• Toxicological Research
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• v.33 no.4
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• pp.315-323
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• 2017

Preconceptual sex selection is still a highly debatable process whereby X- and Y-chromosome-bearing spermatozoa are isolated prior to fertilization of the oocyte. Although various separation techniques are available, none can guarantee 100% accuracy. The aim of this study was to separate X- and Y-chromosome-bearing spermatozoa using methods based on the viability difference between the X- and Y-chromosome-bearing spermatozoa. A total of 18 experimental semen samples were used, written consent was obtained from all donors and results were analysed in a blinded fashion. Spermatozoa were exposed to different pH values (5.5, 6.5, 7.5, 8.5, and 9.5), increased temperatures ($37^{\circ}C$, $41^{\circ}C$, and $45^{\circ}C$) and ROS level ($50{\mu}M$, $750{\mu}M$, and $1,000{\mu}M$). The live and dead cell separation was done through a modified swim-up technique. Changes in the sex-chromosome ratio of samples were established by double-label fluorescent in situ hybridization (FISH) before and after processing. The results indicated successful enrichment of X-chromosome-bearing spermatozoa upon incubation in acidic media, increased temperatures, and elevated $H_2O_2$. This study demonstrated the potential role for exploring the physiological differences between X-and Y-chromosome-bearing spermatozoa in the development of preconceptual gender selection.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.15-21
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• 1999

The aim of the present study was to determine the effect of paternal sex chromosome on early development of buffalo embryos fertilized and cultured in vitro. Embryos were produced in vitro from abattoir derived buffalo oocytes. The cleaved embryos were cocultured with buffalo oviductal epithelial cells and evaluated on day 7 under the phase contrast microscope to classify development. The embryos which reached the morula/blastocyst stage were fast developing, the embryos which were at 16-32 cell stage were medium developing and the embryos below 16 cell stage were slow developing. The embryos which showed some fragmentation in the blastomeres or degenerated blastomeres, were degenerating. Sex of emberyos (n=159) was determined using PCR for amplification of a male specific BRY. 1 (301 bp) and a buffalo specific satellite DNA (216 bp) fragments. The results thus obtained show that 1) X and Y chromosome bearing sperms fertilize oocytes to give almost equal numbers of cleaved XX and XY embryos, 2) male embryos develop faster than female embryos to reach advanced stage and 3) degeneration of buffalo embryos is not linked with the paternal sex chromosome. We suggest that faster development of males is due to differential processing of X and Y chromosome within the zygote for its activation and / or differential expression of genes on paternal sex chromosome sex chromosome during development of buffalo embryos fertilized and cultured in vitro which may be attributed to a combination of genetic and environmental factors.

• Korean Journal of Medicinal Crop Science
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• v.26 no.2
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• pp.113-118
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• 2018

Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small ($2.71-5.50{\mu}m$)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2003.10a
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• pp.183-192
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• 2003

With the availability of complete whole-genomes such as the human, mouse, fugu and chimpanzee chromosome 22, comparative analysis of large genomes from cross-species at varying evolutionary distances is considered one of a powerful approach for identifying coding and functional non-coding sequences. Here we describe a fast and efficient global alignment method especially for large genomic regions over mega bases pair. We used an approach for identifying all similarity regions by HSP (Highest Segment Pair) regions using local alignments and then large syntenic genome based on the both extension of anchors at HSP regions in two species and global conservation map. Using this alignment approach, we examined rearrangement loci in human chromosome 21 and chimpanzee chromosome 22. Finally, we extracted syntenic genome 30 Mb of human chromosome 21 with chimpanzee chromosome 22, and then identified genomic rearrangements (deletions and insertions ranging h size from 0.3 to 200 kb). Our experiment shows that all jnsertion/deletion (indel) events in excess of 300 bp within chimpanzee chromosome 22 and human chromosome 21 alignments in order to identify new insertions that had occurred over the last 7 million years of evolution. Finally we also discussed evolutionary features throughout comparative analyses of Ka/ks (non-synonymous / synonymous substitutions) rate in orthologous 119 genes of chromosome 21 and 53 genes of MHC-I class in human and chimpanzee genome.

