198 Comparison of invitro production of bison and cattle embryos and effect of L-carnitine during maturation of bison oocytes

2020 ◽  
Vol 32 (2) ◽  
pp. 227
Author(s):  
A. R. Moawad ◽  
H. Benham ◽  
J. P. Barfield
Keyword(s):  
Assisted Reproductive Technologies ◽  
Cell Mass ◽  
Reproductive Technologies ◽  
T Test ◽  
Control Group ◽  
Chi Square ◽  
Important Species ◽  
Developmental Potential ◽  
Cell Numbers ◽  
Frozen Semen

Bison are an important species in North America, both economically and culturally. Although assisted reproductive technologies have been applied to preserve the genetic diversity of bison, development of these technologies remains limited for this species. The objective of the present study was to compare success rates of oocyte maturation, fertilization, and embryo development invitro in bison versus cattle (experiment 1). Cumulus-oocyte complexes obtained from abattoir-derived cattle and bison ovaries were matured, fertilized with frozen semen, and cultured invitro using standard procedures (De La Torre-Sanchez et al. 2006 Reprod. Fertil. Dev. 18, 585-596). At least three replicates were repeated for each experimental group. Oocyte recovery rate was lower in bison than in cattle (4.3; 2797/699 vs. 6.7; 4138/677, oocyte/ovary; P<0.01, t-test). Nuclear maturation (oocytes at MII, 23h post-IVM) and fertilization rates (oocytes with 2 pronuclei 18h post-insemination; p.i.) evaluated by Hoechst stain were lower (P<0.01, chi-square) for bison (65.1%; 56/86 and 32.7%; 18/55, respectively) than for cattle (88.3%; 83/94 and 70.9%; 39/55, respectively). Polyspermy tended to be higher in bison than in cattle (12.7% vs. 3.6%, P=0.08). The percentages of 2-cell embryos tended to be lower in bison than in cattle (13.5% vs. 25.0%, P>0.05) at 24h p.i. but by 30h p.i., this difference increased (33.7% vs. 67.0%, P<0.01, chi-square). Cleavage (Day 3) and blastocyst (Day 7) rates were lower (P<0.01, chi-square) for bison (58.2%; 280/481 and 14.6%; 70/481, respectively) than for cattle (90.8%; 405/446 and 22.9%; 102/446, respectively). Total cell number (74.9±4.8 vs.114.2±5.8), trophectoderm cell numbers (57.9±4.6 vs. 89.2±4.8) and inner cell mass cell numbers (16.9±2.3 vs. 25±1.9) as determined by Hoechst and propidium iodide were all lower (P<0.01, t-test) in bison than in cattle blastocysts. To improve oocyte competence in bison, we evaluated effects of L-carnitine (LC) supplementation during IVM on developmental potential of bison oocytes (experiment 2). Cumulus-oocyte complexes were matured in IVM medium supplemented with 0, 0.15, 0.3, 0.6, or 1.2mgmL−1 LC. No differences were observed in cleavage rates of control (0mgmL−1 LC) and LC-treated groups (values ranged from 60.0 to 66.4%). Interestingly, a dose-dependent increase in blastocyst development was found with the lowest value recorded in control group (10.4%; 14/134) and the highest value in the 1.2mgmL−1 LC supplemented group (22.2%; 23/105; P<0.01, chi-square, n=4). Adding 1.2mgmL−1 LC to the IVM medium improved the percentage of hatching blastocysts compared with the control. In conclusion, bison oocytes exhibited lower invitro maturation, fertilization, and developmental rates compared with cattle oocytes using our system, and bison embryos were delayed in the timing of first cleavage. L-Carnitine supplementation during IVM of bison oocytes improved the preimplantation development and quality of invitro-produced blastocysts.

