Search results for: single embryo transfer

Authors: Van Huong Do, Simon Walton, German Amaya, Madeline Batsiokis, Sally Catt, Andrew Taylor-Robinson

Abstract:

Identification of the most suitable stages of bovine in vitro-derived blastocysts (early, expanded and hatching) prior to vitrification is a straightforward process that facilitates the decision as to which blastocyst stage to use for transfer of fresh and vitrified embryos. Research on in vitro evaluation of suitable stages has shown that the more advanced developmental stage of blastocysts is recommended for fresh embryo transfer while the earlier stage is proposed for embryo transfer following vitrification. There is, however, limited information on blastocyst stages using in vivo assessment. Hence, the aim of the present study was to determine the optimal stage of a blastocyst for vitrification and embryo transfer through a two-step procedure of embryo transfer followed by pregnancy testing at 35, 60 and 90 days of pregnancy. 410 good quality oocytes aspirated by the ovum pick-up technique from 8 donor cows were subjected to in vitro embryo production, vitrification and embryo transfer. Good quality embryos were selected, subjected to vitrification and embryo transfer. Subsequently, 77 vitrified embryos at different blastocyst stages were transferred to synchronised recipient cows. The overall cleavage and blastocyst rates of oocytes were 68.8% and 41.7%, respectively. In addition, the fertility and blastocyst production of 6 bulls used for in vitro fertilization was examined and shown to be statistically different (P<0.05). Results of ongoing pregnancy trials conducted at 35 days, 60 days and 90 days will be discussed. However, preliminary data indicate that individual bulls demonstrate distinctly different fertility performance in vitro. Findings from conception rates would provide a useful tool to aid selection of bovine in vitro-derived embryos for vitrification and embryo transfer in commercial settings.

Keywords: blastocyst, embryo transfer, in vitro-derived embryos, ovum pick-up, vitrification

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Authors: Jutharat Attawet, Alex Y. Wang, Cindy M. Farquhar, Elizabeth A. Sullivan

Abstract:

Background: Adverse maternal and perinatal outcomes of multiple pregnancies resulting from multiple embryo transfers (ET) has become significant concerns. This is particularly relevant for gestational carriers since they usually do not have infertility issues. Single embryo transfer (SET) therefore has been encouraged to assist reproductive technology (ART) practice in order to reduce multiple pregnancies. Objectives: This systematic review aims to investigate the pregnancy and birth outcomes of SET and multiple ET in surrogacy arrangements. Search methods: This study is a systematic review. Electronic databases were searched from CINAHL, Medline, Embase, Scopus and ProQuest for studies from 1980 to 2017. Cross-references and national ART reports were also manual searchings. Articles without restriction of English language and study types were accessed. Carrier cycles involving in SET and multiple ET were identified in database searching. The main outcome measures including clinical pregnancy, live delivery and multiple deliveries per gestational carrier cycle were compared between SET and multiple ET. Mantel-Haenzel risk ratios (RRs) with 95% confidence intervals (CIs), using the numbers of outcome events in SET and multiple ET of each study were calculated suing RevMan5.3. Outcomes: The search returned 97 articles of which 5 met the inclusion criteria. Approximately 50% of carrier cycles were transferred a single embryo and 50% were transferred more than one embryo. The clinical pregnancy rate (CPR) was 39% for SET and 53% for multiple ET, which was not significantly different with RR = 0.83 (95% CI: 0.67-1.03). The live delivery rate was 33% for SET and 57% for multiple ET which was not significantly different with RR = 0.78 (95% CI: 0.61-1.00). The multiple delivery rate per carrier was greater risks in the multiple ET carrier cycles (RR =0.4, 95% CI: 0.01-0.26). There were 104 sets of twins (including one set of twins selectively reduced from triplets to twins) and 1 set of triples in the multiple ET carrier cycle. In the SET carrier cycles, there were 2 sets of twins. Significance of the study: SET should be advocated among surrogate carriers to prevent multiple pregnancies and subsequent adverse outcomes for both carrier and baby. Surrogacy practice should be reviewed and surrogate carriers should be fully informed of the risk of adverse maternal and birth outcome of multiple pregnancies due to multiple embryo transfers.

Keywords: assisted reproduction, birth outcomes, carrier, gestational surrogacy, multiple embryo transfer, multiple pregnancy, pregnancy outcomes, single embryo transfer, surrogate mother, systematic review

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Authors: Ayman A. Swelum, Abdullah N. Al-Owaimer, Mohamed A. Abouheif

Abstract:

This study aimed to evaluate the effects of sheep breed and the number of embryos transferred on the success of multiple ovulation and embryo transfer (MOET). Sixteen Najdi and Naeimi ewes were used as donors. Multiple ovulation was achieved using equine chorionic gonadotropin (eCG). Thirty-five recipient ewes were divided into four groups: Najdi or Naeimi ewes that received either one or two embryos. After lambing, the gestation length, litter size, and sex of the lambs were recorded. The rates of pregnancy, lambing, and embryo survival were lower in the recipient Najdi than Naeimi ewes when two embryos were transferred. In contrast, the Naeimi ewes that received one embryo had a significantly lower embryo transfer success. In conclusion, the response of ewes to multiple ovulation stimulation using eCG was significantly high in Naeimi ewes (9.8±1.17). Moreover, transferring one embryo resulted in a significantly high pregnancy rate in the Najdi sheep (60%).

