• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.179-184
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• 2001

We evaluated a possibility of the use of chloroplast number per guard cell pairs as a measure for ploidy level in the different ploidy levels of tobacco plant (Nicotiana tabacum L. cv. BY-4) . The guard-cell chloroplast numbers of leaves of haploid plant were a half of wild-type plant. Furthermore, the number of chloroplast per guard cell pairs of the leaves of doubled-haploid plant increased in two times compared with that of haploid plant. In addition, the chloroplast number was not changed in the F$_1$ progenies. The change of the chloroplast number by leaf stage was not observed. The results indicate that there is a strong relationship between ploidy level (2x and 4x) and chloroplast number per guard cell pairs. This relationship was also, observed in both in vitro and pot cultured plants. It was determined that the measurement of chloroplast number in guard cells of leaf epidermis is simple to use and less labour intensive, and hence can be considered a practical alternative to the chromosome counting methods or flow cytometry in the tobacco plant.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.129-138
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• 2002

Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.1
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• pp.17-20
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• 2001

Cytogenetic characteristics of three species in Centropomidae (Coreoperca herzi, C. kawamebari and Siniperca schezeri) were evaluated, based on karyologcial analysis, erythrocytic measurement and genome size estimation using flow cytometry. Modal chromosome number of three species was same as 2n=48. Karyotypes were 4SM+44A$\cdot$T (NF=52) for Coreoperca herzi, 6SM+42 A$\cdot$T (NF=52) for C. kawamebari and 4SM+44A$\cdot$T (NF=52) for Siniperca schezeri. Heteromorphic sex chromosome was not found in both sexes of any species examined, Cellular and nuclear volumes of Siniperca schezeri were smaller than those of other two species, Average amounts of cellular DNA contents estimated by flow cytometry were well coincided with erythrocytic sizes. The estimated genome sizes were 1.83, 1.85 and 1.44 pg/cell for C. herzi, C. kawamebari and S. schezeri, respectively.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7343-7350
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• 2015

Genetic changes associated with acute lymphoblastic leukemia (ALL) provide very important diagnostic and prognostic information with a direct impact on patient management. Detection of chromosome abnormalities by conventional cytogenetics combined with fluorescence in situ hybridization (FISH) play a very significant role in assessing risk stratification. Identification of specific chromosome abnormalities has led to the recognition of genetic subgroups based on reciprocal translocations, deletions and modal number in B or T-cell ALL. In the last twelve years 102 newly diagnosed childhood/adult ALL bone marrow samples were analysed for chromosomal abnormalities with conventional G-banding, and FISH (selected cases) using specific probes in our hospital. G-banded karyotype analysis found clonal numerical and/or structural chromosomal aberrations in 74.2% of cases. Patients with pseudodiploidy represented the most frequent group (38.7%) followed by high hyperdiploidy group (12.9%), low hyperdiploidy group (9.7%), hypodiploidy (<46) group (9.7%) and high hypertriploidy group (3.2%). The highest observed numerical chromosomal alteration was high hyperdiploidy (12.9%) with abnormal karyotypes while abnormal 12p (7.5%) was the highest observed structural abnormality followed by t(12;21)(p13.3;q22) resulting in ETV6/RUNX1 fusion (5.4%) and t(9;22)(q34.1;q11.2) resulting in BCR/ABL1 fusion (4.3%). Interestingly, we identified 16 cases with rare and complex structural aberrations. Application of the FISH technique produced major improvements in the sensitivity and accuracy of cytogenetic analysis with ALL patients. In conclusion it confirmed heterogeneity of ALL by identifying various recurrent chromosomal aberrations along with non-specific rearrangements and their association with specific immunophenotypes. This study pool is representative of paediatric/adult ALL patients in Oman.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.105-113
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• 2011

The present study investigated the nuclear remodeling, development potential with telomerase activity and transcription level of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos using two different fusion and activation methods. Female adult fibroblasts were injected into perivitelline space of in vitro matured oocytes. The oocyte-nucleus complexes were fused and followed by immediately either activated (Group 1), or activated at 1 h post-fusion (hpf) (Group 2), respectively. The incidence of normal premature chromosome condensation (PCC) at 1 hpf was slightly increased in the Group 2, compared to those of Group 1, but there was no significant (p<0.05) difference. The incidence of normal pronucleus (PN) and chromosome spread at 5 and 18 hpf were significantly (p<0.05) higher in the Group 2 than those of Group 1. The cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cell numbers were significantly (p<0.05) higher in the Group 2, compared to those of Group 1. Level of telomerase activity was significantly (p<0.05) higher in the SCNT blastocysts of Group 2, compared to those of Group 1. Transcript levels of HPRT, MeCP2 and XIST were not significantly (p<0.05) different between blastocysts of Group 1 and 2. However, transcript level of ANT3, RPS4X, XIAP and ZFX were significantly (p<0.05) up-regulated in the SCNT blastocysts of Group 2, compared to those of Group 1. Taken together, it is concluded that oocyte activation at 1 hpf induces the enhanced developmental potential by efficient nuclear remodeling and subsequent facilitation of the nuclear reprogramming of bovine SCNT embryos.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.26-26
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• 2003