132 TIMING OF THE FIRST ZYGOTIC CLEAVAGE WAS NOT RELATED TO THE SHIFT IN SEX RATIO OF BOVINE BLASTOCYST IN VITRO

2009 ◽  
Vol 21 (1) ◽  
pp. 165
Author(s):  
K. Zaorska ◽  
E. Pers-Kamczyc ◽  
D. Lechniak
Keyword(s):  
Sex Ratio ◽  
Positive Influence ◽  
Control Group ◽  
Fish Analysis ◽  
Chi Square ◽  
Experimental Conditions ◽  
Female Ratio ◽  
Developmental Potential ◽  
Frozen Semen

It has been shown that embryos which cleaved earlier (<30 h postinsemination, hpi) have greater chance to develop to blastocyst. It has been shown that in in vitro conditions male embryos develop faster than females. Further, spermatozoa with the Y chromosome more frequently penetrate oocytes during the first 6 hpi. The aim of this experiment was to investigate the sex-related embryo growth rate in relation to the timing of the first zygotic cleavage and GH presence during IVM. Oocytes were matured in TCM-199 supplemented with fatty acid free BSA and hormones (FSH and GH) and then inseminated (Parrish et al. 1998 Biol. Reprod. 38, 1171–1180), whereas embryos were cultured in sequential media (Lane et al. 2005 Theriogenology 60, 407–419). All embryos that cleaved by 30 hpi (early cleavers, EC) were selected and cultured separately. The remaining embryos cleaved by 48 hpi (non early cleavers, NEC) were also incubated in separate drops. Blastocysts of proper morphology were collected at 176 hpi and subjected to sex determination by PCR (AMGL gene). The significance of developmental stage, timing of the first zygotic cleavage and GH supplementation in relation to the sex ratio was evaluated by the chi-square test. All straws with frozen semen of 2 bulls used in this experiment were derived from single ejaculates. The sex ratio of sperm samples used for IVF was evaluated by FISH with locus-specific probes. The experiment was done on 266 embryos obtained from 1249 oocytes. A significant predominance of male blastocysts regardless of the experimental conditions was observed [the male to female ratio (M:F) 2:1, P < 0.001]. FISH analysis revealed that there was no deviation from the expected 1:1 ratio of X and Y spermatozoa in sperm samples used for IVF. When the timing of the first zygotic cleavage was considered, shift in M:F ratio in favor of males (P < 0.01) in blastocysts derived from EC zygotes was noticed. Due to a very low number of NEC embryos, this category was not included in analysis. Although the M:F ratio was shifted towards males, the rate of female embryos was greater in the control group (25F/38M) v. GH group (16F/39M) of EC expanded blastocysts. This phenomenon was not observed among hatched blastocysts; however, GH presence caused an increase (P < 0.01) in the number of females at this stage. Therefore GH may stimulate embryonic growth, especially embryos of reduced quality, through its positive influence on cytoplasmic oocyte maturation. In conclusion, the predominance of male blastocysts observed in this study may be attributed to the applied IVF procedure because the X:Y ratio in spermatozoa was not different from the expected 1:1 ratio. Moreover, significantly fewer females among analyzed blastocysts may suggest that the developmental potential of female embryos in applied in vitro conditions was somehow reduced when compared with males. This became evident when the transition from expanded to hatched blastocysts was observed. This work was supported by the project No. N302 046 31/3780 of the Ministry of Science and Higher Education, Poland.

Mechanism of the adverse effect of hyaluronidase used for oocyte denudation on early development of bovine embryos

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Shiori Ashibe ◽  
Kanade Irisawa ◽  
Ken Yokawa ◽  
Yoshikazu Nagao
Keyword(s):  
Treatment Group ◽  
Assisted Reproductive Technologies ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Control Group ◽  
Free Calcium ◽  
Parthenogenetic Development ◽  
Early Embryos ◽  
Bovine Oocytes

Summary Hyaluronidase is widely used in animal and human assisted reproductive technologies (ARTs) to remove cumulus cells around oocytes. However, adverse effects of hyaluronidase treatment, such as increased rates of degeneration and parthenogenesis, have been found after treatment of human and mouse oocytes. Currently, the mechanism(s) of the detrimental effects are unclear. The present study was initiated to identify the mechanism of adverse responses to hyaluronidase treatment in bovine oocytes and early embryos. Cumulus cells were removed from cumulus–oocyte complexes (COCs) with or without hyaluronidase and the oocytes were subjected to intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Significantly lower rates of blastocyst formation were obtained in the hyaluronidase treatment group after ICSI (22.4%) and IVF (21.2%) compared with the non-hyaluronidase control groups: 36.1% after ICSI and 30.4% after IVF. Next, we examined the effect of hyaluronidase on parthenogenetic development rates and on the cytoplasmic levels of free calcium ions (Ca2+), reactive oxygen species (ROS) and reduced glutathione (GSH). No differences in parthenogenesis rates were found between treated and untreated groups. Ca2+ levels in oocytes from the hyaluronidase treatment group indicated using mean fluorescence intensity were significantly higher (68.8 ± 5.3) compared with in the control group (45.0 ± 2.5). No differences were found in the levels of ROS or GSH between the treated and untreated groups. We conclude that hyaluronidase might trigger an increase in Ca2+ levels in oocytes, resulting in a decreased potential for normal embryonic development.