Keywords: embryo transfer, multiple ovulation, Najdi, Naeimi, sheep

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Authors: Sayantika Biswas, Dipanshu Sur, Amitoj Athwal, Ratnabali Chakravorty

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Title: Impact of serum estrogen and progesterone levels in the outcome pregnancy rate in frozen embryo transfer cycles. A prospective cohort study Objective: The aim of the current study was to evaluate the effect of serum estradiol (E2) and progesterone (P4) levels at different time points on pregnancy outcomes in frozen embryo transfer (FET) cycles. Materials & Method: A prospective cohort study was performed in patients undergoing frozen embryo transfer. Patients under age 37 years of age with at least one good blastocyst or three good day 3 embryos were included in the study. For endometrial preparation, 14 days of oral estradiol use (2X2 mg for 5 days. 3X2 mg for 4 days, and 4X2 mg for 5 days) was followed by vaginal progesterone twice a day and 50 mg intramuscular progesterone twice a day. Embryo transfer was scheduled 72-76 hrs or 116-120hrs after the initiation of progesterone. Serum E2 and P4 levels were examined at 4 times a) at the start of the menstrual cycle prior to the hormone supplementation. b) on the day of P4 start. c) on the day of ET. d) on the third day after ET. Result: A total 41 women were included in this study (mean age 31.8; SD 2.8). Clinical pregnancy rate was 65.55%. Serum E2 levels on at the start of the menstrual cycle prior to the hormone supplementation and on the day of P4 start were high in patients who achieved pregnancy compared to who did not (P=0.005 and P=0.019 respectively). P4 levels on on the day of ET were also high in patients with clinical pregnancy. On the day of P4 start, a serum E2 threshold of 186.4 pg/ml had a sensitivity of 82%, and P4 had a sensitivity of 71% for the prediction of clinical pregnancy at the threshold value 16.00 ng/ml. Conclusion: In women undergoing FET with hormone replacement, serum E2 level >186.4 pg/ml on the day of the start of progesterone and serum P4 levels >16.00 ng/ml on embryo transfer day are associated with clinical pregnancy.

Keywords: serum estradiol, serum progesterone, clinical pregnancy, frozen embryo transfer

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Authors: Göksel Doğan, Murat Öztürk, Didar Tuğçe Karakulak, Mehmet Nurullah Orman, Nicolas Sylvius, Matthew Blades, Mustafa Sandıkçı, Cengiz Ünsal, Mehtap Kılıç Eren, Funda Kıral, Levent Karagenç

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Assisted reproduction is associated with impaired glucose metabolism in adulthood. miRNAs are key regulators of glucose metabolism. Whether embryo culture and/or transfer alters the expression of miRNAs and to what extent this process affects glucose metabolism remain largely unknown. The purpose of the present study was to examine the expression of miRNAs in the liver in mice obtained by the transfer of blastocysts. The study was comprised of an experimental (EG) and a control group (CG). EG was generated by embryo transfer to pseudo-pregnant females. Mice born from naturally ovulating females were used as the CG. Differential expression of miRNAs, blood glucose, plasma insulin, liver glycogen, and activities of some of the rate-limiting enzymes involved in glucose metabolism were determined at ten weeks of age. Blood glucose, plasma insulin, and glycogen concentrations were similar between the groups in both sexes. Activities of enzymes were similar among females. EG males had significantly less glucokinase and phosphofructokinase activity compared to CG males. None of the miRNAs were differentially expressed in males. On the other hand, miR-143-3p expression was upregulated in EG females. Expression of none of the genes targeted by miR143-3p differed between the groups. These results demonstrate that miR143-3p, a novel regulator of type 2 diabetes, is upregulated in mice generated by assisted reproduction in a sexually-dimorphic manner with no apparent effect on glucose and insulin levels at ten weeks of age. It remains to be determined if this process is associated with impaired glucose homeostasis in the long term.

Keywords: assisted reproduction, blastocyst, embryo culture, glucose metabolism, miR143-3p, oxygen

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Authors: Hwang Kwon

Abstract:

Objective: To assess the efficacy of embryo transfer (ET) of accumulated vitrified embryos and compare pregnancy outcomes between ET of thawed embryos following accumulation of vitrified embryos (frozen ET) and ET of fresh and thawed frozen embryos following accumulation of vitrified embryos (fresh ET + frozen ET). Study design: Patients were poor ovarian responders defined according to the Bologna criteria as well as a subgroup of women whose previous IVF-ET cycle through controlled ovarian stimulation (COS) yielded one or no embryos. Sixty-four frozen ETs were performed following accumulation of vitrified embryos (ACCE )(ACCE Frozen) and 51 fresh + frozen ETs were performed following accumulation of vitrified embryos (ACCE Fresh + Frozen). Positive βhCG rate, clinical pregnancy rate, ongoing pregnancy rate, and good quality embryos (%, ±SD) were compared between two groups. Results: There were more good quality embryos in the ACCE Fresh + Frozen group than in the ACCE Frozen group: 60±34.7 versus 42.9±28.9, respectively (p=0.03). Positive βhCG rate [18/64(28.2%) vs. 13/51(25.5%); p=0.75] and clinical pregnancy rate [12/64 (18.8%) vs. 11/51 (10.9%); p=0.71] were comparable between the two groups. Conclusion: Accumulation of vitrified embryos is an effective method in patients with poor ovarian response who fulfill the Bologna criteria. Pregnancy outcomes were comparable between the two groups.

Keywords: accumulation of embryos, frozen embryo transfer, poor responder, Bologna criteria

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Authors: Mohsen Bahrami, Banafsheh Nikmehr, Yueqiang Song, Anuradha Koduru, Ayse K. Vuruskan, Hongkun Lu, Tamer M. Yalcinkaya

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In recent years, we have witnessed an explosion of studies aimed at using a combination of artificial intelligence (AI) and time-lapse imaging data on embryos to improve IVF outcomes. However, despite promising results, no study has used a matched cohort of transferred embryos which only differ in pregnancy outcome, i.e., embryos from a single clinic which are similar in parameters, such as: morphokinetic condition, patient age, and overall clinic and lab performance. Here, we used time-lapse data on embryos with known pregnancy outcomes to see if the rich spatiotemporal information embedded in this data would allow the prediction of the pregnancy outcome regardless of such critical parameters. Methodology—We did a retrospective analysis of time-lapse data from our IVF clinic utilizing Embryoscope 100% of the time for embryo culture to blastocyst stage with known clinical outcomes, including live birth vs nonpregnant (embryos with spontaneous abortion outcomes were excluded). We used time-lapse data from 200 elective single transfer embryos randomly selected from January 2019 to June 2021. Our sample included 100 embryos in each group with no significant difference in patient age (P=0.9550) and morphokinetic scores (P=0.4032). Data from all patients were combined to make a 4th order tensor, and feature extraction were subsequently carried out by a tensor decomposition methodology. The features were then used in a machine learning classifier to classify the two groups. Major Findings—The performance of the model was evaluated using 100 random subsampling cross validation (train (80%) - test (20%)). The prediction accuracy, averaged across 100 permutations, exceeded 80%. We also did a random grouping analysis, in which labels (live birth, nonpregnant) were randomly assigned to embryos, which yielded 50% accuracy. Conclusion—The high accuracy in the main analysis and the low accuracy in random grouping analysis suggest a consistent spatiotemporal pattern which is associated with pregnancy outcomes, regardless of patient age and embryo morphokinetic condition, and beyond already known parameters, such as: early cleavage or early blastulation. Despite small samples size, this ongoing analysis is the first to show the potential of AI methods in capturing the complex morphokinetic changes embedded in embryo time-lapse data, which contribute to successful pregnancy outcomes, regardless of already known parameters. The results on a larger sample size with complementary analysis on prediction of other key outcomes, such as: euploidy and aneuploidy of embryos will be presented at the meeting.