Hyaluronic acid (HA)-binding sites have been shown the diagnostic potential fur assessment of sperm maturity, which is related to male fertility. This study was designed to evaluate chromosomal patterns in porcine embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with non- or HA-binding sperm (HABS). For binding of sperm with HA, sperm incubated in 10 ${mu}ell$ drop containing HA (0.8 mg/ml)-agarose (0.8%) mixture for 15 min. IVF and ICSI with non- or HA-bound sperm examined with matured oocytes at 44 hr after in vitro maturation. Embryos were cultured in 50 ${mu}ell$ of NCSU 23 containing 0.5% BSA for 5 days and then in 50 ${mu}ell$ of NCSU 23 containing 10% FBS for 2 days. For the evaluation of chromosomal aneuploidies, chromosome 1 sub-metacentric specific probe was used in sperm and embryos by fluorescence in situ hybridization (FISH). The frequency of aneuploidy sperm for chromosome 1 was 6.25%. The significant differences following IVF and ICSI with non- or HA-bound sperm were not observed in blastocyst formation rates (18.6, 23.5, and 23.8%) and cell number (61.8 $\pm$ 12.5, 55.5 $\pm$ 7.3, and 59.3 $\pm$ 9.6). Moreover, the percentage of diploidy in 4-cell stage embryos was 57.1% (IVF), 68.8% (ICSI), and 76.3% (ICSI-HABS). These results suggest that HA-binding sites may be a material for selection of normal sperm for ICSI. Therefore HA selection of normal sperm may be reduce the loss to embryonic mortality prior to embryo transfer in pig.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.17-21
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• 2012

The purpose of this study was to improve mapping power and resolution for the QTL influencing carcass quality in Hanwoo, which was previously detected on the bovine chromosome (BTA) 6. A sample of 427 steers were chosen, which were the progeny from 45 Korean proven sires in the Hanwoo Improvement Center, Seosan, Korea. The samples were genotyped with the set of 2,535 SNPs on BTA6 that were imbedded in the Illumina bovine 50 k chip. A linkage disequilibrium variance component mapping (LDVCM) method, which exploited both linkage between sires and their steers and population-wide linkage disequilibrium, was applied to detect QTL for four carcass quality traits. Fifteen QTL were detected at 0.1% comparison-wise level, for which five, three, five, and two QTL were associated with carcass weight (CWT), backfat thickness (BFT), longissimus dorsi muscle area (LMA), and marbling score (Marb), respectively. The number of QTL was greater compared with our previous results, in which twelve QTL for carcass quality were detected on the BTA6 in the same population by applying other linkage disequilibrium mapping approaches. One QTL for LMA was detected on the distal region (110,285,672 to 110,633,096 bp) with the most significant evidence for linkage (p< $10^{-5}$). Another QTL that was detected on the proximal region (33,596,515 to 33,897,434 bp) was pleiotrophic, i.e. influencing CWT, BFT, and LMA. Our results suggest that the LDVCM is a good alternative method for QTL fine-mapping in detection and characterization of QTL.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.1
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• pp.1-8
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• 2005

Myotonic Dystrophies type 1 and 2 (DM1/2) are neuromuscular disorders which belong to a group of genetic diseases caused by unstable CTG triplet repeat (DM1) and CCTG tetranucleotide repeat (DM2) expansions. In DM1, CTG repeats are located within the 3' untranslated region of myotonin protein kinase (DMPK) gene on chromosome 19q. DM2 is caused by expansion of CCTG repeats located in the first intron of a gene coding for zinc finger factor 9 on chromosome 3q. The CTG and CCTG expansions are located in untranslated regions and are expressed as pre-mRNAs in nuclei (DM1 and DM2) and as mRNA in cytoplasm (DM1). Investigations of molecular alterations in DM1 discovered a new molecular mechanism responsible for this disease. Expansion of un-translated CUG repeats in the mutant DMPK mRNA disrupts biological functions of two CUG-binding proteins, CUGBP and MNBL. These proteins regulate translation and splicing of mRNAs coding for proteins which play a key role in skeletal muscle function. Expansion of CUG repeats alters these two stages of RNA metabolism in DM1 by titrating CUGBP1 and MNBL into mutant DMPK mRNA-protein complexes. Mouse models, in which levels of CUGBP1 and MNBL were modulated to mimic DM1, showed several symptoms of DM1 disease including muscular dystrophy, cataracts and myotonia. Mis-regulated levels of CUGBP1 in newborn mice cause a delay of muscle development mimicking muscle symptoms of congenital form of DM1 disease. Since expansion of CCTG repeats in DM2 is also located in untranslated region, it is predicted that DM2 mechanisms might be similar to those observed in DM1. However, differences in clinical phenotypes of DM1 and DM2 suggest some specific features in molecular pathways in both diseases. Recent publications suggest that number of pathways affected by RNA CUG and CCUG repeats could be larger than initially thought. Detailed studies of these pathways will help in developing therapy for patients affected with DM1 and DM2.

• Journal of Ginseng Research
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• v.33 no.1
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• pp.65-71
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• 2009

This experiment was done to determine the optimum conditions for the induction of tetraploidy in Panax ginseng C. A. Meyer using bud length, temperature and plant growth regulator pretreatments. Highest callus formation was obtained when the medium was inoculated with flower bud in the size of 2-3 mm in length. The optimum temperature for the callus formation was high when treated at $4^{\circ}C$ for 4-5 days. Among the treatments of growth regulators and different concentration, highest callus formation was observed in combination of 5 mg/L 2,4-D and 1 mg/L kinetin for P. ginseng. As a result of flow cytometer analysis, all 7 adventitious roots were confirmed as tetraploidys. Cytological analysis revealed that the chromosome number of tetraploid roots was 96, while that of diploid roots was 48. Tetraploid ginseng roots were inoculated to flower bud size of 2-3 mm in length. The callus formation was optimum when treated with 1 mg/L 2,4-D at $4^{\circ}C$ for 5 days. Compared with control roots, tetraploid roots were thicker and longer and had few lateral branches. Fresh weight of tetraploid roots was relatively higher than the control roots.