• Title/Summary/Keyword: metaphase

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Analysis of FISH patterns using 5S and 45S rDNAs in Codonopsis minima and C. lanceolata from Jeju Island (5S와 45S rDNA 유전자를 이용한 제주도산 애기더덕 (Codonopsis minima)과 더덕 (C. lanceolata)의 FISH 패턴 분석)

  • Kim, Soo-Young;Kim, Chan-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.186-190
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    • 2010
  • The chromosome number was identified and fluorescence in situ hybridization(FISH) mapping of 5S and 45S rDNAs were conducted for C. minima and C. lanceolata in the genus Codonopsis from Jeju island. In this study, we have confirmed that the somatic metaphase chromosome number determined as 2n=2x=16 was the same as the findings from the previous studies. While the conventional staining method makes it rather difficult to distinguish satellite chromosomes due to high degree of variability, FISH analysis produced the exact number and location of 5S and 45S rDNAs. Both species in the genus Codonopsis have a pair of 5S rDNA and their gene loci were observed on chromosome 3. Although two pairs of 45S rDNAs (one on chromosome 1 and the other on chromosome 8) were identified in both species, the 45S rDNA signals on chromosome 8 in C. minima were significantly weaker than those on chromosome 1. In addition, the 45S rDNA signals on chromosome 1 in C. lanceolata showed that the chromosome is non-homologus. In this study, we have determined cytogenetic characteristics of C. minima and C. lanceolata according to their gene replication patterns.

Preferred strategy for euploid single embryo transfer in advanced maternal age: Fresh versus frozen

  • Fatma Ozdemir;Gokalp Oner;Semra Kahraman;Yucel Sahin;Hakan Yelke
    • Clinical and Experimental Reproductive Medicine
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    • v.51 no.1
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    • pp.85-90
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    • 2024
  • Objective: The purpose of this study was to compare fresh and frozen-thawed euploid blastocyst transfer protocols following preimplantation genetic screening (PGS) in cases of advanced maternal age. Methods: A total of 330 patients were examined retrospectively. PGS was performed on the embryos of 146 patients for whom fresh transfers were chosen. In contrast, frozen-thawed euploid single embryo transfer (ET) was selected after PGS for 184 patients, and their embryos were vitrified. The percentage of euploid embryos and rates of implantation, pregnancy, and pregnancy continuity, as well as clinical and biochemical abortion rates, were compared. Results: The numbers of retrieved oocytes, metaphase II oocytes, and fertilized ova were greater in the frozen-thawed group. The percentages of euploid embryos were comparable between the fresh and frozen-thawed groups (32% vs. 34.8%, respectively). The rates of implantation (46.6%vs. 62.5%), pregnancy (50% vs. 66.8%), ongoing pregnancy (38.4% vs. 53.8%), and live birth percentage (37.0% vs. 53.8%) were significantly higher in the frozen-thawed group. However, no significant differences were found in the clinical and biochemical abortion rates. Conclusion: The use of frozen-thawed single euploid ET is associated with increased implantation and pregnancy rates compared to fresh single euploid ET with PGS.

The Karyotype of Payamphistomum explanatum(Creplin, 1849) Obtained from Korean Cattle (한국산 Paramphistomum explanatum(Creplin, 1849)의 핵형분석)

