• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.112-119
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• 2005

Efficient plant regenerationsystem from cell suspension cultures was established in D. acicularis (2n = 90) by monitoring ploidy level and visual selection of the cultures. The highly regenerable cell lines selected maintained original ploidy level and consisted of compact cell clumps with yellowish color and relatively moderate growth, suggesting that it is possible to select visually the highly regenerable cell lines with the original ploidy level. All the regenerated plantlets from the highly regenerable cell cultures exhibited normal phenotypes and no variations in ploidy level were observed by flow cytometry (FCM) analysis.

• 대한수의학회지
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• 제32권4호
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• pp.649-661
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• 1992

This study was performed to compare DNA content by flow cytometer (FCM) and glutathione S-transferase placental form (GST-P) positive foci for searching objective and accurated properties of tumor. Sprague-Dawley rats aged six weeks were divided into three groups and group 1 and 2 of rats were given an intraperitoneal injection of diethylnitrosamine at 200mg/kg body weight and group 3 of rats were given saline. Three weeks after beginning of the experiment, all groups were performed partial hepatectomy. Group 1 of rats were begun to feed on diets containing 0.02% 2-acetylaminofluorene as a promoter for six weeks, group 2 and 3 of rats were begun to feed on basal diets. At 4, 6, and 8 weeks after initiation, all groups of rats were killed, livers were extracted for H & E stain, immunohistochemical stain, and DNA ploidy analysis. In quantitative analysis for GST-P positive lesion number and area by using Image Analyzer, group 1 and 2 represented significant difference in comparison with group 3. In ploidy distribution, diploid cells of group 1 and 2 were increased significantly in comparison with those of group 3 at 4, 6, and 8 weeks after initiation, respectively tetraploid cells were reduced. But S-phase cells were not changed significantly. It is concluded that ploidy change by FCM is useful as objective data for early detection in hepatocarcinogenesis. Therefore, methodology and study of DNA content are carried out for more objective and accurate ploidy analysis in liver tumor.

• Tuberculosis and Respiratory Diseases
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• 제42권3호
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• pp.314-321
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• 1995

연구배경: 종양세포는 세포의 비정상적인 분열성장이 증가되므로, 세포내의 DNA가 양적인 변화를 일으킨다. DNA의 양적변화인 DNA ploidy 여부는 종양의 생물학적 특성을 반영하므로, 소세포 폐임에서 DNA ploidy의 변화와 생존기간을 비교하였다. 방법: 1990년 1월부터 1991년 12월까지 원광의대 부속병원에서 원발성 소세포 폐암으로 조직병리학적 진단을 받고나서, 2회 이상의 화학요법을 실시받은 후, 최소 2년이상의 후향적 추적에 의해 사망이 확인된 42례를 대상으로 하였다. DNA ploidy 분석방법은 paraffin에 보관된 병리조직을 이용하여 유식세포 분석법에 의한 DNA histogram으로 분석하였다. DNA ploidy 여부와 평균 생존기간을 비교하였으며, 다시 TNM 병기, PS scale, 화학요법 실시 횟수 등으로 세분하여 DNA ploidy 여부와 생존기간과의 관계를 재비교하였다. 결과: 1) 전 군의 평균 생존기간은 190(${\pm}156$)일이었으며, TNM 병기, PS scale이 진행할 수록 생존기간은 단축되었다. 2) 전 군에서 DNA aneuploidy의 발현 비율은 60%(26/42)였으며, 암의 진행정도와는 관계없었다. 3) 전 군에서의 평균 생존기간은 diploidy군이 272(${\pm}197$)일로서 aneuploidy 군의 138(${\pm}90$)일보다 유의하게 연장되었다(p<0.001). 4) DNA ploidy 여부에 의한 생존기간의 차이에 대한 TNM 병기, PS scale의 영향은 없었다. 결론: 소세포 폐암 환자에서 DNA aneuploidy 군은 diploidy 군보다 유의하게 생존기간이 짧았으며, DNA ploidy 여부는 TNM 병기, PS scale과는 무관한 예후추정 인자로서 임상적 이용아 가능하다고 생각된다.