Keywords: IVF, embryo, machine learning, time-lapse imaging data

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Authors: Nathaniel Wanmi, O. M. Samuel, N. Plang, P. O. Brenda

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The wild helmeted guinea fowl has in recent time been used for research in the field of anatomy because of its peculiarity from other domesticated species of avian. Eggs of the wild helmeted guinea fowl are considered to be nutritious and has been used for medicinal purposes in some rural settlements in Nigeria. Eggs of the wild helmeted guinea fowl were purchased from hunters and taken to the National Veterinary Research Institution (NVRI) for incubation. Immediately fresh eggs were purchased, it was kindle using high powered light because of its thick egg shell and only eggs which have not started developing will be incubated and that marks the first day of incubation. On day 3 of incubation, large patches of appears redden on the surface of the egg yolk. These congested sites, develop around portion were future embryo will formed. Blood vessel were first, observed on day 4 of incubation and as days on, as embryo increases in size, blood vessels increase as well. The point of embryo implantation is evident first; by formation of congested areas and most importantly, a single zone of circular red rim. This mark the point of implantation. Blood vessels of the wild helmeted guinea fowl develops from the surface of the egg yolk, which appears initially as small strips of line. Blood vessels connects to the site of embryo implantation on day 3 of incubation. Blood vessel is the first structure to be form prior to the manifestation of the embryo.

Keywords: brain, development, helmeted, incubation

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Authors: Hasan Basri Jumin, Danil Endriand Basri

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Isolated protoplast from embryogenic callus of orange Jessamine (Murraya paniculata L. (Jack) cultured and maintained under growth chamber at the temperature +25oC. The parameter observed are the plating efficiency, the number of spherical embryos, heard-shaped embryos-like structure, shoot formation, and plantlets obtained. Treatment was arranged with 0.0, 0.001, 0.01, 0.1 or 1.0 mg 1-1 Naphthalene acetic acid (NAA), and 0, 300, 500 mg 1/l malt extract (ME) and 0.M sorbitol in the medium with 2.5 % sucrose. Interaction between 0.001 mg/l NAA and 500 mg/l was observed the higher percentage of planting efficiency. For embryo development from callus, the media was added to 0.0 mg/l, 0.001 mg/l, 0.01 ,mg/l, 0.1 mg/l, 1.0 mg/l NAA, and 1.0 %, 2.0 %, 3.0 %, 4.0 % sucrose. Media supplemented with 0.01mg/l NAA, and 1.0% sucrose was found to be a suitable medium for the development of spherical somatic embryos. A combination of 0.1 mg/ indole acetic acid (IAA) and 0.1 mg/l zeatin constituted the spherical somatic embryo became heart-shaped embryos-like structure. A combination between GA3 0.1 mg 1/l GA3 and 0.1 mg 1-1 zeatin is looking high, growing the heart-shaped embryos-like structure to form a shoot. Cells were developed into spherical embryos and grew into heart-shaped embryos, and then spherical somatic embryos developed into shoot formation. Sequence from single protoplast to plantlets was obtained by using a low concentration of plant growth regulator and sucrose; This recovery of single protoplast to be completed plantlets is a new technology in plant cell culture, and this could be used in genetic engineering in citrus.

Keywords: heart-shaped-embryos-like-structure, Muraya-paniculata, plant-growth-regulator, spherical- somatic-embryo, single protoplast, glucose

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Authors: Huajuan Shi

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Background: The biopsy of the preimplantation embryo may increase the potential risk and concern of embryo viability. Clinically discarded spent embryo medium (SEM) has entered the view of researchers, sparking an interest in noninvasive embryo screening. However, one of the major restrictions is the extremelty low quantity of cf-RNA, which is difficult to efficiently and unbiased amplify cf-RNA using traditional methods. Hence, there is urgently need to an efficient and low bias amplification method which can comprehensively and accurately obtain cf-RNA information to truly reveal the state of SEM cf-RNA. Result: In this present study, we established an agarose PCR amplification system, and has significantly improved the amplification sensitivity and efficiency by ~90 fold and 9.29 %, respectively. We applied agarose to sequencing library preparation (named AG-seq) to quantify and characterize cf-RNA in SEM. The number of detected cf-RNAs (3533 vs 598) and coverage of 3' end were significantly increased, and the noise of low abundance gene detection was reduced. The increasing percentage 5' end adenine and alternative splicing (AS) events of short fragments (< 400 bp) were discovered by AG-seq. Further, the profiles and characterizations of cf-RNA in spent cleavage medium (SCM) and spent blastocyst medium (SBM) indicated that 4‐mer end motifs of cf-RNA fragments could remarkably differentiate different embryo development stages. Significance: This study established an efficient and low-cost SEM amplification and library preparation method. Not only that, we successfully described the characterizations of SEM cf-RNA of preimplantation embryo by using AG-seq, including abundance features fragment lengths. AG-seq facilitates the study of cf-RNA as a noninvasive embryo screening biomarker and opens up potential clinical utilities of trace samples.