Novel aspects of endometrial function: a biological sensor of embryo quality and driver of pregnancy success

2012 ◽  
Vol 24 (1) ◽  
pp. 68 ◽  
Author(s):  
Olivier Sandra ◽  
Nadéra Mansouri-Attia ◽  
Richard G. Lea
Keyword(s):  
Developmental Stages ◽  
Assisted Reproductive Technologies ◽  
Uterine Cavity ◽  
Reproductive Technologies ◽  
Biological Properties ◽  
Reproductive Capacity ◽  
Epigenetic Alterations ◽  
Uterine Receptivity ◽  
Blastocyst Development ◽  
Developmental Potential

Successful pregnancy depends on complex biological processes that are regulated temporally and spatially throughout gestation. The molecular basis of these processes have been examined in relation to gamete quality, early blastocyst development and placental function, and data have been generated showing perturbations of these developmental stages by environmental insults or embryo biotechnologies. The developmental period falling between the entry of the blastocyst into the uterine cavity to implantation has also been examined in terms of the biological function of the endometrium. Indeed several mechanisms underlying uterine receptivity, controlled by maternal factors, and the maternal recognition of pregnancy, requiring conceptus-produced signals, have been clarified. Nevertheless, recent data based on experimental perturbations have unveiled unexpected biological properties of the endometrium (sensor/driver) that make this tissue a dynamic and reactive entity. Persistent or transient modifications in organisation and functionality of the endometrium can dramatically affect pre-implantation embryo trajectory through epigenetic alterations with lasting consequences on later stages of pregnancy, including placentation, fetal development, pregnancy outcome and post-natal health. Developing diagnostic and prognostic tools based on endometrial factors may enable the assessment of maternal reproductive capacity and/or the developmental potential of the embryo, particularly when assisted reproductive technologies are applied.

Effectiveness of an Educational Intervention to Participation in Physical Activity: A quasi-experimental model-based study

2020 ◽  
Author(s):  
Saeideh Shahsavari ◽  
Sakineh dadipoor ◽  
Mohtasham Ghaffari ◽  
Ali Safari-Moradabadi
Keyword(s):  
Physical Activity ◽  
Educational Intervention ◽  
Stages Of Change ◽  
Intervention Group ◽  
T Test ◽  
Control Group ◽  
Post Intervention ◽  
Chi Square ◽  
Significant Difference ◽  
Quasi Experimental

Abstract Background: The aim of the present study was to assess readiness to become or stay physically active according to the Stages of Change Model.Methods: The present quasi-experimental study was conducted on 100 women working in the healthcare centres of Bandar Abbas, Iran. The sampling method is clustering in type. The subjects were assigned into two groups of intervention and control. The collected data were analysed by SPSS-16 software using descriptive and inferential statistics, including independent-sample t-test, paired-sample t-test and Chi-square test.Results: Before the educational intervention, 19 subjects (0.38%) from the intervention group showed to have regular physical activity (4-5 stages). This number changed to 29 (0.58%) and 25 (0.50%) after three months and six months of intervention. A statistically significant difference was found before the intervention and 3 and 6 months afterwards (P˂.001). In the control group, no statistically significant difference was found between the pre-intervention and post-intervention (three months (P=.351) and six months (P=.687).Conclusion: The educational intervention based on the stages of behaviour change model showed to be effective in promoting the physical activity of employed women. These findings may benefit health education researchers and practitioners who tend to develop innovative theory-based interventions and strategies to increase the level of physical activity in women.

