No abstract available.
Animals ; Metaphase* ; Mice* ; Oocytes*

The task for chromosome karyotyping and diagnosis is requiring repetitive, time consuming job and high cost even it is done by well-experienced cytogenetists. Therefore an web based chromosome karyotyping instruction system has been established to be able to analyze chromosomes and obtain necessary advises from the database instead of human experts and the database is including 2 divisions with database and agent.For the first of all, database model was constructed with relational database consisting of Patient_DB, image_DB, Disease_DB and Manage_DB. As the second procedure, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes. For the last, independent agent with the inference by knowledge base could enter the inference data into the database.Experimental data were composed of normal chromosomes of 2,736 patients' cases and abnormal chromosomes of 259 patients' cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples.The completed system provides variously morphological information by analysis of normal or abnormal chromosomes and it also makes users enable to control and search the information in a short period with learning of high amount of knowledge.
Amniotic Fluid ; Diagnosis ; Female ; Humans ; Karyotyping* ; Knowledge Bases ; Learning ; Metaphase

Chromosome analysis has been widely used in clinics including prenatal diagnosis. To obtain high-quality metaphase chromosomes, researchers have attempted to modify the methods for chromosome culture, preparation and analysis. Some large research centers also tried to establish standards for quality control. In this paper, modification of methods for the preparation of chromosomes in the last decade is reviewed.
Cells, Cultured ; Chromosomes, Human ; Cytogenetic Analysis ; Humans ; Karyotyping ; Metaphase

Oocyte cryopreservation is widely used for clinical and social reasons. Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures, but not storage time, can alter the gene expression profiles of frozen oocytes. Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase II oocytes remain unknown. Four women (30-32 years old) who had undergone IVF treatment were recruited for this study. RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes (3, 3, 4, and 3 oocytes were cryostored for 1,2, 3, and 12 months) were analyzed at a single-cell resolution. A total of 1987 genes were differentially expressed in the 13 vitrified-thawed oocytes. However, no differentially expressed genes were found between any two groups among the 1-, 2-, 3-, and 12-month storage groups. Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development. Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself, suggesting that long-term cryostorage of human oocytes is safe.
Adult ; Cryopreservation ; Female ; Humans ; Metaphase ; Oocytes ; RNA-Seq ; Vitrification

Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Zl, DXZI, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.
Blastomeres ; Chromosome Aberrations ; DNA Probes ; Fluorescence* ; Germ Cells ; Humans ; In Situ Hybridization* ; Interphase ; Lymphocytes ; Metaphase ; Spermatozoa

PURPOSE: Radiation adaptive response in human peripheral lymphocytes and mouse bone marrow cells was investigated using both metaphase analysis and micronucleus assay. We assessed the correlation between both tests. MATERIALS AND METHODS: Two groups of the human peripheral lymphocytes and mouse bone marrow cells were exposed to low dose (conditioning dose, 0.18 Gy) or high dose (challenging dose, 2 Gy) gamma-rays. The other 4 groups were exposed to low dose followed by high dose after several time intervals (4, 7, 12, and 24 hours, respectively). The frequencies of chromosomal aberrations in metphase analysis and micronuclei in micronucleus assay were counted. RESULTS: Chromosomal aberrations and micronuclei of preexposed group were lower than those of the group only exposed to high dose radiation. Maximal reduction in frequencies of chromosomal aberrations were observed in the group to which challenging dose was given at 7 hour after a conditioning dose (p<0.001). Metaphase analysis and micronucleus assay revealed very good correlation in both human lymphocytes and mouse bone marrow cells (r=0.98, p<0.001; r=0.99, p=0.001, respectively). CONCLUSION: Radiation adaptive response could be induced by low dose irradiation in both human lymphocytes and mouse bone marrow cells. There was a significant correlation between metaphase analysis and micronucleus assay
Animals ; Bone Marrow Cells* ; Bone Marrow* ; Chromosome Aberrations ; Cytogenetics* ; Humans* ; Lymphocytes* ; Metaphase* ; Mice* ; Micronucleus Tests