Keywords: cell-free RNA, agarose, spent embryo medium, RNA sequencing, non-invasive detection

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Authors: Alexander V. Spirov, David M. Holloway, Ekaterina M. Myasnikova

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Segmentation of the early Drosophila embryo results from the dynamic establishment of spatial gene expression patterns. Patterning occurs on an embryo geometry which is a 'deformed' prolate ellipsoid, with anteroposterior and dorsal-ventral major and minor axes, respectively. Patterning is largely independent along each axis, but some interaction can be seen in the 'bending' of the segmental expression stripes. This interaction is not well understood. In this report, we investigate how 3D geometrical features of the early embryo affect the segmental expression patterning. Specifically, we study the effect of geometry on formation of the Bicoid primary morphogenetic gradient. Our computational results demonstrate that embryos with a much longer ventral than dorsal surface ('bellied') can produce curved Bicoid concentration contours which could activate curved stripes in the downstream pair-rule segmentation genes. In addition, we show that having an extended source for Bicoid in the anterior of the embryo may be necessary for producing the observed exponential form of the Bicoid gradient along the anteroposterior axis.

Keywords: Drosophila embryo, bicoid morphogenetic gradient, exponential expression profile, expression surface form, segmentation genes, 3D modelling

Procedia PDF Downloads 236

Authors: Huajuan Shi

Abstract:

Background: The biopsy of the preimplantation embryo may increase the potential risk and concern of embryo viability. Clinically discarded spent embryo medium (SEM) has entered the view of researchers, sparking an interest in noninvasive embryo screening. However, one of the major restrictions is the extremelty low quantity of cf-RNA, which is difficult to efficiently and unbiased amplify cf-RNA using traditional methods. Hence, there is urgently need to an efficient and low bias amplification method which can comprehensively and accurately obtain cf-RNA information to truly reveal the state of SEM cf-RNA. Result: In this present study, we established an agarose PCR amplification system, and has significantly improved the amplification sensitivity and efficiency by ~90 fold and 9.29 %, respectively. We applied agarose to sequencing library preparation (named AG-seq) to quantify and characterize cf-RNA in SEM. The number of detected cf-RNAs (3533 vs 598) and coverage of 3' end were significantly increased, and the noise of low abundance gene detection was reduced. The increasing percentage 5' end adenine and alternative splicing (AS) events of short fragments (< 400 bp) were discovered by AG-seq. Further, the profiles and characterizations of cf-RNA in spent cleavage medium (SCM) and spent blastocyst medium (SBM) indicated that 4‐mer end motifs of cf-RNA fragments could remarkably differentiate different embryo development stages. Significance: This study established an efficient and low-cost SEM amplification and library preparation method. Not only that, we successfully described the characterizations of SEM cf-RNA of preimplantation embryo by using AG-seq, including abundance features fragment lengths. AG-seq facilitates the study of cf-RNA as a noninvasive embryo screening biomarker and opens up potential clinical utilities of trace samples.

Keywords: cell-free RNA, agarose, spent embryo medium, RNA sequencing, non-invasive detection

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Authors: F. A. Hamad, S. He

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In this paper, the average heat transfer characteristics for a cross flow cylinder of 16 mm diameter in a vertical pipe has been studied for single-phase flow (water/oil) and multicomponent (non-boiling) flow (water-air, water-oil, oil-air and water-oil-air). The cylinder is uniformly heated by electrical heater placed at the centre of the element. The results show that the values of average heat transfer coefficients for water are around four times the values for oil flow. Introducing air as a second phase with water has very little effect on heat transfer rate, while the heat transfer increased by 70% in case of oil. For water–oil flow, the heat transfer coefficient values are reflecting the percentage of water up to 50%, but increasing the water more than 50% leads to a sharp increase in the heat transfer coefficients to become close to the values of pure water. The enhancement of heat transfer by mixing two phases may be attributed to the changes in flow structure near to cylinder surface which lead to thinner boundary layer and higher turbulence. For three-phase flow, the heat transfer coefficients for all cases fall within the limit of single-phase flow of water and oil and are very close to pure water values. The net effect of the turbulence augmentation due to the introduction of air and the attenuation due to the introduction of oil leads to a thinner boundary layer of oil over the cylinder surface covered by a mixture of water and air bubbles.

Keywords: circular cylinder, cross flow, hear transfer, multicomponent multiphase flow

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Authors: Banafsheh Nikmehr, Mohsen Bahrami, Yueqiang Song, Anuradha Koduru, Ayse K. Vuruskan, Hongkun Lu, Mallory Pitts, Tolga B. Mesen, Tamer M. Yalcinkaya

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The number of people who receive in vitro fertilization (IVF) treatment has increased on a startling trajectory over the past two decades. Despite advances in this field, particularly the introduction of intracytoplasmic sperm injection (ICSI) and the preimplantation genetic screening (PGS), the IVF success remains low. A major factor contributing to IVF failure is embryonic aneuploidy (abnormal chromosome content), which often results in miscarriage and birth defects. Although PGS is often used as the standard diagnostic tool to identify aneuploid embryos, it is an invasive approach that could affect the embryo development, and yet inaccessible to many patients due its high costs. As such, there is a clear need for a non-invasive cost-effective approach to identify euploid embryos for single embryo transfer (SET). The reported differences between morphokinetic behaviors of aneuploid and euploid embryos has shown promise to address this need. However, current literature is inconclusive and further research is urgently needed to translate current findings into clinical diagnostics. In this ongoing study, we found significant differences between morphokinetic behaviors of euploid and aneuploid embryos that provides important insights and reaffirms the promise of such behaviors for developing non-invasive methodologies. Methodology—A total of 242 embryos (euploid: 149, aneuploid: 93) from 74 patients who underwent IVF treatment in Carolinas Fertility Clinics in Winston-Salem, NC, were analyzed. All embryos were incubated in an EmbryoScope incubator. The patients were randomly selected from January 2019 to June 2021 with most patients having both euploid and aneuploid embryos. All embryos reached the blastocyst stage and had known PGS outcomes. The ploidy assessment was done by a third-party testing laboratory on day 5-7 embryo biopsies. The morphokinetic variables of each embryo were measured by the EmbryoViewer software (Uniesense FertiliTech) on time-lapse images using 7 focal depths. We compared the time to: pronuclei fading (tPNf), division to 2,3,…,9 cells (t2, t3,…,t9), start of embryo compaction (tSC), Morula formation (tM), start of blastocyst formation (tSC), blastocyst formation (tB), and blastocyst expansion (tEB), as well as intervals between them (e.g., c23 = t3 – t2). We used a mixed regression method for our statistical analyses to account for the correlation between multiple embryos per patient. Major Findings— The average age of the patients was 35.04 yrs. The average patient age associated with euploid and aneuploid embryos was not different (P = 0.6454). We found a significant difference in c45 = t5-t4 (P = 0.0298). Our results indicated this interval on average lasts significantly longer for aneuploid embryos - c45(aneuploid) = 11.93hr vs c45(euploid) = 7.97hr. In a separate analysis limited to embryos from the same patients (patients = 47, total embryos=200, euploid=112, aneuploid=88), we obtained the same results (P = 0.0316). The statistical power for this analysis exceeded 87%. No other variable was different between the two groups. Conclusion— Our results demonstrate the importance of morphokinetic variables as potential biomarkers that could aid in non-invasively characterizing euploid and aneuploid embryos. We seek to study a larger population of embryos and incorporate the embryo quality in future studies.