• 대한세포병리학회지
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• 제6권1호
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• pp.10-17
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• 1995

Anaplastic carcinoma of the thyroid gland is one of the most malignant tumors. Recently, DNA ploidy measured by flow cytometry and image analysis has been suggested as an additional useful indicator of tumor behavior. Studies on the occurrence and clinical significance of DNA aneuploidy in anaplastic carcinoma of the thyroid are rare. In this study, the pattern of DNA ploidy was measured by image analysis on Papanicolaou stained slides in four cases of anaplastic carcinoma and also measured by flow cytometry using paraffin blocks in two cases. In all cases of anaplastic carcinoma, DNA aneuploidy was found by image analaysis. By flow cytometry, one case had a diploid peak and the other case had an aneuploid peak. According to the above results, we conclude that anaplastic carcinoma of the thyroid glands have a high incidence of DNA aneuploidy and image analysis using Papanicolaou stained slides is a useful method in detecting DNA aneuploidy.

• 대한세포병리학회:학술대회논문집
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• 대한세포병리학회 1992년도 제5차 추계학술대회 초록집
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• pp.6.1-6.1
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• 1992

• 한국수산과학회지
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• 제49권5호
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• pp.671-674
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• 2016

We cytogenetically analyzed a triploid King-Nupchi strain of the olive flounder Paralichthys olivaceus to define the simplest, most rapid, and most effective method of ploidy analysis in aquaculture farms. Female triploidy of the flounder King-Nupchi strain was induced by cold shock (3 min post-fertilization at 2-4℃ for 45 min). Triploid induction was confirmed by erythrocyte measurement (nuclear volume, 29.15±2.10 μm³); flow cytometry (2.14±0.03 pg/cell); chromosome count (3N=72); Ag-NOR banding; and silver staining. Silver staining of finned cells obtained using a solid tissue technique was the most effective method of ploidy verification.

• Fisheries and Aquatic Sciences
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• 제20권11호
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• pp.29.1-29.7
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• 2017

The objective of this study was to determine a simple and reliable ploidy identification protocol for the rainbow trout (RT), Oncorhynchus mykiss, in the field condition. To evaluate the ploidy level and compare different detection protocols, triploid RT and gynogenesis were induced by UV irradiation and/or heat shock. The hatching rate at day 30 was 85.2% and the survival rate at day 90 was 69.4% (fingerling). The sex ratio of female RT was 93.75% in the gynogenesis group, illustrating that the UV irradiation inactivated the sperm DNA. The hatching rate and survival rate were 82.0 and 74.7%, respectively, in the triploid-induced group. The triploid induction rate by heat shock procedure was 73.9%. Cytogenetic protocols for ploidy identification such as chromosome counting, erythrocyte nuclear size comparison, and analysis of nucleolar organizing regions (NORs) by silver staining were compared. Silver nitrate staining showed the greatest success rate (22/23 and 32/32 for the triploid-induced group and gynogenesis group, respectively), followed by erythrocyte nuclear size comparison (16/23 and 19/32 for the triploid-induced group and gynogenesis group, respectively) and, lastly, chromosome preparation (2/23 and 6/32 for the triploid-induced group and gynogenesis group, respectively) with the lowest success rate. Based on our findings, silver staining for RT ploidy identification is speculated to be highly applicable in a wide range of research conditions, due to its cost-effectiveness and simplicity compared to other numerous ploidy detection protocols.

• 한국동물생명공학회지
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• 제35권3호
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• pp.223-231
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• 2020

Karyotype analysis is a major work in the process of triploid abalone production for the purpose of productivity and quality improvement. However, the metaphase spreads for karyotype analysis have been prepared just from the larvae at trochophore stage, which has restricted the spectrum of sample correction inhibiting more efficient analysis. Here, we investigated the feasibility of preparing metaphase spreads from the larvae at veliger stage that is the next developmental stage of trochophore. For this, diploid and triploid larvae at trochophore and veliger stages from Pacific abalone (Haliotis discus hannai) were subjected to metaphase spread preparation and its efficiencies were measured and compared each other. As the results, although the efficiencies of metaphase spread preparation were significantly lower in the larvae at veliger stage compared to the ones at trochophore stage regardless of ploidy status, we found that the preparation of metaphase spreads, which showed the clear chromosomal images containing the normal number of chromosomes, was possible from the veliger stage larvae. On the other hands, all larvae used in this study regardless of developmental stage and ploidy did not show colchicine sensitivity. Moreover, no significant difference was observed in cell cycle distribution of the cells comprising larvae between two developmental stages regardless of ploidy status. These suggested that the details of protocol to prepare metaphase spreads from abalone larvae should be optimized depending on its developmental stages. Taken together, we demonstrated the feasibility of preparing metaphase spreads from H. discus hannai veliger stage larvae for karyotype analysis.