  • Lee, Jae-Gu;Gang, Chang-Won;Lee, Ho-Il
    • Parasites, Hosts and Diseases
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    • v.24 no.1
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    • pp.42-48
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    • 1986
  • As a series of systematic classification of paramphistomes, in the first step, paramphistomes in the lumen and reticulum were collected on 170 Korean catties (2∼3 years age, male) slaughtered at Jeonju abattoir from July, 1984 to September, 1985 and were classified by means of morphology of the worms. Afterwards, the karyotype of Paramphistomum explanatum (Creplin, 1849) which is the common in Korean cattle was detected by means of modified air-drying method from testis cells of the worm. The following is a brief summary of the leading facts gained through the experiment. 1. Most of the cattle slaughtered at the abattoir were infected with paramphistomes. The 5 species of the worms were detected on 170 Korean cattle and the worm burden per head was from 2 to 784(on the average 170) worms, 120(70.59%) heads out of them involving 2∼100 worms. In 28,900 individuals of paramphistomes obtained on 170 Korean cattle, appearance rates of various worms were as follows : 49.74% in P. explanatum, 48.08% in P. cervi, 0.98% in Orthocoelium orthocoelium, 0.89% in Fischoederius cobboldi and 0.14% in Cotylophoron cotylcphorum. 2. The chromosome number of 620 P. explanatum in the haploid and diploid cells was n=9 and 2n=18, and abundant cells in meiotic division were observed; 1,420 haploid and 38 diploid cells were reliable. Nine pairs of mitotic chromosomes were homologous and the chromosomes were composed of aye medium-sized metacentrics(m), subtelocentrics(st) or submetacentrics(sm) and four smallsized subtelocentrics(st) or submetacentrics(am), while meiotic metaphase chromosomes were composed of five medium and four small-sized. 3. The haploid of the testis cells showed C-band in the centromeric region from 8 of them, whereas the remaining chromosome No. 5 included heterochromatin on the tip region, and chromosomes No. 3 and No. 7 showed a remarkable C-band distinguished from other chromosomes.

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Developmental Competence of Intrafollicular Oocytes Derived from Preantral Follicle Culture with Different Protocols after Parthenogenetic Activation

  • Choi, Jung Kyu;Lee, Jae Hee;Lee, Seung Tae;Choi, Mun Hwan;Gong, Seung Pyo;Lee, Eun Ju;Lim, Jeong Mook
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.8
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    • pp.1190-1195
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    • 2007
  • This study was conducted to improve efficiency of a follicle culture system without reducing developmental competence of intrafollicular oocytes. Preantral follicles (100 to $125{\mu}m$ in diameter) of F1 hybrid (B6CBAF1) mice were cultured singly for 216 h in modified ${\alpha}$-MEM-glutamax medium, to which 2.5 IU/ml hCG and epidermal growth factor was added 16 h prior to the end of culture. Medium change was either performed three times (54 h interval), twice (72 h interval), once (108 h interval), or not at all (216 h interval). Maturation (progression to the metaphase II stage) of intrafollicular oocytes was detected from 4 days after culture in the three-times change treatment, while all treatments yielded mature oocytes from day 5 of culture. Compared with the three-times change, decreasing the change frequency to once did not reduce the capacity to begin maturation (germinal vesicle breakdown of 82 to 86%), to mature (78 to 79%) and to develop into blastocysts after parthenogenetic activation (29 to 32%). Morphological parameters were similar among these treatments. Except for the no medium change treatment, similar colony-forming activity of inner cell mass cells after culturing of blastocysts in leukemia inhibitory factor-containing medium was detected, while the morphology of the colony-forming cells deteriorated in the change-once treatment compared with the change twice or three-times. In conclusion, the efficiency of the preantral follicle culture system could be improved by reducing frequency of medium change up to a 72 h interval (three times in total 216 h culture) without decreasing developmental competence of oocytes.

Studies on a Tetraploid of Robinia pseudoacacia L. Selected in the Planted Forest (조림지(造林地)에서 선발(選拔)한 자연(自然) 4배체(倍體) 아까시나무(Robinia pseudoacacia L.)에 관(關)한 연구(硏究))

  • Kim, Chung Suk;Lee, Suk Koo
    • Journal of Korean Society of Forest Science
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    • v.14 no.1
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    • pp.39-44
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    • 1972
  • Cytological investigation was done on a tetraploid tree of Robinia pseudoacacia L. selected in the planted forest in Chilbo Mountain. From the results of observation of chromosomes on mitotic metaphase in leaves, it was confirmed that the mutant is tetraploid plant (2n=40). The external form of leaves of the spontaneous tetraploid was gigantic and dark green one in comparison with diploid. There was no difference in the size of guard cell and the number of stomata per unit area between the mutant and the diploid plant, however, length and thickness of wood fiber were increased as compared with that of diploid. The spontaneous gigas tetraploid showed growth performance as much as the colchicine induced gigas tetraploid.

