• Title/Summary/Keyword: cytogenetic study

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Cytogenetic Study of Diploid and Triploid Marine Medaka, Oryzias dancena (해산송사리, Oryzias dancena 유도 3배체의 세포유전학적 연구)

  • Park, In-Seok;Gil, Hyun Woo;Lee, Tae Ho;Nam, Yoon Kwon;Ko, Min Gyun;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.28 no.4
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    • pp.215-222
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    • 2016
  • Triploidy was induced in the marine medaka, Oryzias dancena by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 30, 45, or 60 min, applied two minutes after fertilization. The triploid genotype was induced by each of the thermal shock regimes tested. The best result was obtained when the eggs were treated for 45 min, which induced triploidy in all the resulting fish. Triploidy was confirmed using chromosomal and flow cytometer analyses, and erythrocyte measurements. The surface areas and volumes of the erythrocytes of triploid fish were significantly larger than those of diploid fish, and their chromosome number (3N=72) was 1.5 times greater that for the diploids (2N=48). Based on a flow cytometer analysis, the triploid fish had approximately 1.5 times the cellular DNA content (2.40 pg/cell) of the diploid specimens (1.61 pg/cell). Data from this study provide the basis for the development of unique models for studying reproductive confinement in transgenic fish.

A Study on Environmental Monitoring of Fluorouracil and Decontamination Reagents (Fluorouracil의 환경감시 및 제거약제에 관한 연구)

  • Lee, Su-Gil;Lee, Nae-Woo
    • Journal of the Korean Society of Safety
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    • v.24 no.4
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    • pp.47-52
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    • 2009
  • This study has been to examine the occupational exposure levels of Fluorouracil (5-FU) in a hospital and to investigate the most effective cleaning reagent for control. Fluorouracil is one of the cytotoxic drugs which are therapeutic agents used to treat cancer. The health practitioners working in the cytotoxic work room and oncology ward areas are exposed to adverse health risks like cytogenetic and DNA damage from cytotoxic drugs exposure by frequent skin contact from contaminated surfaces. Four kinds of cleaning reagents has been examined to degrade the 5-FU. It was found that 5-FU was only degraded soon after the reaction in 0.5%(w/v) NaClO solution. Therefore, 0.5%(w/v) NaClO solution has been chosen to decompose any residues on the contamination surfaces. A substantial level of contamination was found on the surfaces of cytotoxic work room and oncology ward areas. The contamination ranges of the surfaces in cytotoxic work room and oncology ward areas were from 2.0 to $13.8{\mu}g/m^2$ and 5.39 to $11.53{\mu}g/m^2$ respectively. Consequently, regulation of the occupational exposure limit, procedure of special cleaning, and the use of personal protective equipment are recommended during the manipulation and administration of the drugs to avoid skin contamination from cytotoxic drugs like 5-FU.

Karyotype Analysis and Physical Mapping of rDNAs Using Bicolor-FISH in Tiarella polyphylla D. Don (헐떡이풀의 핵형분석과 Bicolor FISH를 이용한 물리적 지도 작성)

  • Kim, Soo-Young;Lee, Joong-Ku
    • Korean Journal of Plant Resources
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    • v.20 no.5
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    • pp.446-450
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    • 2007
  • Tiarella polyphylla D. Don(Saxifragaceae) is a perennial herb and distributed in China, Japan, Taiwan and Korea. Especially, it only grows in Ulleung island of Korea. It has been using for asthma, bruise and audition troubles with main components of some Triterpenoids and seven oleanolic Saponins. There is only known its chromosomal number rarely and cytogenetic study was not done. From this study, karyotype analysis and chromosomal localization of 5S and 45S rDNAs using bicolor-FISH(fluorescence in situ hybridization) were carried out. The somatic metaphase chromosome number was 2n=2x=14 and the size of chromosomes ranged $1.66{\sim}3.50{\mu}m$. The chromosome complement consisted of four pairs of submetacentrics(chromosomes 1, 2, 3 and 6), two pairs of subtelocentrics(chromosomes 5 and 7) and one pair of telocentrics(chromosome 4). We also observed NOR(nucleolus organizer region) on the chromosome 4. In bicolor-FISH, one pair of 55 and 45S rDNA sites was detected on the centromeric region of chromosome 3 and short arm of chromosome 4, respectively. Bicolor FISH was very useful tool for the localization and identification of rDNAs on the chromosomes in Tiarella polyphylla.