Keywords: IVF, embryo, euploidy, aneuploidy, morphokinteic

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Authors: Myung-Gon Yoon, Jung-Ho Moon, Tae Kwon Ha

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This paper considers a robust stabilization problem of a single agent in a multi-agent consensus system composed of identical agents, when the network topology of the system is completely unknown. It is shown that the transfer function of an agent in a consensus system can be described as a multiplicative perturbation of the isolated agent transfer function in frequency domain. Applying known robust stabilization results, we present sufficient conditions for a robust stabilization of an agent against unknown network topology.

Keywords: single agent control, multi-agent system, transfer function, graph angle

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Authors: Katalin Gombos, Bence Gálik, Krisztina Ildikó Kalács, Krisztina Gödöny, Ákos Várnagy, József Bódis, Attila Gyenesei, Gábor L. Kovács

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Although non-invasive pre-implantation genetic testing for aneuploidy (NIPGT-A) is potentially appropriate to assess chromosomal ploidy of the embryo, practical application of it in a routine IVF center has not been started in the absence of a recommendation. We developed a comprehensive workflow for a clinically applicable strategy for NIPGT-A based on next-generation sequencing (NGS) technology. We performed MALBAC whole genome amplification and NGS on spent blastocyst culture media of Day 3 embryos fertilized with intra-cytoplasmic sperm injection (ICSI). Spent embryonic culture media of morphologically good quality score embryos were enrolled in further analysis with the blank culture media as background control. Chromosomal abnormalities were identified by an optimized bioinformatics pipeline applying a copy number variation (CNV) detecting algorithm. We demonstrate a comprehensive workflow covering both wet- and dry-lab procedures supporting a clinically applicable strategy for NIPGT-A. It can be carried out within 48 h which is critical for the same-cycle blastocyst transfer, but also suitable for “freeze all” and “elective frozen embryo” strategies. The described integrated approach of non-invasive evaluation of embryonic DNA content of the culture media can potentially supplement existing pre-implantation genetic screening methods.

Keywords: next generation sequencing, in vitro fertilization, embryo assessment, non-invasive pre-implantation genetic testing

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Authors: Münüre Tanur Erkoyuncu, Mustafa Yorgancılar

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Crop improvement through genetic engineering depends on effective and reproducible plant regeneration systems. Immature embryos are the most widely used explant source for in vitro regeneration in barley (Hordeum vulgare L.). However, immature embryos require the continuous growth of donor plants and the suitable stage for their culture is also certainly limited. On the other hand, mature embryos can be procured and stored easily; they can be studied throughout the year. In this study, an effective callus induction and plant regeneration were aimed to develop from mature embryos of different barley genotypes. The effect of medium (MS₁ and MS₂), auxin type (2,4-D, dicamba, picloram and 2,4,5-T) and concentrations (2, 4, 6 mg/l) on callus formation and effect of cytokinin type (TDZ, BAP) and concentrations (0.2, 0.5, 1.0 mg/l) on green plant regeneration were evaluated in mature embryo culture of barley. Callus and shoot formation was successful for all genotypes. By depending on genotype, MS₁ is the best medium, 4 mg/l dicamba is the best growth regulator in the callus induction and MS₁ is the best medium, 1 mg/l BAP is the best growth regulator in the shoot formation were determined.

Keywords: barley, callus, embryo culture, mature embryo

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Authors: Surendra Bodakhe

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In this investigation, anticataract activity was determined using cataract formation in developing chick embryo by hydrocortisone. Lenses were evaluated firstly for the extent of opacity and secondly, for lens glutathione (GSH) levels. Betulinic acid was isolated from the chloroform fraction of the crude ethanolic extract of Bauhinia variegata bark (SBE). Fourteen days old Australorp fertilized eggs were divided into different groups of six eggs each. After 24 hrs incubation in a humidified incubator (37οC), at 15 days of age; hydrocortisone (0.25µM/0.2ml/egg) was administered to the chorioallantoic membrane of chick embryos through a small hole in the egg shell on the air sack. Ascorbic acid (standard) or Betulinic acid (test) were administered at 3, 10 and 20 hr after hydrocortisone administration at a specified dose. The puncture was sealed with a cellophane tape and eggs were incubated for 48 hrs in a humidified incubator at 37οC. After 48 hrs, the lenses were isolated for the determination of the extent of opacity and Glutathione level. The betulinic acid prevented the opacification of the chick embryo lenses induced by hydrocortisone. The betulinic acid also prevented the decline of GSH content caused by hydrocortisone. The results indicate that betulinic acid protect the cataract formation in chick embryo lenses induced by hydrocortisone.

