• Title/Summary/Keyword: fertilization methods

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Development of Electronic Mapping System for N-fertilizer Dosage Using Real-time Soil Organic Matter Sensor (실시간 토양 유기물 센서와 DGPS를 이용한 질소 시비량 지도 작성 시스템 개발)

  • 조성인;최상현;김유용
    • Journal of Biosystems Engineering
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    • v.27 no.3
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    • pp.259-266
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    • 2002
  • It is crucial to know spatial soil variability for precision farming. However, it is time-consuming, and difficult to measure spatial soil properties. Therefore, there are needs fur sensing technology to estimate spatial soil variability, and for electronic mapping technology to store, manipulate and process the sampled data. This research was conducted to develop a real-time soil organic matter sensor and an electronic mapping system. A soil organic matter sensor was developed with a spectrophotometer in the 900∼1,700 nm range. It was designed in a penetrator type to measure reflectance of soil at 15cm depth. The signal was calibrated with organic matter content (OMC) of the soil which was sampled in the field. The OMC was measured by the Walkeley-Black method. The soil OMCs were ranged from 0.07 to 7.96%. Statistical partial least square and principle component regression analyses were used as calibration methods. Coefficient of determination, standard error prediction and bias were 0.85 0.72 and -0.13, respectively. The electronic mapping system was consisted of the soil OMC sensor, a DGPS, a database and a makeshift vehicle. An algorithm was developed to acquire data on sampling position and its OMC and to store the data in the database. Fifty samples in fields were taken to make an N-fertilizer dosage map. Mean absolute error of these data was 0.59. The Kring method was used to interpolate data between sampling nodes. The interpolated data was used to make a soil OMC map. Also an N-fertilizer dosage map was drawn using the soil OMC map. The N-fertilizer dosage was determined by the fertilizing equation recommended by National Institute of Agricultural Science and Technology in Korea. Use of the N-fertilizer dosage map would increase precision fertilization up to 91% compared with conventional fertilization. Therefore, the developed electronic mapping system was feasible to not only precision determination of N-fertilizer dosage, but also reduction of environmental pollution.

Application of two different synthetic sequential media for the human IVF-ET program: a prospective, randomized, and comparative study

  • Yoon, Jeong;Yoon, Hye-Jin;Juhn, Kyoung-Mi;Ko, Jin-Kyung;Yoon, San-Hyun;Ko, Yong;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.186-192
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    • 2011
  • Objective: Since IVF program was first established, various types of media and culture systems have been developed either in-house or commercially. The aim of this study was to compare the efficacy of in-house Maria Research Center (MRC) media to that of commercially available Sydney IVF media in human day 3 embryo transfer cycles. Methods: Three hundred sixty nine couples were included in this prospective, randomized, and comparative study. All couples undergoing IVF treatment at the Maria Fertility Hospital were randomly assigned to either Sydney IVF (n=178) or MRC (n=191) media. Results: No difference was observed between the MRC media and Sydney IVF media groups with respect to fertilization rate (74.4% vs. 75.5%). The clinical pregnancy and implantation rates of MRC media (47.1% and 20.0%, respectively) were also similar to those of Sydney IVF media (44.4% and 19.4%, respectively). However, the proportion of embryos with good quality on day 3 was significantly higher in the MRC media group than the Sydney IVF media group (50.2% vs. 43.2%) ($p$ <0.05). Conclusion: MRC media were as effective as Sydney IVF media for sustaining embryo development and pregnancy rates. The present study implies that MRC media can be a suitable alternative to commercially available media for human IVF-ET program.