71 EFFECT OF CELL QUANTITY IN MICE CHIMERA PRODUCTION BY DEMI-BLASTOCYST AGGREGATION

2012 ◽  
Vol 24 (1) ◽  
pp. 148
Author(s):  
C. Pontes Godoi ◽  
P. D. Moço ◽  
B. Cazari ◽  
P. T. Mihara ◽  
P. V. Silva ◽  
...  
Keyword(s):  
Cell Mass ◽  
Farm Animals ◽  
Control Group ◽  
Chi Square ◽  
Cell Stage ◽  
Exact Test ◽  
Chi Square Test ◽  
Aggregation Technique

Eight-cell-stage to pre-compaction morula are the most used embryonic stages to aggregation, because the embryos, in these early stages, synthesise cell adhesion molecules that increase the aggregation chances among them (Vestweber et al. 1987 Develop. Biol. 124, 451–456). Although post-compaction embryos produce reduced aggregation rates, they are not refractory to this process (Nogueira et al. 2010 Transgenic Res. 19, 344–345). Based on the evidence of less permissive aggregation in post-compaction-stage embryos and the need to expose the inner surface of those embryos to improve aggregation rate, the aim of this study was to evaluate, in mice, the influence of cell quantity (i.e. the quantity of half-embryos put together to aggregate themselves) in the chimerism rate of split blastocysts. Embryos, with preferentially different phenotypes, were obtained from C57BL/6/EGFP and Swiss Webster strains. Females ranging from 21 to 45 days old were superstimulated and mated according to Mancini et al. (2008 Transgenic Res. 17, 1015). Eight-cell-stage embryos (8C) and pre-compaction morula (PCM) were recovered (2 to 2.5 days post coitum) and had their zona pellucida removed using pronase treatment (2 mg mL–1 for 15 min), whereas blastocysts (recovered 3.5 dpc) were split with a microblade controlled by micromanipulator in an inverted microscope (NK2; Eppendorf, Hamburg, Germany and Eclipse Ti; Nikon, Tokyo, Japan, respectively). The aggregation groups were a control (C) with 2 pre-compaction whole embryos (8C or PCM, or both) and 2 experimental with post-compaction embryos [i.e. 2 (2DB) or 4 (4DB) demi-blastocysts]. The structures (2 or 4) of the groups were stuck to each other with the use of phytohemagglutinin (1 mg mL–1) and cultured in vitro by 24 h (37°C, 5% CO2 and saturated humidity). After culture, the presence of chimeric embryos was verified by detection of a single, cohesive cell mass or a structure in an 8 shape with more than one-half of its total diameter aggregated. For the 4DB group, a successful aggregation was considered when, at least 2 of 4 DB had aggregated. The results were analysed using chi-square test, Fisher's exact test and Kruskal-Wallis (to compare among groups, between groups and among medians of group replicates, respectively) and significance was considered when P < 0.05. The aggregation rates for the groups C, 2DB and 4DB were, respectively, 77.3a; 8.3b and 36.4%c (P < 0.001). The increasing of the aggregation technique efficacy, in post-compaction stages, would be particularly interesting in farm animals (e.g. bovine species), where it is not feasible to obtain, in vivo, pre-compaction stages embryos (as 8 cells) and when only trophectoderm aggregation is wanted. It was concluded that cell increasing (from 2 to 4 DB) improved the chimerism rate, but not enough to be similar to the control group. Supported by FAPESP of Brazil.

scholarly journals Gender Differences in the Experience of Infertility Concerning Polish Couples: Preliminary Research

2019 ◽  
Vol 16 (13) ◽  
pp. 2337 ◽  
Author(s):  
Małgorzata Nagórska ◽  
Anna Bartosiewicz ◽  
Bogdan Obrzut ◽  
Dorota Darmochwał-Kolarz
Keyword(s):  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Clinical Pregnancy ◽  
World Health ◽  
Independence Test ◽  
Chi Square ◽  
Unprotected Sexual Intercourse ◽  
The World ◽  
Preliminary Research ◽  
Health Organization