• Molecules and Cells
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• 제25권2호
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• pp.163-171
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• 2008

The genus Cynodon comprises ten species. The objective of this study was to evaluate the genetic diversity of Korean bermudagrasses at the morphological, cytological and molecular levels. Morphological parameters, the nuclear DNA content and ploidy levels were observed in 43 bermudagrass ecotypes. AFLP markers were evaluated to define the genetic diversity, and chromosome counts were made to confirm the inferred cytotypes. Nuclear DNA contents were in the ranges 1.42-1.56, 1.94-2.19, 2.54, and 2.77-2.85 pg/2C for the triploid, tetraploid, pentaploid, and hexaploid accessions, respectively. The inferred cytotypes were triploid (2n = 3x = 27), tetraploid (2n = 4x = 36), pentaploid (2n = 5x = 45), and hexaploid (2n = 6x = 54), but the majority of the collections were tetraploid (81%). Mitotic chromosome counts verified the corresponding ploidy levels. The fast growing fine-textured ecotypes had lower ploidy levels, while the pentaploids and hexaploids were coarse types. The genetic similarity ranged from 0.42 to 0.94 with an average of 0.64. UPGMA cluster analysis and principle coordinate analysis separated the ecotypes into 6 distinct groups. The genetic similarity suggests natural hybridization between the different cytotypes, which could be useful resources for future breeding and genetic studies.

• Tuberculosis and Respiratory Diseases
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• 제41권4호
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• pp.354-362
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• 1994

배경 및 방법 : 폐암의 진단 방법에 있어서 객담, 흉수, 기관지 세척액 등에서의 DNA의 aneuploid나 고증식력등의 소견은 폐암의 진단에 보조적으로 의의가 있는 것으로 보고되고 있다. 저자들은 기관지 내시경 검사 중에 조직 생검, 솔질표본의 세포학적 검사와 더불어 유세포계산법을 이용한 솔질 표본의 DNA 배수성 검사를 함께 시행하여, 폐암의 진단율을 높일 수 있는지의 여부를 검토하고자 하였다. 결과: 1) 대상환자 76예중 폐암으로 확진되었던 55예에서는 diploid 37예, aneuploidy 18예(32.7%)이었으나, 양성 질환으로 확인된 21예에서는 모두 diploid 이었고, 세포주기 분석이 기능하였던 48예중 폐암은 35예 이었고 이들중 42.9%(15/35)에서 고증식력을 보였으나, 양성질환 13예에서는 고증식력을 보인 경우가 없었다(p<0.05). 2) DNA분석 소견(aneuploidy나 고증식력을 양성으로 하였을 때)과 세포진검사와의 일치율은 전체 75예중 56예로 74.7% 였다. 3) 폐암 환자에서 세포진검사 민감도는 41.8%(23/55)이었는데, 세포진검사 음성이지만 DNA 검사에서 양성(aneuploidy 혹은 high proliferative activity)을 보인 경우를 부가하였을 때, 민감도는 56.4%(31/55)로 증가하였고(p<0.05), 음성예측도는 38.2%, 특이도는 100% 였다. 4) 1예에서는 기관지내시경을 이용한 조직, 세포진검사, 경피적 폐침흡인 등으로는 진단을 내릴수 없었으나 솔질표본을 이용한 DNA ploidy검사에서 aneuploid로 나타났고, 후에 수술로써 편평상피폐암으로 확진되었다. 5) 폐암 환자중 세포 형태에 따른 DNA Ploidy와 증식력에는 유의한 차이가 없었다. 결론 : 기관지 솔질 표본에서 aneuploid나 고증식력 소견이 폐암을 시사하는 것으로 해석하였을 때, 세포진검사에 부가하여 DNA 배수성 측정과 세포주기 분석을 함께 함으로써 폐암 진단의 예민도를 높일수 있었고, 비교적 특이도가 높은 것으로 사료되었으며, 특히 조직을 얻기 어려운 경우들에서 DNA 분석의 진단적 의의에 대한 계속적인 연구가 필요할 것으로 사료되었다.