Utility of Real Time RT-PCR for the Quantitative Detection of Minimal Residual Disease in Hematological Malignancy (백혈병 미세잔존질환 정량검출을 위한 실시간 역전사중합효소연쇄반응법의 유용성)

  • Cho, Jeung-Ai;Kim, Da-Woon;Jeong, Seong-Du;Cheon, Ji-Seon;Na, Gyeong-Ah;Kim, Hye-Ran;Kim, Jin-Gak;Kim, In-Hwan;Kim, Soo-Hyun;Shin, Myung-Geun;Kim, Hyeong-Rok
    • Korean Journal of Clinical Laboratory Science
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    • v.41 no.1
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    • pp.11-23
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    • 2009
  • Chromosomal rearrangements are major pathology in hematological malignancies. The detection of minimal residual disease (MRD) for these gene rearrangements helps in monitoring treatment outcomes and predicting prognosis of patients. Recently, quantification of these gene transcripts based on real-time quantitative polymerase chain reaction (RQ-PCR) has been used as MRD detection. The purpose of this study is to ensure the usefulness of the RQ-PCR technique for detecting MRD in hamatological malignancy patients. The patients had been diagnosed to AML1-ETO positive AML, PML-RARa positive AML and BCR-ABL positive MPN at Chonnam National University Hwasun Hospital from Jan. 2006 to Aug. 2008. The fusion transcript was quntified by RQ-PCR and analyzed in comparison to conventional cytogenetics, FISH and RT-PCR. The fusion gene transcript was quantified by RQ-PCR in 57 samples from 14 patients with AML1-ETO positive AML, 79 samples from 27 patients with PML-RARa positive AML and 108 samples from 36 patients with CML. At diagnosis, the quantitative fusion transcripts for AM1-ETO, PML-RARa and BCR-ABL showed the range of 0.485552651~10.82233683 (mean 3.782217131, SD 2.998052348), 0.005300395~0.29267494 (mean 0.056901315, SD 0.080131381) and 0.1293929~12.94826849 (mean 1.701935665, SD 2.200913158). The increase of AML1-ETO fusion gene transcripts preceded morphologic relapse in two patients. Quantification of fusion gene transcripts by RQ-PCR could detected MRD in samples which were negative by in cytogenetic analysis or FISH. Our findings indicated that quantitative analysis of AML1-ETO, PML-RARa and BCR-ABL transcripts by RQ-PCR might be a useful tool for the monitoring of minimal residual disease in hematological malignancies.

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Production of Supermale (YY) and Superfemale $({\Delta}YY)$ Nile Tilapia (Oreochromis niloticus) by Sex Reversal and Chromosome Manipulation I. Induction of Gynogenetic Diploid from XY Female (성전환 및 염색체 공학 기법을 이용한 초수컷(YY) 및 초암컷$({\Delta}YY)$ 나일틸라피아(Oreochromis niloticus) 생산 I. 성전환된 XY 암컷으로부터 자성발생성 이배체 유도)

  • Kim Dong Soo;Choi Yoon Hee;Noh Choong Hwan;Nam Yoon Kwon
    • Journal of Aquaculture
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    • v.8 no.4
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    • pp.295-306
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    • 1995
  • The present study was performed to produce supermales and superfemales in Oreochromis niloticus by combination of induced sex reversal and diploid gynogenesis. More than $95\%$ of female was obtained by oral administration of $17\beta-estradiol$ (480 mg/kg diet) to the eutheroembryonic larvae of this species. The result of progeny tests with XY pseudofemales showed that incidences of male progeny were ranged from $71.4\%\;to\;73.7\%$. p.opo.lion of male from XY pseudofemale was not significantly different (P>0.05) from that of $\chi^2$ (1:3), but it was significantly different (P<0.01) from that of $\chi^2$(1:1) Gynogenetic diploids were produced by applying cold shocks 3 min after insemination to the eggs of XY pseudofemale sperms were genetically inactivated by ultraviolet rays of 4,050 $erg/mm^2$. Fertilization rates of gynogenetic diploids were not different from that of their controls, however, hatching rates and early survival rates were silghtly lower than those of controls. In the cytogenetic studies, there was no difference in the size of cell and nucleus between the gynogenetic diploids and the controls. At 80 days after hatching, frequencies of gynogenetic male were higher than those of gynogenetic female (P<0.01) in their population.