The World Health Organization (WHO) determines infertility as a disease of the reproductive system defined clinically by the failure to achieve a clinical pregnancy after 12 months or more of regular unprotected sexual intercourse. Estimates indicate that the problem of infertility in the world is continuing to grow. The aim of the study was to compare approaches to disease in partners of both sexes diagnosed with infertility. The study was conducted among 61 couples treated for infertility using an original questionnaire developed by the authors. The Chi square independence test was used for statistical analysis. Both men and women responded to the diagnosis of infertility with negative emotions. Regardless of sex, sadness and anxiety were the dominant feelings associated with the diagnosis of infertility. Women believed in the success of the treatment to a greater extent than men. Mainly women attempted to talk openly about the problem of infertility, while men were more restrained in this respect. Women accepted the assisted reproductive technologies (ART) to a greater extent than men, but men would accept childlessness more often than women.

scholarly journals The Effect of Using the Care Model of "Sensitization" on Medication Adherence in Asthmatic Patients

2016 ◽  
Vol 8 (11) ◽  
pp. 11 ◽  
Author(s):  
Mahbobeh Sajadi ◽  
Mozhgan Bagheri ◽  
Davood Hekmatpou ◽  
Hamid Borsi
Keyword(s):  
Medication Adherence ◽  
T Test ◽  
Control Group ◽  
Care Model ◽  
Chi Square ◽  
Asthmatic Patients ◽  
Significant Difference ◽  
Prevalent Disease ◽  
And Control ◽  
Positive Effect

<p><strong>INTRODUCTION:</strong> Asthma is a prevalent disease with a multitude of complication. The present research was aimed at investigating the effects of implementing sensitization care model on medication adherence in asthmatic patients.</p><p><strong>METHODS &amp; MATERIALS:</strong> In this interventional study, 74 subjects were selected using accessibility sampling method and were randomly classified into 2 separate groups, intervention (37 subjects) and control group (37 subjects). Data were collected with questionnaire. A month later, the subjects in both groups (control and intervention) completed questionnaire again. The collected data were analyzed using SPSS software, Independent t-test, Paired t-test and Chi-Square.</p><p><strong>RESULTS:</strong> Data analysis showed that the average age of participants was 41 years (range 21-83). The most and lowest frequencies of marital status in both sexes were 66.2% married and 33.8% single, respectively. There was no significant difference before intervention between two groups. However, after the intervention, there was a significant difference between the two groups (P=0.0001).</p><p><strong>DISCUSSION:</strong> The results of the present study showed that implementing sensitization care model had a positive effect on medication adherence among asthmatic patients. Therefore, application of this model is recommended to care and treatment asthmatic patients.</p>

Effectiveness and outcomes of embryo cryopreservation programs in assisted reproductive technologies

2014 ◽  
Vol 63 (4) ◽  
pp. 39-46 ◽  
Author(s):  
Yana Nikolayevna Kravchuk ◽  
Alla Stanislavovna Kalugina ◽  
Olga Vladimirovna Bystrova ◽  
Svetlana Aleksandrovna Shlykova
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Negative Impact ◽  
Perinatal Outcomes ◽  
Reproductive Technologies ◽  
Embryo Cryopreservation ◽  
Control Group ◽  
Fresh Embryo Transfer ◽  
Art Programs ◽  
Thawed Embryo Transfer

Background. Embryo cryopreservation is an essential part of ART programs today. In recent years vitrification method is used increasingly widely. Purposes and tasks. To compare the effectiveness of ART programs using vitrified and fresh embryos, as well as different endometrial preparation regimes for frozen\thawed embryo transfer (modified natural cycle (MNC) and the preparatory hormone therapy(PHT)). To analyze the course of pregnancy and perinatal outcomes after vitrified embryo transfer. Materials and methods. We prospectively assessed the ART programs effectiveness and perinatal outcomes in 153 patients (I group), who underwent vitrified embryo transfer in 2011-2013 year. To prepare the endometrium for thawed embryo transfer in 83 patients PHT (Ia subgroup) and MNC in 70 patients (Ib subgroup) were used. Control group consisted of 70 patients, who underwent fresh embryo transfer. Results. The clinical pregnancy rate, birth rate and “take home baby” rate were not significantly different between the I (47,5 %; 30,9 %; 30,9 %) and II (53,0 %; 34,9 %; 32,5 %) groups, and between Ia (48,3 %; 28,4 %; 28,4 %) and IIb (46,6 %; 34,1 %; 34,1 %) subgroups. Complications during pregnancy and delivery, birthweight, length, Apgar score, congenital malformation rate did not differ significantly after vitrified and fresh embryo transfer. Conclusion. Vitrification is an effective method to achieve clinical results, comparable to native cycles. Application of PHT and MNC results in similar clinical outcomes. Transfer Vitrified embryo transfer does not have a negative impact on obstetric and perinatal outcomes when compared with native cycles.