Effects of dynamic oxygen concentrations on the development of mouse pre- and peri-implantation embryos using a double-channel gas supply incubator system

  • Lee, Seung-Chan;Seo, Ho-Chul;Lee, Jaewang;Jun, Jin Hyun;Choi, Kyoo Wan
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.4
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    • pp.189-196
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    • 2019
  • Objective: We aimed to evaluate the effects of different oxygen conditions (20% [high O2], 5% [low O2] and 5% decreased to 2% [dynamic O2]) on mouse pre- and peri-implantation development using a novel double-channel gas supply (DCGS) incubator (CNC Biotech Inc.) to alter the oxygen concentration during in vitro culture. Methods: The high-O2 and low-O2 groups were cultured from the one-cell to the blastocyst stage under 20% and 5% oxygen concentrations, respectively. In the dynamic-O2 group, mouse embryos were cultured from the one-cell to the morula stage under 5% O2 for 3 days, followed by culture under 2% O2 to the blastocyst stage. To evaluate peri-implantation development, the blastocysts from the three groups were individually transferred to a fibronectin-coated dish and cultured to the outgrowth stage in droplets. Results: The blastocyst formation rate was significantly higher in the low-O2 and dynamic-O2 groups than in the high-O2 group. The total cell number was significantly higher in the dynamic-O2 group than in the low-O2 and high-O2 groups. Additionally, the apoptotic index was significantly lower in the low-O2 and dynamic-O2 groups than in the high-O2 group. The trophoblast outgrowth rate and spread area were significantly higher in the low-O2 and dynamic-O2 groups than in the high-O2 group. Conclusion: Our results showed that a dynamic oxygen concentration (decreasing from 5% to 2%) had beneficial effects on mouse pre- and peri-implantation development. Optimized, dynamic changing of oxygen concentrations using the novel DCGS incubator could improve the developmental competence of in vitro cultured embryos in a human in vitro fertilization and embryo transfer program.

The prevalence of positive urinary cotinine tests in Korean infertile couples and the effect of smoking on assisted conception outcomes

  • Kim, Hoon;Kim, Seul Ki;Yu, Eun Jeong;Lee, Jung Ryeol;Jee, Byung Chul;Suh, Chang Suk;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.4
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    • pp.136-142
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    • 2015
  • Objective: Smoking has been reported to harm nearly every organ of the body, but conflicting results have been reported regarding the effects of smoking on assisted conception. In this prospective cohort study, we aimed to investigate the prevalence of positive urinary cotinine tests in infertile couples and whether cotinine positivity was associated with infertility treatment outcomes. Methods: A qualitative urinary cotinine test was administered to 127 couples who underwent in vitro fertilization (IVF, n=92) or intrauterine insemination (IUI, n=35). Results: The overall prevalence of positive urinary cotinine test was 43.3% (55/127) in the male partners and 10.2% (13/127) in the female partners with similar prevalence rates in both genders in the IUI and IVF groups. Semen characteristics, serum markers of ovarian reserve, and number of retrieved oocytes were comparable among cotinine-positive and cotinine-negative men or women (with the exception of sperm count, which was higher among cotinine-positive men). The results of urinary cotinine tests in infertile couples were not associated with IVF and IUI outcomes. Conclusion: The presence of cotinine in the system, as indicated by a positive urinary cotinine test, was not associated with poorer outcomes of infertility treatment.

Optimal numbers of mature oocytes to produce at least one or multiple top-quality day-3 embryos in normal responders