PURPOSE: Radioiodine (I-131) therapy is an effective modality to reduce both recurrence and mortality rates in differentiated thyroid cancer. Whether higher doses shows higher therapeutic responses was still debatable. The purpose of this study was to validate curve-fitting (CF) method measuring maximum permissible dose (MPD) by a biological dosimetry using metaphase analysis of peripheral blood lymphocytes. MATERIALS AND METHODS: Therapeutic effects of MPD was evaluated in 58 patients (49 females and 9 males, mean age 50+/-11 years) of papillary thyroid cancer. Among them 43 patients were treated with < or =7.4 GBq, while 15 patients with > or =9.25 GBq. The former was defined as low-dose group, and the latter high-dose group. Therapeutic response was defined as complete response when complete disappearance of lesions on follow-up I-131 scan and undetectable serum thyroglobulin levels were found. Statistical comparison between groups were done using chi-square test. P value less than 0.05 was regarded as statistically significant. RESULTS: MPD measured by CF method using tracer and therapeutic doses were 13.3+/-1.9 and 13.8+/-2.1 GBq, respectively (p=0.20). They showed a significant correlation (r=0.8, p< 0.0001). Exposed doses to blood measured by CF and biological methods were 1.54+/-0.03 and 1.78+/-0.03 Gy (p=0.01). They also showed a significant correlation (r=0.86, p=0.01). High-dose group showed a significantly higher rate of complete response (12/15, 80%) as compared to the low-dose group (22/43, 51.2%) (p=0.05). While occurrence of side effects was not different between two groups (40% vs. 30.2%, p=0.46). CONCLUSION: Measurement of MPD using CF method is reliable, and the high-dose I-131 therapy using MPD gains significantly higher therapeutic effects as compared with low-dose therapy.
Female ; Follow-Up Studies ; Humans ; Lymphocytes ; Male ; Metaphase ; Mortality ; Recurrence ; Thyroglobulin ; Thyroid Gland* ; Thyroid Neoplasms*

BACKGROUND: The KCl hypotonic treatment is important for swelling the cells and adequate spreading of chromosomes on the slide. Cytogenetic laboratory usually use 0.075M KCl solution. Sometimes, it is difficult to obtain enough and good quality of metaphase cells, because of inadequate hypotonic treatment. The purpose of this study was to evaluate the mitotic index and band resolution according to the different KCl concentration. METHODS: The group I included blood specimens obtained from 14 newborns (median age 1 day, range 1-8 days) and 4 cord blood. The group II included 16 persons whose median age was 28 years (1-37 years). The blood was cultured in RPMI 1640 medium with fetal calf serum and phytohemagglutinin for 72 hours. Mitosis was arrested by adding colcemid (100 ng/mL). The hypotonic treatment was done by adding different KCl concentration such as 0.075M, 0.068M and 0.057M for 30 minutes at 37 degrees C. The mitotic index was calculated as the number of metaphase cells per total 1,000 cells. The band resolution was evaluated by 2 persons independently. RESULTS: For group I, the mitotic index was not different according to the KCl concentration; 0.075M, 18.8 (5.5~31.5); 0.068M, 22.3 (11~32.5); 0.057M, 20.5 (2.5~29), (P=0.137). The proportion of cells with 400 or more band resolution was significantly higher in specimens treated with 0.068M KCl than those treated with 0.075M KCl; 0.075M, 67.8% (56~92.5); 0.068M, 73.6% (46.1~84.6); 0.057M, 71.6% (63~89.2), (P=0.027). For group II, the results were similar to those of group I. The mitotic index was as follows; 0.075M, 22.3 (5~28); 0.068M, 26 (4~34.5); 0.057M, 21.5 (2.5~36.5), (P=0.568). The proportion of cells with 400 or more band resolution was as follows; 0.075M, 66.6% (42.8~83.3); 0.068M, 69.7% (54.3~87.5); 0.057M, 68.2% (50~78.6) (P=0.04). CONCLUSIONS: For 0.068M or 0.057M KCl treatment, band resolution was improved, while the mitotic index was similar to that of 0.075M KCl. We suggest use of 0.068M or 0.057M KCl hypotonic treatment in addition to 0.075M KCl for chromosome preparation of peripheral blood.
Cytogenetics ; Demecolcine ; Fetal Blood ; Humans ; Infant, Newborn ; Metaphase ; Mitosis ; Mitotic Index*