scholarly journals Pisang Gen Alfa: A Smartphone Application To Reduce Parenting Stress For Parents With Alpha Generations

2020 ◽  
Vol 3 (3) ◽  
pp. 121
Author(s):  
Ahmad Guntur Alfianto ◽  
Nicky Danur Jayanti
Keyword(s):  
Parenting Stress ◽  
Parental Stress ◽  
Sampling Technique ◽  
Smartphone Application ◽  
T Test ◽  
Control Group ◽  
Average Difference ◽  
Chi Square ◽  
Control Group Design ◽  
Quasi Experimental

Parenting stress experienced by parents for alpha generation. Efforts that can be done are to reduce parenting stress with the right techniques. One technique to reduce stress is PISANG GEN ALFA/Mobile Application Parenting Assertive and Management Stress. The purpose of this study was to analyse the effect of PISANG GEN ALFA on parenting stress for parents with alpha generation. The method used was a quasi-experimental pre-test-post-test with control group design with a sample of 100 parents with alpha generation. The sampling technique was purposive sampling with research instruments using the Parental Stress Scale (PSS). The research was conducted in Malang City and Batu City. The statistical test used Chi-Square Tes, Paired Sample T-Test and independent T-test. The results of the distribution of characteristics were not different between the two groups before and after being given treatment. There was an effect of reducing parenting stress after being given PISANG GEN ALFA (0,000) and there was an average difference between the treatment and control groups (0.001). PISANG GEN ALFA is one of the mental health applications that can reduce parenting stress in parents with alpha generation. So that there was a reduction in parenting stress after being given PISANG GEN ALFA and there was an average difference between the group using PISANG GEN ALFA compared to the control group in reducing parenting stress for parents with generations of the children.

scholarly journals Perubahan Respon Pendengaran Karena Pemakaian Earphone

2019 ◽  
Vol 21 (1) ◽  
pp. 20
Author(s):  
Dewi Rossalia
Keyword(s):  
Call Center ◽  
T Test ◽  
Control Group ◽  
Chi Square ◽  
Control Group Design ◽  
Anova Test ◽  
Whitney Test ◽  
Group Design ◽  
Post Test ◽  
Chi Square Test

Abstrak Telah dilakukan penelitian tentang perubahan respon pendengaran karena pemakaian earphone terhadap karyawan bagian call center yang meliputi pengukuran intensitas kebisingan pada earphone yang biasa digunakan oleh karyawan, pengisian kuesioner dan pengukuran ambang pendengaran dengan audiometer. Penelitian ini bertujuan untuk  mengetahui pengaruh kebisingan suara yang dikeluarkan oleh earphone terhadap ambang dengar manusia, mengetahui adanya hubungan lama masa kerja dan usia terhadap gangguan pendengaran pada karyawan call center. Rancangan penelitian eksperimen dengan Randomized Post Test Only Control Group Design. Setelah dilakukan pengambilan data intensitas kebisingan yang dikeluarkan oleh earphone adalah sekitar 59-62 dB. Untuk mengkaji pengaruh intensitas kebisingan terhadap pendengaran karyawan dilakukan pengukuran audiometer pada karyawan yang terpapar kebisingan earphone (kelompok perlakuan) dan karyawan yang tidak terpapar kebisingan earphone (kelompok kontrol). Data-data yang dihasilkan diolah dengan menggunakan uji T dua sampel bebas (chi-square t-test), Chi-Square test, Anova test dan Mann-Whitney test. Dari hasil penelitian didapatkan hubungan yang bermakna antara kebisingan earphone terhadap ambang pendengaran karyawan kelompok perlakuan dengan kelompok kontrol (p<0,05). Kata kunci : kebisingan earphone, gangguan pendengaran.

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