  • Shim, Yoo Jin;Hong, Yeon Hee;Kim, Seul Ki;Jee, Byung Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.3
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    • pp.221-226
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    • 2020
  • Objective: We attempted to identify the optimal cutoff numbers of mature oocytes that would produce at least one or multiple top-quality (grade A) day-3 embryos in normal responders undergoing stimulated in vitro fertilization (IVF) cycles. Methods: We selected 210 fresh IVF cycles performed in 170 infertile women at a single center from January 2014 to November 2019. Four to 14 (total) oocytes were obtained in all cycles after conventional ovarian stimulation. A receiver operating characteristic curve analysis was performed to find the moderate and extreme cutoff numbers of mature oocytes that would produce ≥ 1, ≥ 2, ≥ 3, ≥ 4, and ≥ 5 top-quality embryos. Results: The cutoff number of mature oocytes was significantly correlated with the number of top-quality embryos (r = 0.467, p= 0.000). The moderate cutoff number of mature oocytes was ≥ 3, ≥ 5, ≥ 5, ≥ 6, and ≥ 6 for obtaining ≥ 1, ≥ 2, ≥ 3, ≥ 4, and ≥ 5 top-quality embryos, respectively. The extreme cutoff number of mature oocytes was ≥ 9, ≥ 9, ≥ 10, ≥ 10, and ≥ 11 for obtaining ≥ 1, ≥ 2, ≥ 3, ≥ 4, and ≥ 5 top-quality embryos, respectively. Conclusion: We present the optimal cutoff numbers of mature oocytes that would yield ≥ 1, ≥ 2, ≥ 3, ≥ 4, and ≥ 5 top-quality embryos with 95% specificity. Our findings could help infertility clinicians to set target mature oocyte numbers in women undergoing stimulated IVF cycles.

Comparison of Two Vitrification Methods of In Vitro Development Oocytes Collected from Porcine Antral Follicles Using Open Pulled Straw (OPS) Techniques

  • An, Mihyun;Hong, Daewuk;Son, Dongsoo;Seok, Hobong
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.84-84
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    • 2002
  • The advantages of the OPS techniques(Vajta G et al, Mol Reprod Dev 51: 53-58,1998) give 1) high survival rates of various types of eggs, 2) quick and simple process, 3) inexpensive equipment and reduced chilling injury. The efficiency of IVM/IVF technique in the porcine species is relatively lower than that obtained in other species such as ruminants. Two experiments were designed to investigate the effects of in-vitro fertilization of porcine oocytes matures using different OPS protocol for chilling and warming of vitrification. Porcine oocytes from ovaries collected at abattoir were matured for 44 hours in TCM199 Earle's salt supplemental with pyruvate, pff, L-cysteine, hormones and gentamycin. Oocytes were denuded and fertilized with frozen boar semen by common method. Porcine embryos produced routinely by in-vitro culture system of NCSU23 medium. The vitrification and the warming were conducted by OPS method with the glass micropipette instead of straw vessels and modified the protocol of G.Vajta(1999). In Exp 1, Chilling/Warming:Holding Medium(HM)+EG+DMSO/HM +sucrose Medium(SM) at 39$^{\circ}C$ warm stage. In Exp 2, : PBS+CS+EG+Ficoll+ Trehalose/PBS+Trehalose at 25$^{\circ}C$ stage. Filling, freezing, packing, thawing out and further culturing were performed to follow the basic protocol of G Vajta. During IVM-lVC and post-warming, fertilization parameter and developmental potential were compared to and statistically analysed. It was not significantly different from Exp 1 and Exp 2 but 25$^{\circ}C$ of stage was slightly higher on the morula/blastocyst forming rate and better atmosphere for worker than that at 39$^{\circ}C$ stage.

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Improving quality of common reed (Phragmites communis Trin.) silage with additives