• Reproductive and Developmental Biology
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• v.31 no.4
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• pp.273-278
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• 2007

This study was conducted to examine the protein kinase inhibitors, 6-dimethylaminopurine (DMAP) and cycloheximide (CHXM) on the development and chromosome constitution of porcine parthenogenetic embryos. In vitro matured oocytes were activated by electric stimuli (ES) or a combination of ES with culture in 2 mM DMAP or $10{\mu}g/ml$ CHXM for 4 hr. Activated oocytes were cultured in PZM-3 for 6 days. Some 1-cell embryos and blastocysts were fixed by air dry method to analyze the chromosome constitutions and/or total cell number. Blastocyst development of DMAP-treated group (26.7%) was significantly higher (p<0.05) than those of CHXM-treated and ES control groups. Ploidy in 1-cell stage embryos was not different among groups (77.3 to 81.0%), however, proportion of diploid chromosome constitutions was high in DMAP-treated group (61.9%, p<0.05). In the blastocyst stage, proportion of diploid chromosome plates was significantly high in DMAP-treated group (64.2%, p<0.05), and proportion of abnormal chromosome plates was higher in CHXM-treated group (36.6%, p<0.05) than DMAP-treated group (28.3%,). Proportion of embryos with abnormal chromosome constitutions was slightly increased by DMAP (40.0%) and CHXM (42.1%) treatment due to the increasing of mixoploid (47.4 and 52.0%). The present study shows that the DMAP treatment increase the development of porcine parthenotes. However, parthenogenetic activation by ES or combined treatment with ES and DMAP or CHXM detrimentally affects the chromosome constitutions of porcine parthenotes during early embryonic development, leads to increased abnormal ploidy in the blastocyst stage.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.467-474
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• 1999

Objectives: Cytogenetic investigations were carried out on 770 women with primary (n=560) and secondary amenorrhea (n=210) to determine the frequency of chromosomal or genetic causes of amenorrhea. Materials and Methods: In 770 women with primary amenorrhea (n=560) and secondary amenorrhea (n=210), chromosomal analysis were performed. Results: 1) The most prevalent age group is 16-20 years of age group with primary amenorrhea and 26-30 years of age group with secondary amenorrhea. 2) Out of 560 cases of primary amenorrhea, 343 cases (61.3%) had the normal chromosome constitution and 217 cases (38.7%) had the abnormal chromosome constitution including 46,XY. 3) In 217 cases of abnormal chromosome of primary amenorrhea, 57 cases (26.3%) had 45,X and 34 cases (15.8%) had the 46,XY, 24 cases (11.0%) had 45,X/46,X,i (Xq), 23 cases (10.6%) had 45,X/46,X,+mar and 14 cases (6.6%) had 45,X/46,XY. 4) Out of 210 cases of secondary amenorrhea, 181 cases (86.2%) had the normal chromosome constitution and 29 cases (13.8%) had the abnormal chromosome. 5) In 29 cases of abnormal chromosome of secondary amenorrhea, 7 cases (24.1%) had 45,X/46, X,i (Xq), 4 cases (13.8%) had 45,X/46,XX. Conclusion: High percentage of chromosomal abnormalities was diagnosed in primary amenorrhea and most of them were sex chromosome anomalies. In secondary amenorrhea, the prevalence was lower than primary amenorrhea, so a preselection of patients with secondary amenorrhea for cytogenetic investigations seems to be necessary.

• Korean Journal of Plant Taxonomy
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• v.40 no.3
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• pp.153-156
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• 2010

The somatic chromosome number of four Korean species of Leontopodium were investigated. The chromosome number of L. leiolepis (2n = 24) is reported here as for the first time. The chromosome number of L. japonicum (2n = 28) is not varied among the 3 populations on the Korean Peninsula, but that condition is different from the previous reports for Korea (2n = 26) and Japan (2n = 21, 26). L. hallaisanense and L. japonicum, both of which are in sect. Nobilia and similar to each other in gross morphology, have the same chromosome number of 2n = 28. On the other hand the chromosome number of Korean L. leontopodioides (2n = 24) is different from that in Russian reports (2n = 26). The chromosome numbers of all Korean species of the genus Leontopodium could be inferred as tetraploid or aneuploid.