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Prenatal Aneuploidy Detection in Uncultured Amniotic Fluid Interphase Cells by Fluorescence in situ Hybridization (FISH) (형광직접보합법을 이용한 미배양 양수세포에서 산전 이수배수체 확인)

  • Seol, Hye-Won;Ko, Hee-Jung;Song, Nam-Hee;Kim, Sook-Ryoung;Lee, Hwa-Jin;Oh, Sun-Kyung;Park, Joong-Shin;Jun, Jong-Kwan;Yoon, Bo-Hyun;Syn, Hee-Chul;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.3
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    • pp.223-231
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    • 2003
  • Objective: The aim of the present study was to evaluate the clinical efficiency of fluorescent in situ hybridization (FISH) in the prenatal diagnosis of chromosomal aneuploidy. Methods: We reviewed data of 268 cases to identify women undergoing genetic amniocentesis at cytogenetic laboratory, from January 2000 to December 2002. Amniotic fluid was submitted for both rapid FISH on uncultured interphase amniocytes using a commercially available DNA probe for chromosome 13, 18, 21, X, Y and standard karyotyping on cultured metaphase amniocytes. Results from FISH and full karyotype were compared. Results: There were 251 cases (84%) normal and 17 cases (16%) abnormal in FISH results. All 17 cases of trisomy 13, 18, 21 including two cases of mosaicism and sex chromosome aneuploidies which are detected by FISH were confirmed with conventional cytogenetics and there was no false positive result. Twenty two cases had karyotypically proven abnormalities that could not have been detected by the targeted FISH. Conclusion: Interphase FISH analysis of uncultured amniotic fluid cells has been shown to be an effective and reliable technique for rapid fetal aneuploidy screening during pregnancy as an adjunctive test to conventional cytogenetics.

Midtrimester Amniocentesis for Prenatal Diagnosis (산전 진단을 위한 임신 중기 양수 천자)

  • Kim, Sook-Ryung;Choi, Eun-Jung;Kim, Tae-Yoon;Kwon, Kyoung-Hun;Song, Nam-Hee;Oh, Sun-Kyung;Chi, Hee-Jun
    • Journal of Genetic Medicine
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    • v.5 no.2
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    • pp.125-130
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    • 2008
  • Propose: To analyze the indications and cytogenetic results of midtrimester amniocentesis. Material and Methods: This study reviewed 2,523 cases of midtrimester prenatal genetic amniocentesis performed at MizMedi Hospital between January 2000 and December 2007. Results: The most frequent indication for midtrimester amniocentesis was advanced maternal age (45.9%), followed by positive serum markers (29.9%). Chromosomal aberrations were diagnosed in 110 cases (4.4%), for which numerical aberration accounted for 38 cases (34.5%), structural aberration accounted for 65 cases (59.1%), and mosaicism accounted for 7 cases (6.4%). Among the autosomal aberrations, there were 20 cases of trisomy 21 and 8 cases of trisomy 18. With respect to structural aberrations, there were 14 cases of reciprocal translocation and 8 cases of robertsonian translocation. The frequencies of chromosomal aberrations according to the indication were highest in individuals with a family history of chromosome abnormality 14.0% (8/57) followed by previous congenital anomaly 5.9% (2/34). Conclusion: Midtrimester amniocentesis is an effective tool for prenatal diagnosis. Indications such as advanced maternal age, maternal serum markers, and ultrasound are important for predicting abnormal fetal karyotypes.

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Karyotype analysis of cryopreserved mononuclear cells from cord blood (제대혈 단핵세포의 냉동 전.후의 염색체 핵형분석의 실험적 연구)

  • Ku, Ki-Young;Chu, Mi-Ae;Kim, Ji-Yoon;Lee, Kun-Soo
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.55-60
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    • 2008
  • Purpose : The ability to perform chromosome analysis of cryopreserved cord blood mononuclear cells is important for future retrospective studies. We compared the karyotypes of cryopreserved cells with cells before cryopreservation. Methods : One cord blood (CB) sample was obtained from normal healthy volunteer. Karyotype analysis was performed before cryopreservation. After mononuclear cell separation with Ficoll-Hypaque, the mononuclear cells were cryopreserved by programmed controlled-rate freezer and then transferred into the liquid nitrogen ($-196^{\circ}C$) for 3 days. After rapid thawing, cytogenetic analysis was performed as the same method for each sample by different conditions. The samples were divided by three groups. The first group was no culture before cryopreservation, the second group was 72 hours culture before cryopreservation, but no 24 hours culture after thawing and the third group was 72 hours culture before cryopreservation and 24 hours culture after thawing. Results : The chromosome analysis was successful in the second and third groups of CB sample. Conclusion : The successful result from CB samples may suggest the usefulness of long-term cryopreservation for retrospective study in various clinical settings including hematologic malignancies.