  • Asano, Keigo;Ishikawa, Takahiro;Araie, Ayako;Ishida, Motohiko
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.11
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    • pp.1747-1755
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    • 2018
  • Objective: Common reed (Phragmites communis Trin.) could potentially provide an alternative resource for silage; however, its silage quality is poor. The aim of this study was to investigate the factors in reed that contribute to poor quality and determine how the use of additives at ensiling could improve fermentation quality. Methods: In Experiment 1, we determined the chemical composition and the presence of indigenous lactic acid bacteria (LAB) in reed. We further examined fermentation quality of reed silage under conditions without additives (NA) and treated glucose (G), lactic acid bacteria (L), and their combination (G+L). In Experiment 2, silage of NA, and with an addition of cellulase and lactic acid bacteria (CL) were prepared from harvested reed. The harvested reeds were fertilized at nitrogen concentrations of 0, 4, 8, and $12g\;N/m^2$ and were harvested thrice within one year. Results: The indigenous LAB and fermentable carbohydrates are at extremely low concentrations in reed. Reed silage, to which we added G+L, provided the highest quality silage among treatments in Experiment 1. In Experiment 2, N fertilization had no negative effect on silage quality of reed. The harvest times decreased fermentable carbohydrate content in reed. The CL treatment provided a higher lactic acid content compared to the NA treatment. However, the quality of CL treated silage at the second and third harvests was significantly lower than at the first harvest, due to a reduction in carbohydrates caused by frequent harvesting. Conclusion: The causes of poor quality in reed silage are its lack of indigenous LAB and fermentable carbohydrates and its high moisture content. In addition, reed managed by frequent harvesting reduces carbohydrate content. Although the silage quality could be improved by adding CL, higher-quality silage could be prepared by adding fermentable carbohydrates, such as glucose (rather than adding cellulases).

Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

  • Park, Kyunghyuk;Frost, Jennifer M.;Adair, Adam James;Kim, Dong Min;Yun, Hyein;Brooks, Janie S.;Fischer, Robert L.;Choi, Yeonhee
    • Molecules and Cells
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    • v.39 no.10
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    • pp.768-775
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    • 2016
  • The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

Functional role of Forskolin and PD166285 in the development of denuded mouse oocytes

  • Cao, Hongguo;Bian, Yani;Zhang, Fei;Tang, Yunshu;Li, Caixia;Chen, Jiemei;Zhang, Xiaorong
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.344-353
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    • 2018
  • Objective: cAMP and mature promoting factor (MPF) play critical roles during the maturation of mammalian oocytes. The aim of this study was to produce the offspring from denuded oocytes (DOs) in mice by regulating cAMP and MPF. Methods: In this study, we used DOs at the germinal vesicle (GV) stage in mice and regulated levels of cAMP and MPF in DOs by adding Forskolin and PD166285 during in vitro maturation without follicle stimulating hormone and luteinizing hormone, respectively. Results: Combined use of $50{\mu}M$ Forskolin for 3 h and $2.5{\mu}M$ PD166285 for additional 21 h enhanced the developmental competence of DOs, maturation rate of DOs was $76.71%{\pm}4.11%$, blastocyst rate was $18.33%{\pm}4.44%$ after parthenogenetic activation (PA). The DOs could successfully be fertilized with sperm in vitro, cleavage rate was $17.02%{\pm}5.82%$ and blastocyst rate was $5.65%{\pm}3.10%$. Besides, 2-cell in vitro fertilization embryos from DOs produced 4 normal live offspring (4/34). Conclusion: The results confirmed that the combination of Forskolin and PD166285 can induce DOs to complete meiosis process and produce normal offspring.

Cryopreservation of Oocytes and Embryos by Vitrification (유리화 방법에 의한 난자와 수정란의 동결보존)

  • Gupta, Mukesh Kumar;Lee, Hoon-Taek
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.267-291
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    • 2010
  • Life can be kept in suspended animation either before fertilization at oocyte stage or after fertilization at different stages of embryonic development for a variety of reasons. It not only has potential applications in fertility preservation and management in human but also has important roles in the preservation and management of animal genetic resources, low-cost international movement of selected genetics, and rapid dissemination of germplasm through assisted reproductive technologies (ART) and genetic engineering. Currently, slow-freezing and vitrification are the two approaches by which oocytes and embryos can be cryopreserved for long-term storage. Both of these methods have their own advantages and disadvantages but allow the cryopreservation of oocytes and embryos with comparable efficiency. Vitrification of oocyte and embryos, although proven successful just 13 years after slow-freezing, is generally considered an emerging technology and appears to slow gain acceptance in both animal and human ART despite having controversial storage and contamination issues. In this manuscript, we discuss the basic techniques of oocyt/embryo cryopreservation and review the current status and recent developments in vitrification.