Keywords: betulinic acid, cataract, cloudiness, ovine

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Authors: Jayeon Kim, Eunjung Yu, Taeki Yoon

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Most spontaneous miscarriage is believed to be a consequence of embryo aneuploidies. Transferring eukaryotic embryos selected by PGS is expected to decrease the miscarriage rate. Current PGS indications include advanced maternal age, recurrent pregnancy loss, repeated implantation failure. Recently, use of PGS for healthy women without above indications for the purpose of improving in vitro fertilization (IVF) outcomes is on the rise. However, it is still controversy about the beneficial effect of PGS in this population, especially, in women with a history of no more than 2 miscarriages or miscarriage of eukaryotic abortus. This study aimed to investigate if karyotyping result of abortus is a good indicator of preimplantation genetic screening (PGS) in subsequent IVF cycle in women with a history of spontaneous abortion. A single-center retrospective cohort study was performed. Women who had spontaneous abortion(s) (less than 3) and dilatation and evacuation, and subsequent IVF from January 2016 to November 2016 were included. Their medical information was extracted from the charts. Clinical pregnancy was defined as presence of a gestational sac with fetal heart beat detected on ultrasound in week 7. Statistical analysis was performed using SPSS software. Total 234 women were included. 121 out of 234 (51.7%) underwent karyotyping of the abortus, and 113 did not have the abortus karyotyped. Embryo biopsy was performed on 3 or 5 days after oocyte retrieval, followed by embryo transfer (ET) on a fresh or frozen cycle. The biopsied materials were subjected to microarray comparative genomic hybridization. Clinical pregnancy rate per ET was compared between PGS and non-PGS group in each study group. Patients were grouped by two criteria: karyotype of the abortus from previous miscarriage (unknown fetal karyotype (n=89, Group 1), eukaryotic abortus (n=36, Group 2) or aneuploidy abortus (n=67, Group 3)), and pursuing PGS in subsequent IVF cycle (pursuing PGS (PGS group, n=105) or not pursuing PGS (non-PGS group, n=87)). The PGS group was significantly older and had higher number of retrieved oocytes and prior miscarriages compared to non-PGS group. There were no differences in BMI and AMH level between those two groups. In PGS group, the mean number of transferable embryos (eukaryotic embryo) was 1.3 ± 0.7, 1.5 ± 0.5 and 1.4 ± 0.5, respectively (p = 0.049). In 42 cases, ET was cancelled because all embryos biopsied turned out to be abnormal. In all three groups (group 1, 2, and 3), clinical pregnancy rates were not statistically different between PGS and non-PGS group (Group 1: 48.8% vs. 52.2% (p=0.858), Group 2: 70% vs. 73.1% (p=0.730), Group 3: 42.3% vs. 46.7% (p=0.640), in PGS and non-PGS group, respectively). In both groups who had miscarriage with eukaryotic and aneuploidy abortus, the clinical pregnancy rate between IVF cycles with and without PGS was not different. When we compare miscarriage and ongoing pregnancy rate, there were no significant differences between PGS and non-PGS group in all three groups. Our results show that the routine application of PGS in women who had less than 3 miscarriages would not be beneficial, even in cases that previous miscarriage had been caused by fetal aneuploidy.

Keywords: preimplantation genetic diagnosis, miscarriage, kpryotyping, in vitro fertilization

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Authors: Rita Emília Szabó, Róbert Polanek, Tünde Tőkés, Zoltán Szabó, Szabolcs Czifrus, Katalin Hideghéty

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Purpose: Several feature of zebrafish are making them amenable for investigation on therapeutic approaches such as ionizing radiation. The establishment of zebrafish model for comprehensive radiobiological research stands in the focus of our investigation, comparing the radiation effect curves of neutron and photon irradiation. Our final aim is to develop an appropriate vertebrate model in order to investigate the relative biological effectiveness of laser driven ionizing radiation. Methods and Materials: After careful dosimetry series of viable zebrafish embryos were exposed to a single fraction whole-body neutron-irradiation (1,25; 1,875; 2; 2,5 Gy) at the research reactor of the Technical University of Budapest and to conventional 6 MeV photon beam at 24 hour post-fertilization (hpf). The survival and morphologic abnormalities (pericardial edema, spine curvature) of each embryo were assessed for each experiment at 24-hour intervals from the point of fertilization up to 168 hpf (defining the dose lethal for 50% (LD50)). Results: In the zebrafish embryo model LD50 at 20 Gy dose level was defined and the same lethality were found at 2 Gy dose from the reactor neutron beam resulting RBE of 10. Dose-dependent organ perturbations were detected on macroscopic (shortening of the body length, spine curvature, microcephaly, micro-ophthalmia, micrognathia, pericardial edema, and inhibition of yolk sac resorption) and microscopic (marked cellular changes in skin, cardiac, gastrointestinal system) with the same magnitude of dose difference. Conclusion: In our observations, we found that zebrafish embryo model can be used for investigating the effects of different type of ionizing radiation and this system proved to be highly efficient vertebrate model for preclinical examinations.

Keywords: ionizing radiation, LD50, relative biological effectiveness, zebrafish embryo

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Authors: Sri Wahjuningsih, Nur Ihsan, Hadiah

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In the embryo transfer program, to address the limited production of embryos in vivo, in vitro embryo production has become an alternative approach that is relatively inexpensive. One potential source of embryos that can be developed is to use immature oocytes then conducted in vitro maturation and in vitro fertilization. However, obstacles encountered were oocyte viability mammals have very limited that it cannot be stored for a long time, so we need oocyte cryopreservation. The research was conducted to know the optimal concentration use of ethylene glycol as a cryoprotectant on oocytes freezing.Material use in this research was immature oocytes; taken from abbatoir which was aspirated from follicle with diameter 2-6 mm. Concentration ethylen glycol used were 0,5 M, I M, 1,5 M and 2M. The freezing method used was conventional method combined with a five-step protocol washing oocytes from cryoprotectant after thawing. The result showed that concentration ethylen glycol have the significant effect (P<0.05) on oocytes quality after thawing and in vitro maturation. It was concluded that concentration 1,5 M was the best concentration for freezing oocytes using conventional method.

