegranate juice and control. It seems that pomegranatejuice might have induced CYP3A4enzymes and hence drug is extensively metabolized.

Chronic implantation of cuff electrodes on the pelvic nerve in rats is well tolerated and does not compromise afferent or efferent fibre functionality

Science.gov (United States)

Crook, J. J.; Brouillard, C. B. J.; Irazoqui, P. P.; Lovick, T. A.

2018-04-01

Objective. Neuromodulation of autonomic nerve activity to regulate physiological processes is an emerging field. Vagal stimulation has received most attention whereas the potential of modulate visceral function by targeting autonomic nerves within the abdominal cavity remains under-exploited. Surgery to locate intra-abdominal targets is inherently more stressful than for peripheral nerves. Electrode leads risk becoming entrapped by intestines and loss of functionality in the nerve-target organ connection could result from electrode migration or twisting. Since nociceptor afferents are intermingled with similar-sized visceral autonomic fibres, stimulation may induce pain. In anaesthetised rats high frequency stimulation of the pelvic nerve can suppress urinary voiding but it is not known how conscious animals would react to this procedure. Our objective therefore was to determine how rats tolerated chronic implantation of cuff electrodes on the pelvic nerve, whether nerve stimulation would be aversive and whether nerve-bladder functionality would be compromised. Approach. We carried out a preliminary de-risking study to investigate how conscious rats tolerated chronic implantation of electrodes on the pelvic nerve, their responsiveness to intermittent high frequency stimulation and whether functionality of the nerve-bladder connection became compromised. Main results. Implantation of cuff electrodes was well-tolerated. The normal diurnal pattern of urinary voiding was not disrupted. Pelvic nerve stimulation (up to 4 mA, 3 kHz) for 30 min periods evoked mild alerting at stimulus onset but no signs of pain. Stimulation evoked a modest (nerve temperature but the functional integrity of the nerve-bladder connection, reflected by contraction of the detrusor muscle in response to 10 Hz nerve stimulation, was not compromised. Significance. Chronic implantation of cuff electrodes on the pelvic nerve was found to be a well-tolerated procedure in rats and high frequency

Screening of patients for cochlear implant through a questionnaire online. GroupProfile of patients pre-and peri lingual not summoned

Directory of Open Access Journals (Sweden)

Leal, Aquiles Figueiredo

2010-06-01

Full Text Available Introduction: Facilitating access to specialized centers and properly screen patients seeking cochlear implants are critical steps for proper rehabilitation. Objective: To describe the group of patients pre-and peri-lingual is not called for in a service evaluation of cochlear implants. Method: A retrospective study analyzed 401 questionnaires of patients pre-and peri-lingual Web site registered in the Central Brazilian cochlear implant. For the failure to call these patients were used as criteria applied some variables: age, use of hearing aids, speech therapy, duration of deafness, type of progression of hearing loss and type of communication used by the patient. Results: The group of patients with pre-and peri-lingual deafness accounted for 34% of total questionnaires completed during the period. The distribution by age found that 54% of patients were over 17 years, 30% between 9 and 17 years, and remaining less than 9 years. The duration of deafness was higher than 20 years in 50% of patients, between 10 and 20 years by 32% between 5 and 10 years in 9% and between 0 and 5 years in 9%. Regarding the performance of voice rehabilitation 58% of patients had performed and 42% did not. Regarding the mode of communication 49% had global communication, 18% LIBRAS, 6% oral communication, 26% no communication. Conclusion: Advanced age, duration of deafness high, so mostly no oral communication and lack of voice rehabilitation were crucial to the failure to call these patients.

Detrimental effects of rat mesenchymal stromal cell pre-treatment in a model of acute kidney rejection

Directory of Open Access Journals (Sweden)

Martina eSeifert

2012-07-01

Full Text Available Mesenchymal stromal cells (MSC have shown immunomodulatory and tissue repair potential including partial tolerance induction by pre-treatment of donor-specific cells in a rat heart transplantation model. Very recently, we could show that autologous MSC attenuated ischemia reperfusion injury in a highly mismatched donor-recipient rat kidney transplant model. Therefore, we investigated donor-specific MSC pre-treatment in this rat kidney transplantation model to study whether graft function could be improved, or if tolerance could be induced.Donor- and recipient-type MSC or PBS as a control were injected i.v. four days before kidney transplantation. Mycophenolate mofetil (MMF immunosuppression (20 mg/kg body weight was applied for 7 days. Kidney grafts and spleens were harvested between days 8-10 and analyzed by quantitative RT-PCR and immunohistology. In addition, creatinine levels in the blood were measured and serum was screened for the presence of donor-specific antibodies.Surprisingly, application of both donor- and recipient-specific MSC resulted in enhanced humoral immune responses verified by intragraft B cell infiltration and complement factor C4d deposits. Moreover, signs of inflammation and rejection were generally enhanced in both MSC-treated groups relative to PBS control group. Additionally, pre-treatment with donor-specific MSC significantly enhanced the level of donor-specific antibody formation when compared with PBS- or recipient-MSC-treated groups. Pre-treatment with both MSC types resulted in a higher degree of kidney cortex tissue damage and elevated creatinine levels at the time point of rejection. Thus, MSC pre-sensitization in this model impairs the allograft outcome.Our data from this pre-clinical kidney transplantation model indicate that pre-operative MSC administration may not be optimal in kidney transplantation and caution must be exerted before moving forward with clinical studies in order to avoid adverse effects.

Sub-meninges implantation reduces immune response to neural implants.

Science.gov (United States)

Markwardt, Neil T; Stokol, Jodi; Rennaker, Robert L

2013-04-15

Glial scar formation around neural interfaces inhibits their ability to acquire usable signals from the surrounding neurons. To improve neural recording performance, the inflammatory response and glial scarring must be minimized. Previous work has indicated that meningeally derived cells participate in the immune response, and it is possible that the meninges may grow down around the shank of a neural implant, contributing to the formation of the glial scar. This study examines whether the glial scar can be reduced by placing a neural probe completely below the meninges. Rats were implanted with sets of loose microwire implants placed either completely below the meninges or implanted conventionally with the upper end penetrating the meninges, but not attached to the skull. Histological analysis was performed 4 weeks following surgical implantation to evaluate the glial scar. Our results found that sub-meninges implants showed an average reduction in reactive astrocyte activity of 63% compared to trans-meninges implants. Microglial activity was also reduced for sub-meninges implants. These results suggest that techniques that isolate implants from the meninges offer the potential to reduce the encapsulation response which should improve chronic recording quality and stability. Published by Elsevier B.V.