BACKGROUND: One of major challenge of cytogenetics is to obtain qualitative metaphases to achieve a meaningful analysis. Methotrexate (MTX) synchronization and the Giant Cell Tumor-Conditioned Medium (GCT-CM) have been used to improve the metaphase preparation from hematopoietic malignancies. The purpose of this study is to determine Mitotic Index (MI) of bone marrow samples under several culture conditions that may improve the quality of chromosome preparation. METHODS: Sixty nine bone marrow samples were cultured into 3 groups by traditional methods. The first group was tested for the effect of cell concentration on MI. The second, for the effect of MTX concentration on MI. The third group was classified into 4 subgroups as follows: 1) MTX only in 24 hour culture 2) MTX and GCT-CM in 24 hour culture 3) 48 hour culture without MTX 4) 48 hour culture and GCT-CM. MI was calculated as the ratio of metaphase to interphase cells in 1000 cells. Quality of metaphase was evaluated by classified the metaphase cell into 3 types. RESULTS: The first and second groups revealed no relationship between cell concentration, amount of MTX and MI, respectively. The third group showed significant differences among four subgroups. The MI increased in subgroups using GCT-CM and in the 48 hour culture, with greatest increase in group using 48 hour culture and GCT-CM simultaneously. CONCLUSIONS: The use of GCT-CM medium and prolonged culture improved the quality of metaphase cells and MI. It is therefore beneficial to use these conditions in cytogenetic studies on bone marrow in hematologic disease.
Bone Marrow* ; Cytogenetics ; Giant Cells* ; Hematologic Diseases ; Hematologic Neoplasms ; Interphase ; Metaphase ; Methotrexate ; Mitotic Index*

PURPOSE: Because of difficulty of obtaining metaphase cells from tumor specimens, there are only a few cytogenetic studies in nasal NK/T-cell lymphomas, and so far no consistent specific chromosomal abnormalities have been described. In this study, we have used degenerate oligonucleotide primed PCR (DOP-PCR) and comparative genomic hybridization (CGH) to deter mine chromosomal alterations from 6 nasal NK/T-cell lymphoma tissues dissected from formalin- fixed paraffin-embedded slide sections. MATERIALS AND METHODS: For the isolation of tumor DNA, four 7-micrometer-thick tissue sections from each sample were dewaxed and rehydrated, and areas of high tumor cell content (more than 60%) were dissected and pooled into a tube. Normal DNA was prepared from the peripheral blood of a healthy volunteer. Tumor DNA was labeled with biotin-16-dUTP by DOP-PCR and normal DNA was labeled with digoxigenin-dUTP using a nick translation kit. In CGH, equal amounts of differently labeled DNA from the tumors and normal reference DNA were hybridized simul taneously to normal metaphase chromosomes. They were visualized by different fluordegrees Chromes, and the signal intensities were quantitated separately as gray levels for each chromosome. The over- and underrepresented DNA segments were determined by computation of image ratios and average ratio profiles. RESULTS: Our results show that gains of DNA copy number were more prevalence than DNA losses. The most commonly observed gains were mapped to chromosomal regions of 1p32.2 ter,19 and 20 in 4 of 6 cases (67%). The other frequent gains were found on chromosomes 12q in 3 of 6 cases. The most frequent loss was detected on 6q in 4 of 6 cases(67%), and less fre quently observed on 13q21.1 q34 and 13q14 q34. CONCLUSION: These genomic changes found in specific chromosomal regions are likely to harbor genes of importance in nasal NK/T-cell lymphomagenesis, therefore such cytogenetic mapping of genomic imbalance may be of value for further molecular delineation of NK/T-cell lymphoma.
Chromosome Aberrations ; Comparative Genomic Hybridization* ; Cytogenetics ; DNA ; Healthy Volunteers ; Lymphoma* ; Metaphase ; Polymerase Chain Reaction ; Prevalence