Keywords: bovine, conventional freezing, ethylen glycol, oocytes

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Authors: Vasudev Das

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The specific problem is that many leaders of the 21st century do not understand that the extermination of embryos wreaks havoc on peace leadership. The purpose of the document review is to understand embryology in facilitating peace leadership. Extermination of human embryos generates a requital wave of violence which later falls on human society in the form of disturbances, considering that violence breeds further violence as a consequentiality. The study results reveal that a deep understanding of embryology facilitates peace leadership, given that minimizing embryo extermination enhances non-violence in the global village. Neo-Newtonians subscribe to the idea that every action has an equal and opposite reaction. The US Federal Government recognizes the embryo or fetus as a member of Homo sapiens. The social change implications of this study are that understanding human embryology promotes peace leadership, considering that the consequentiality of embryo extermination can serve as a deterrent for violence on embryos.

Keywords: consequentiality, Homo sapiens, neo-Newtonians, violence

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Authors: G. Ravindranath, S. Savitha

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This paper presents heat transfer analysis of single horizontal bare tube and heat transfer analysis of staggered arrangement of bare tube bundles bare tube bundles in gas-solid (air-solid) fluidized bed and predictions are done by using Artificial Neural Network (ANN) based on experimental data. Fluidized bed provide nearly isothermal environment with high heat transfer rate to submerged objects i.e. due to through mixing and large contact area between the gas and the particle, a fully fluidized bed has little temperature variation and gas leaves at a temperature which is close to that of the bed. Measurement of average heat transfer coefficient was made by local thermal simulation technique in a cold bubbling air-fluidized bed of size 0.305 m. x 0.305 m. Studies were conducted for single horizontal Bare Tube of length 305mm and 28.6mm outer diameter and for bare tube bundles of staggered arrangement using beds of large (average particle diameter greater than 1 mm) particle (raagi and mustard). Within the range of experimental conditions influence of bed particle diameter ( Dp), Fluidizing Velocity (U) were studied, which are significant parameters affecting heat transfer. Artificial Neural Networks (ANNs) have been receiving an increasing attention for simulating engineering systems due to some interesting characteristics such as learning capability, fault tolerance, and non-linearity. Here, feed-forward architecture and trained by back-propagation technique is adopted to predict heat transfer analysis found from experimental results. The ANN is designed to suit the present system which has 3 inputs and 2 out puts. The network predictions are found to be in very good agreement with the experimental observed values of bare heat transfer coefficient (hb) and nusselt number of bare tube (Nub).

Keywords: fluidized bed, large particles, particle diameter, ANN

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Authors: Irianes C. Gozali, Betty J.L. Laglbauer, Muhammad G. Salim, Sila K. Sari, Fahmi Fahmi, Selvia Oktaviyani

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Muncar, located off the eastern coast of Java, is an important fishing port for small-scale fleets which land mobulid rays as retained bycatch, primarily in drift gillnets. Due to overlap with fishing grounds in the Bali Strait, three devil ray species are landed in Muncar, the spinetail devil ray Mobula mobular, the bentfin devil ray Mobula thurstoni, and the Chilean devil ray Mobula tarapacana, which are all listed as Endangered by the International Union for the Conservation of Nature. However, despite the importance of life-history data to better manage stocks, such information is still rare or unavailable for Indonesian mobulid ray populations. Using morphometric data, reproductive assessments, and samples collected from dead specimens at fish markets from 2015-2019, we provide information on the maturity stage, reproductive periodicity, gestation, and size at parturition. A majority of immature individuals of all three devil ray species were recorded (<10% individuals in Mobula mobular to <30% individuals in Mobula thurstoni). Pregnant females of two species, Mobula mobular and Mobula thurstoni were recorded containing embryos of various developmental stages (each with a single embryo in the left functional uterus), while for Mobula tarapacana, no fetuses were found. The largest embryo recorded in M. mobular was within the range of that previously reported for neonates of the species in Indonesia (957 cm, for a 920-994 range), and represents a near-term embryo reflecting size at parturition. Low reproductive output was confirmed for the study-species. Based on this study, we infer that the Bali Straight is likely an important location for devil ray reproduction, which raises concern for the sustainability of mobulid ray populations in the face of bycatch in drift gillnets. Potential management approaches to tackle this issue are discussed.

Keywords: devil ray, mobulid, reproduction, Indonesia

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Authors: Hao Xiaohong, Wu Zongxiang, Chen Xuefeng

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With the high power density and high integration of electronic devices, their heat flux has been increasing rapidly. Therefore, an effective cooling technology is essential for the reliability and efficient operation of electronic devices. Liquid cooling is studied increasingly widely for its higher heat transfer efficiency. Serpentine channels are superior in the augmentation of single-phase convective heat transfer because of their better channel velocity distribution. In this paper, eight different frame sizes water-cooled serpentine channel heat sinks are designed to study the heat transfer and pressure drop characteristics. With water as the working fluid, experiment setup is established and the results showed the effect of different channel width, fin thickness and number of channels on thermal resistance and pressure drop.

Keywords: heat transfer, experiment, serpentine heat sink, pressure drop

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Authors: Du Yanxia, Stephan E. Bansmer, Gui Yewei, Xiao Guangming, Yang Xiaofeng

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The heat and mass transfer characteristics of icing coupled with film flow is studied and the coupled model of the thermal behavior with the flow simulation by single-step method is developed. The behavior of ice and water was analyzed. The results show that under supercooled large droplet (SLD) icing conditions, the film flow is an important phonomena in icing accretion process. The pressure gradient, gravity and shear stress are the main factors affecting the film flow on icing surface, which has important influence on the shape and rate of icing. To predict SLD ice accretion accurately, the heat and mass transfer of ice and film flow should be taken into account.

