• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.405-415
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• 2000

Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.24-24
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• 2003

Fluorescence in situ Hybridization(FISH)는 특정 염기서열을 이용하여 염색체나 염색체상의 DNA위치를 확인하는 기술로서, 면역세포화학 기술과 결합되어져 현미경으로 이들의 유전적 활성도를 직접 확인할 수 있는 방법으로 지금까지의 radioisotopes 대신 non-radioactive labeling 방법으로서 fluorescence을 이용한 분자세포유전학적 검정 방법이다. 따라서 특정 염색체의 FISH probe의 개발은 FISH 기법을 이용하여 조직 또는 세포내 특정 염색체나 DNA의 존재나 이상 유무를 신속하고 정확하게 파악할 수 있다. 본 연구는 소와 사람을 대상으로 Y-염색체 특이 DNA probe를 개발하고 이를 이용하여 FISH를 시행함으로서 본 probe의 신뢰성을 확인하고 임상적 적용 가능성을 제시 하고자 하였다.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.51-56
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• 1997

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'dysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.

• Journal of Genetic Medicine
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• v.3 no.1
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• pp.5-10
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• 1999

This is a case report of 46,XY female phenotype (46,XY karyotype, no pubic hair, blind vagina and absence of uterus)in an 18-year-old patient. To confirm whether a Y chromosome has a structural abnormality, fluorescent in situ hybridization (FISH) with the chromosome X/Y cocktail probe was simultaneously performed, and the six loci [PABY, RPS4Y(sy16, sy17), ZFY, DYS14] on the short arm, one locus (DYZ3) on the centromere and one locus (DYZ1) on the long arm were amplified by polymerase chain reaction (PCR). The probes used FISH hybridized to centromere of the X chromosome and heterochromatin region (Yq12) of the Y chromosome, and all PCR related Y chromosome showed positive band like normal male. From the results obtained, it seemed that the Y chromosome from the 46,XY female was structurely normal. Especially, the SRY gene has been equated with the mammalian testis-determining factor, and absence or point mutation in the SRY gene causes XY female. To detect the point mutations of SRY sequences, single-strand conformation polymorphism (SSCP) assay was used. Our results confirm that this patient has no mutation in the SRY gene on the Y chromosome.

• Journal of Genetic Medicine
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• v.7 no.2
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• pp.160-164
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• 2010

Structural abnormalities of the Y chromosome affect normal testicular differentiation and spermatogenesis. The present case showed a rare monocentric derivative Y chromosome with partial duplication of Yp including the SRY gene and deletion of Yq12 heterochromatin. The karyotype was 46,X,der(Y)(pter${\rightarrow}$q11.23::p11.2${\rightarrow}$pter).ish der(Y)(DYZ3+,DYZ1-,SRY++), confirmed through a FISH study. Even though the patient possessed an abnormal Y chromosome, testicular biopsy showed normal testicular volumes in the proband, with gonadal hormonal levels in the normal range but bilateral varicocele and hypospermatogenesis. We speculate that the abnormal Y chromosome arose from sister chromatids during Y chromosome recombination or intra chromosomal NAHR (non-allelic homologous recombination) during meiosis in the patient's father or in the very early stages of embryogenesis. The derivative Y chromosome might interfere in the meiotic stage of spermatogenesis, leading to the developmental arrest of germ cells. The present case illustrates that the infertility phenotype can have various causes. Also, it emphasizes the importance of accurate and various genetic analyses and could aid in male infertility treatment.

• Bulletin of the Korean Chemical Society
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• v.18 no.9
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• pp.1002-1006
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• 1997

1H NMR spectra of D2O solutions containing 2,2-dimethylaziridine (1) or 2-methylaziridine (2) and [SiW11COⅡO39]6- (SiW11Co) or [SiW11NiⅡO39]6- (SiW11Ni) exhibit separate signals for the free ligand and the complex, indicating that the ligand exchange is slow on the NMR time scale. Identified are two linkage isomers with the methyl group of 2 at trans or cis position with respect to the metal. The isotropic shifts of 1 and 2 coordinated to SiW11Ni originate mainly from the contact shifts, and they agree reasonably with the relative values reported for similar ligands coordinated to bis(2,4-pentanedionato)nickel(Ⅱ). The isotropic shifts for the SiW11Co complexes were separated into contact and pseudocontact contributions. The pseudocontact shifts show that (χ∥-χ⊥) is positive, while that for the SiW11Co complexes of pyridine derivatives is negative. This result indicates that the ordering of dxy and dxz, dyz orbitals in SiW11Co complexes can be reversed by ligands.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.287-287
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• 2004

Sexing from bovine embryos which were fertilized in vitro implicate a possibility of production of the sex controlled cattle. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. (omitted)

• Proceedings of the KSMRM Conference
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• 2001.11a
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• pp.112-112
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• 2001

Purpose： We developed the software for diffusion tensor imaging and evaluated its feasibility in norm brains. Method： Five normal volunteers, aged from 25 to 29 years, were examined on a 1.5 T MR system. the diffusion tensor pulse sequence used a SE-EPI with 6 diffusion gradie directions of (1, 1, 0), (-1, 1,0), (1, 0, 1), (-1, 0, 1), (0, 1, 1), (0, 1, -1) and also with no diffusion gradient. A b-factor of 500 sec/mm2 was used. Measurement parameter were as follows； TR/TE=10000 ms/99 ms, FOV=240 mm, matrix=128$\times$128, slice thickness/gap=6 mm/0 mm, bandwidth=91 kHz and the number of total slices=20. Four repeated axial diffusion images were averaged for diffusion tensor imaging. A total scan 11 of 4 min 30 sec was used. Six full diffusion tensor components of Dxx, Dyy, Dzz, Dxy, Dxz and Dyz were obtained using two-point linear regression model from 7 diffusion-weight images at each pixel and fractional anisotropy and lattice index images was estimated fr their eigenvectors and eigenvalues. Our program was written on a platform of IDL. W evaluated the qualities of fractional anisotropy and lattice index images of normal brains a knew whether our software for diffusion tensor imaging may be feasible.

• Journal of Genetic Medicine
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• v.2 no.1
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• pp.11-15
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• 1998

This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX, and the presence of Y chromosome mosaicism was ruled out through XY dual fluorescent in situ hybridization (FISH). By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.

• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.133-137
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• 2004

Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.