• Journal of Radiation Protection and Research
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• v.26 no.2
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• pp.101-111
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• 2001

To study the relationship between the DNA content of human chromosomes and their involvement in radiation-induced structural chromosome aberrations, the frequencies of translocations and dicentrics induced in human lymphocytes after in vitro exposure to 2Gy of x-tay were analysed by fluorescence in situ hybridization(FISH). Single whole chromosome probes(WCP), specific for chromosomes 1, 2, 4, 7, 8, 9, and 21 and triple combination of probes tot chromosome 1, 2 & 4 were used separately. A significant deviation of the frequency of symmetrical translocation and dicentrics from a DNA-proportional distribution was observed. Chromosomes 2, 7, 8, 9 and 21 were less frequently involved in the formation of symmetrical translocations and dicentrics than expected, whereas chromosomes 1 and 4 were more frequently involved. Chromosome 2 and 4 showed a higher frequency of acentric fragments. When triple combination probe for chromosome 1, 2 & 4 was used, no differences were found between the observed and expected frequency of exchange type aberrations. The results showed that the frequency of radiation-induced chromosome aberrations was not proportional to DNA contents, suggesting the difference in the susceptibility to specific aberrations among individual chromosomes. The results also indicated that the FISH technique with combination of probes for chromosome 1, 2 & 4 was useful for radiation biodosimetry.

• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.317-331
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• 1990

Chromosomes of gonadal tissues from Fasciola spp, Eurytrema pancreaticum and Calicophoron calicophorum occurred Korean cattle were egamined using modified air-drying method. To compare their phenotype with three different genotypes among Fasciola spp, the adult and egg si2e were measured since they have been known as important taxonomical characters. The results obtained were as followed; Cattle liver fluke, Fasciola spp were classified into three types based on their chromosomal complements such as individual with 2o chromosome(diploid), 30 chromosome(triploid) and 20/30 mosaic constitution(mixoploid). The propotions of appearance of three types were 40.00%, 54.29% and 5.71%, respectively. The frequency of three types in type I which was regarded as F gigantica were 58.82% for diploid, 35.29% for triploid and 5.88% for mixoploid, but in type II which was regarded as F hepatica were 72.2% for triploid, 22.22% for diploid and 5.56% for mixoploid. Egg length of triploid forms was significantly larger than that of diploid forms and egg size of mixoploid forms was similiar to that of triploid forms. Worm size of triploid forms was larger than that of diploid forms and was more similar to that of mixoploid forms, but the statistical data were not significant. Diploid chromosome consisted of one pair of metacentric chromosome(m), four pairs of submetacentric chromosomes(sm), five pairs of subtelocentric chromosomes(st), while triploid chromosome consisted of one pair of metacentric chromosome, seven pairs of submetacen.tric chromosomes, one pair of subtelocentric chromosome and telocentric chromosome(t), respectively. In mixoploid chromosome, constitution of the chromosomes of diploid or triploid cell was consistent with that of diploid or triploid. Chromosomes of gonadal tissues from pancreatic fluke, Eurytrema pancreaticum consisted of 13 pairs of homologs(2n=26, n=13). The mitotic and meiotic divisions were observed frequently. In the mitotic metaphase, Karyotype consisted of five pairs of metacentric chromosomes, four pairs of submetacentric chromosomes, three pairs of subtelocentric chromosomes and one pair of telocentric chromosome. Chromosomes of gonadal tissues from stomach fluke, Calicvphoron calicophorum consisted of 9 pairs of homologs(2n=18, n=9). The meiotic divisions was frequently observed, but mitotic divisions was rare. In the mitotic metaphase, Karyotype consisted of two pairs of metacentric chromosomes, three pairs of submetacentric chromosomes and four pairs of subtelocentric chromosomes. Karyotype of Calicophoron calicophorum differed from that of Japanese C calicophorum which was similar to that of Paramphistomum cervi of Korean cattle. Though that of Calicophoron calicophorum of Korean cattle was similar to that of Paramphistomum explanatum of Korean cattle, that have been recognized to be a different species of fluke.

• Journal of Plant Biology
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• v.14 no.4
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• pp.14-18
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• 1971

Lilium Miquelianum Makino is a species which originated in Korea. The Karyotype of the species was examined in materials collected at Mts. Kaya, Kasan, Chejung, and Kaji. The results are as follows: 1) The somatic chromosome number was found to be 2n=24. 2) The karyotype is described as: K=2Am＋2Bkm＋2Csst＋2Dsst＋2Ests＋2Fst＋2Gst＋2Hst＋2Ist＋2Jsst＋2Kst＋2Lst m: metacentric, sm: submetacentric st: subtelocentric, s: secondary constriction 3) A single subcentric supernumerary B-chromosome was found in some bulbs from Mt. Kasan. 4) The shape of the supernumerary B-chromosome was similar to that of the E chromosome which had separated at its secondary constriction and lost its lower chromosome fragment. 5) From three to eight nucleio of varying sizes were found in the telophase or interphase nucleus of root tip cells. The maximum number of eight nucleoli corresponds to the number of chromosomes that have a secondary constriction.