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Analysis of the Azoospermia Factor (AZF) Gene on Y Chromosome and Expression Pattern of DAZ Gene in Korean Infertile Men (한국 남성 불임환자에서 Y 염색체상의 AZF Gene에 대한 분석 및 DAZ Gene의 발현 양상)

  • Lee, Ho-Joon;Lee, Hyoung-Song;Song, Gyun-Jee;Byun, Hye-Kyung;Seo, Ju-Tae;Kim, Jong-Hyun;Lee, You-Sik
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.1
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    • pp.57-65
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    • 1997
  • Cytogenetic observations of loss of the distal portion of the Y chromosome long arm were found to be associated with disrupted spermatogenesis. The existence of a gene involved in the regulation of spermatogenesis, the azoospermia factor (AZF), was postulated. In this study, we screened the AZF region including DAZ and DAZH genes and observed the expression pattern of DAZ and DAZH transcript in infertile men with azoospermia and oligospermia by using a sequence-tagged site (STS)-based PCR method. PCR primers were synthesized for 11 STSs that span Yq interval 6, SRY, DAZ, and DAZH, functional DAZ homologue on chromosome 3. Microdeletions were detected in 4/32 (12.5%) azoospermic men and 1/11 (9%) severe oligospermic men. Only 2 of 5 patients had microdeletions of Yq that contained the DAZ gene, whereas the other 3 patients had deletions extending from intervals 5L-6F proximal to the DAZ gene on Yq. Testis biopsies of the azoospermic patients revealed a variety from Sertoli cell-only syndrome to testicular maturation arrest. Of 4 men with clinical data available, average testis size was R: 13.8 cc, L: 13.8 cc, serum T was $4.0{\pm}1.25$ ng/ml, LH was $3.63{\pm}1.90$ mIU/ml, and FSH was $8.85{\pm}5.13$ mIU/ml. These values did not differ significantly from the remainder of the patients tested. We could not observed the DAZ transcript in 2 patients, who have no mature spermatozoa. In 11.6% of patients microdeletions of the AZF could be detected. These deletions in the AZF region seem to be involved causing spermatogenic failure. But the frequency of microdeletions proximal to DAZ suggests that DAZ is not the only gene associated with spermatogenic failure.

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Induction of Radiation Adaptive Response in Lymphocytes of patients Undergoing Bone Scintigraphy (골 신티그라피를 시행한 환자의 림프구에서 방사선 적응반응의 유도에 관한 연구)

  • Min, Jung-Jun;Bom, Hee-Seung
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.2
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    • pp.172-177
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    • 1999
  • Purpose: The purpose of this study was to ascertain whether radiation adaptive response could be induced by Tc-99m-methylene diphosphonate (Tc-99m-MDP) in peripheral lymphocytes of patients undergoing bone scintigraphy. Materials and Methods: Lymphocytes from 22 patients (6 males, 16 females, mean age $50{\pm}14$ years) were collected before and after bone scintigraphy using 740 MBq Tc-99m-MDP Lymphocytes from 10 controls (6 males, 4 females, mean age $43{\pm}7$ years) were also collected. They were exposed to challenge dose of 2 Gy gamma rays using a cell irradiator. Number of ring-form and dicentric chromosomes per 600 cells (chromosomal aberrations) was counted under the light microscope. Results: Chromosomal aberrations in patients before bone scintigraphy ($385.1{\pm}30.5$) was not different from that of controls ($367.8{\pm}36.6$). However, chromosomal aberrations in patients after bone scintigraphy was significantly decreased to $192.6{\pm}22.1$ (p=0.0001). Conclusion: Low dose gamma-irradiation by Tc-99m-MDP used for bone scintigraphy induces a cytogenetic adaptive response in peripheral lymphocytes.

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