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Genotoxicity and Interference with Cell Cycle Activities by an Ethanolic Extract from Thai Plumbago indica Roots in Human Lymphocytes in vitro

  • Thitiorul, Sumon;Ratanavalachai, Treetip;Tanuchit, Sermkiat;Itharat, Arunporn;Sakpakdeejaroen, Intouch
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2487-2490
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    • 2013
  • In Thai traditional medicine, Plumbago indica or Jetamul-Pleung-Dang in Thai is known to have health benefit especially for anti-inflammatory, antibacterial, and antitumor activities. However, the mechanisms of its action are still uncertain. One of which might be genotoxic effects. In the present study, we investigated the genotoxicity of an ethanolic extract of Plumbago indica root (EEPIR) by sister chromatid exchange (SCE) assay in human lymphocytes. Results have shown that all treatments with EEPIR ($12.5-100{\mu}g/ml$) could induce cell cycle delay as shown by significant increase in the number of metaphase cells in the first cell cycle but neither in the second nor the third cell cycle. Only at concentrations of 25, 50, and $100{\mu}g/ml$ were SCE levels significantly increased above that of the control (p<0.05). EEPIR at a concentration of $500{\mu}g/ml$ induced cell death as few mitotic cells were shown. Accordingly, EEPIR ($25-100{\mu}g/ml$) is genotoxic in human lymphocytes and cytotoxic at concentrations of ${\geq}500{\mu}g/ml$ in vitro. Therefore, these activities of the EEPIR could serve its potential therapeutic effects, especially as an anticancer agent. Further study of EEPIR in vivo is now needed to support this in vitro evidence.

Expression of Gas6 Receptors, Tyro3, Axl, and Mertk, in Oocytes and Embryos and Effects of Mertk RNAi on the Oocyte Maturation

  • Kim, Kyeoung-Hwa;Lee, Sang-Eun;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.16 no.3
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    • pp.195-204
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    • 2012
  • Recently, we reported growth arrest-specific gene 6 (Gas6) as a new maternal effect gene (MEG), that expressed in the oocytes but functioned principally during embryogenesis. Gas6 RNAi-treated oocytes developed to metaphase II (MII) stage but they have affected M-phase promoting factor (MPF) activity and incurred abnormal pronuclear (PN) formation during fertilization. Gas6 is a ligand of TAM family members (Tyro3, Axl and Mertk) of receptor tyrosine kinase (RTK). Purpose of the present study was to evaluate the expression of Tyro3, Axl and Mertk transcripts in oocytes and early embryos. Expression of Gas6 and Mertk mRNA was detectable in oocytes and follicular cells, while Tyro3 and Axl mRNA was expressed only in follicular cells. Expression of Mertk mRNA was relatively constant during oocytes maturation and embryogenesis, but the other receptors, Tyro3 and Axl, were not expressed in oocytes and PN stage of embryos at all. Knockdown of Mertk mRNA and protein by using sequence-specific Mertk double strand RNA (dsRNA) did not affect oocytes maturation. In this case, however, contrary to the ligand Gas6 RNA interference (RNAi), MPF activity had not been changed by Mertk RNAi. Therefore, we concluded that the Gas6-Mertk signaling is not directly related to the oocyte maturation. It is still required to study further regarding the function of Mertk as the receptor of Gas6 during preimplantational early embryogenesis.

Development of Bovine Nuclear Transfer Embryos Using Life-span Extended Donor Cells Transfected with Foreign Gene

  • Hwang, Seongsoo;Choi, Eun Joo;You, Seungkwon;Choi, Yun-Jaie;Min, Kwan-Sik;Yoon, Jong-Taek
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1574-1579
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    • 2006
  • This study was performed to determine the developmental potentials of nuclear transfer (NT) embryos using life-span extended cells transfected with a foreign gene as donor cells. A life-span extended bovine embryonic fibroblast cell line was transfected with an expression vector in which the human type II collagen (BOMAR) and ear fibroblasts were used as a donor cell. Cytogenetic analysis was performed to analyze the chromosomal abnormality of donor cells. The fusion rate of 1.8 kV/cm for $15{\mu}sec$ given twice was significantly higher than that of other groups (p<0.05) and the embryos lysed were significantly higher after 1.8 kV/cm for $20{\mu}sec$ given once compared to other groups (p<0.01). The blastocyst development in the ear cell group was statistically significant compared to both BOMAR groups (p<0.01). Both BOMAR groups cultured more than 40 passages (>40 passages) had a lower number of chromosomes; however, fresh granulosa cell (GC) and BOMAR groups cultured less than 20 passages had normal chromosome numbers. Both >40 passages BOMAR groups had numerous obscure debris in metaphase spreads. The transfected foreign gene was expressed in all BOMAR groups, but not in the GC group. Based on these results, the lower developmental potential of NT embryos using life-span extended donor cells transfected with a foreign gene might be a cause of chromosomal abnormality in donor cells.