Keywords: SLD, aircraft, icing, heat and mass transfer

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Authors: I. Bochev, A. Shterev, S. Kyurkchiev

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One of the factors associated with poor success rates in human in vitro fertilization (IVF) is the suboptimal culture conditions in which fertilization and early embryonic growth occur. Co-culture systems with helper cell lines appear to enhance the in vitro conditions and allow embryos to demonstrate improved in vitro development. The co-culture of human embryos with monolayers of autologous endometrial stromal cell (EnSCs) results in increased blastocyst development with a larger number of blastomeres, lower incidence of fragmentation and higher pregnancy rates in patients with recurrent implantation failure (RIF). The aim of the study was to examine the influence of autologous endometrial stromal cell (EnSC) co-culture on day 3 embryo quality by comparing the morphological status of the embryos from the same patients undergoing consecutive IVF/Intracytoplasmic sperm injection (ICSI) cycles without and with EnSC co-culture. This retrospective randomized study (2015-2017) includes 20 couples and a total of 46 IVF/ICSI cycles. Each patient couple included had at least two IVF/ICSI procedures – one with and one without autologous EnSC co-culture. Embryo quality was assessed at 68±1 hours in culture, according to Istanbul consensus criteria (2010). Day 3 embryos were classified into three groups: good – grade 1; fair – grade 2; poor – grade 3. Embryos from all cycles were divided into two groups (A – co-cultivated; B – not co-cultivated) and analyzed. Second, for each patient couple, embryos from matched IVF/ICSI cycles (with and without co-culture) were analyzed separately. When an analysis of co-cultivated day 3 embryos from all cycles was performed (n=137; group A), 43.1% of the embryos were graded as “good”, which was not significantly different from the respective embryo quality rate of 42.2% (p = NS) in group B (n=147) with non-co-cultivated embryos. The proportions of fair and poor quality embryos in group A and group B were similar as well – 11.7% vs 10.2% and 45.2% vs 47.6% (p=NS), respectively. Nevertheless, the separate embryo analysis by matched cycles for each couple revealed that in 65% of the cases the proportion of morphologically better embryos was increased in cycles with co-culture in comparison with those without co-culture. A decrease in this proportion after endometrial stromal cell co-cultivation was found in 30% of the cases, whereas no difference was observed in only one couple. The results demonstrated that there is no marked difference in the overall morphological quality between co-cultured and non-co-cultured embryos on day 3. However, in significantly greater percentage of couples the process of autologous EnSC co-culture could increase the proportion of morphologically improved day 3 embryos. By mimicking the in vivo relationship between embryo and maternal environment, co-culture in autologous EnSC system represents a perspective approach to improve the quality of embryos in cases with elevated risk for development of embryos with impaired morphology.

Keywords: autologous endometrial stromal cells, co-culture, day 3 embryo, morphological quality

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Authors: A. Falavand Jozaei, A. Ghafouri, M. Mosavi Navaei

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In this paper for a given heat duty, study of number of baffles on pressure drop and heat transfer is considered in a STHX (Shell and Tube Heat Exchanger) with single segmental baffles. The effect of number of baffles from 9 to 52 baffles (baffle spacing variations from 4 to 24 inches) over OHTC (Overall Heat Hransfer Coefficient) to pressure drop ratio (U/Δp ratio). The results show that U/Δp ratio is low when baffle spacing is minimum (4 inches) because pressure drop is high; however, heat transfer coefficient is very significant. Then, with the increase of baffle spacing, pressure drop rapidly decreases and OHTC also decreases, but the decrease of OHTC is lower than pressure drop, so (U/Δp) ratio increases. After increasing baffles more than 12 inches, variation in pressure drop is gradual and approximately constant and OHTC decreases; Consequently, U/Δp ratio decreases again. If baffle spacing reaches to 24 inches, STHX will have minimum pressure drop, but OHTC decreases, so required heat transfer surface increases and U/Δp ratio decreases. After baffle spacing more than 12 inches, variation of shell side pressure drop is negligible. So optimum baffle spacing is suggested between 8 to 12 inches (43 to 63 percent of inside shell diameter) for a sufficient heat duty and low pressure drop.

Keywords: shell and tube heat exchanger, single segmental baffle, overall heat transfer coefficient, pressure drop

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Authors: Abhishek Kumar, Mohamed Awadein, Georg Gramse, Luyang Song, He Sun, Wolfgang Schofberger, Stefan Müllegger

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Electron transfer is a crucial part of chemical reactions which drive everyday processes. With the help of an electro-chemical radio frequency scanning tunneling microscopy (EC-RF-STM) setup, we are observing single electron mediated oxidation-reduction processes in molecules like ferrocene and transition metal corroles. Combining the techniques of scanning microwave microscopy and cyclic voltammetry allows us to monitor such processes with attoampere sensitivity. A systematic study of such phenomena would be critical to understanding the nano-scale behavior of catalysts, molecular sensors, and batteries relevant to the development of novel material and energy applications.

Keywords: radiofrequency, STM, cyclic voltammetry, ferrocene

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Authors: Eun Jeong Yu, Inn Soo Kang, Tae Ki Yoon, Mi Kyoung Koong

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Advanced female age is associated with higher cycle cancelation rates, lower clinical pregnancy rate, increased miscarriage and aneuploidy rates in IVF (In Vitro Fertilization) cycles. This retrospective cohort study was conducted at a Cha Fertility Center, Seoul Station. All fresh non-donor IVF cycles performed in women aged 40 years and above from January 2016 to December 2016 were reviewed. Donor/recipient treatment, PGD/PGS (Preimplantation Genetic Diagnosis/ Preimplantation Genetic Screening) were excluded from analysis. Of the 1,166 cycles from 753 women who completed ovulation induction, 1,047 were appropriate for the evaluation according to inclusion and exclusion criterion. IVF cycles were categorized according to age and grouped into the following 1-year age groups: 40, 41, 42, 43, 44, 45 and > 46. The mean age of patients was 42.4 ± 1.8 years. The median AMH (Anti-Mullerian Hormone) level was 1.2 ± 1.5 ng/mL. The mean number of retrieved oocytes was 4.9 ± 4.3. The clinical pregnancy rate and live birth rate in women > 40 years significantly decreased with each year of advancing age (p < 0.001). The clinical pregnancy rate decreased from 21% at the age of 40 years to 0% at ages above 45 years. Live birth rate decreased from 12.3% to 0%, respectively. There were no clinical pregnancy outcomes among 95 patients aged above 45 years of age. The overall miscarriage rate was 40.7% (range, 36.7%-70%). The transfer of at least one good quality embryo was associated with about 4-9% increased chance of a clinical pregnancy rate. Therefore, IVF in old age women less than 46 had a reasonable chance for successful pregnancy outcomes especially when good quality embryo is transferred.

Keywords: advanced maternal age, in vitro fertilization, pregnancy rate, live birth rate

Procedia PDF Downloads 112