The effects of different types of media on in vitro maturation outcomes of human germinal vesicle oocytes retrieved in intracytoplasmic sperm injection cycles

  • Fesahat, Farzaneh;Firouzabadi, Razieh Dehghani;Faramarzi, Azita;Khalili, Mohammad Ali
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.2
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    • pp.79-84
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    • 2017
  • Objective: Optimizing in vitro maturation (IVM) media to achieve better outcomes has been a matter of interest in recent years. The aim of this prospective clinical trial was to investigate the effects of different media on the IVM outcomes of immature oocytes at the germinal vesicle (GV) stage. Methods: A total of 400 immature oocytes at the GV stage with normal morphology were retrieved from 320 infertile women aged $31{\pm}4.63years$ during stimulated intracytoplasmic sperm injection (ICSI) cycles. They were divided into groups of homemade IVM medium (I, n = 100), cleavage medium (II, n = 100), blastocyst medium (III, n = 100), and Sage IVM medium (IV, n = 100) and cultured for 24 to 48 hours at $37^{\circ}C$. ICSI was performed, and the rates of fertilization and embryo formation were compared across the four groups. Results: In the 400 retrieved GV oocytes, the total maturation rates showed significant differences in groups I to IV (55%, 53%, 78%, and 68%, respectively, p<0.001). However, there were no significant differences in the fertilization, embryo formation, or arrest rates of metaphase II oocytes across these groups. In all groups, GV maturation was mostly completed after 24 hours, with fewer oocytes requiring 48 hours to mature (p<0.01). Moreover, the rate of high-quality embryos was higher in group IV than in the other groups (p=0.01). Conclusion: The quality of the IVM medium was found to affect clinical IVM outcomes. Additionally, blastocyst medium may be a good choice in IVM/ICSI cycles as an alternative IVM medium.

FISH Karyotype Analysis of Four Wild Cucurbitaceae Species Using 5S and 45S rDNA Probes and the Emergence of New Polyploids in Trichosanthes kirilowii Maxim

  • Waminal, Nomar Espinosa;Kim, Hyun Hee
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.869-876
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    • 2015
  • Wild relative species of domesticated crops are useful genetic resources for improving agronomic traits. Cytogenetic investigations based on chromosome composition provide insight into basic genetic and genomic characteristics of a species that can be exploited in a breeding program. Here, we used FISH analysis to characterize the ploidy level, chromosome constitution, and genomic distribution o f 5S and 4 5S r ibosomal DNA (rDNA) in four wild Cucurbitaceae species, namely, Citrullus lanatus (Thunb.) Mansf. var. citroides L. H. Bailey (2n = 22), Melothria japonica Maxim. (2n = 22), Sicyos angulatus L. (2n = 24), and Trichosanthes kirilowii Maxim. (2n = 66, 88, 110 cytotypes), collected in different areas of Korea. All species were diploids, except for T. kirilowii, which included hexa-, octa-, and decaploid cytotypes (2n = 6x = 66, 8x = 88, and 10x = 110). All species have small metaphase chromosomes in the range of $2-5{\mu}m$. The 45S rDNA signals were localized distally compared to the 5S rDNA. C. lanatus var. citroides and M. japonica showed one and two loci of 45S and 5S rDNA, respectively, with co-localization of rDNA signals in one M. japonica chromosome. S. angulatus showed two co-localized signals of 5S and 45S rDNA loci. The hexaploid T. kirilowii cytotype showed five signals each for 45S and 5S rDNA, with three being co-localized. This is the first report of hexaploid and decaploid cytotypes in T. kirilowii. These results will be useful in future Cucurbitaceae breeding programs.