• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.335-340
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• 1997

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 (D18Z1, DXZ1, 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.

• The Korean Journal of Cytopathology
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• v.9 no.2
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• pp.147-153
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• 1998

We have investigated the cervicovaginal smears in order to define the cytologic features of uterine adenocarcinomas. Total 22 cases were reviewed(12 cases from the Sanggye Paik hospital, Inje university and 10 cases from the Korea Cancer Center Hospital) from January 1992 to December 1997. Five cases were endometrial adenocarcinomas and the remaining 17 cases were cervical adenocarcinomas. Seventeen cases of cervical adenocarcinomas were divided into endometrioid(6 cases), endocervical(7 cases), mixed endometrioid and endocervical(1 case), papillary (2 cases), and adenosquamous(1 case) carcinomas. The background of endocervical adenocarcinoma was hemorrhagic or inflammatory. The tumor diathesis was less prominent than that of the squamous cell carcinoma. The prominent features of the endocervical type adenocarcinomas were large and loose clusters, large intracytoplasmic vacuoles, and prominent overlapping and peripheral palisading of nuclei. In contrast, the endometrioid adenocarcinomas showed small and compact clusters, and small intracytoplasmic vacuoles. The detection rate of endometrial adenocarcinoma was lower than that of the endocervical adenocarcinoma.

• Journal of Oral Medicine and Pain
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• v.9 no.1
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• pp.127-138
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• 1984

The author had tried to identify the sex from single tooth by detecting F-body of Y-chromosome in the nucleus of the dental pulp cells of 70 persons aged from 4 to 61 years under a fluorescent miscroscope. The results were as follows : 1. In the cell nuclei of male and female dental pulp at refrigeratory, the rate of F-body appearancd ranged 42-86%(average 61.06%) in male, while it was 0-6%(average 1.86%) in female, indicationg that male could be distinctly differentiated from female by F-body. 2. With male and female dental pulp puterfide by leaving in at room temperature, the rate of F-body appearance ranged 35-58%(average 48.20%) in male, 1-3%(average1.70%) in female, indicating that it was possible to distinguish male and female by F-body. 3. Even in heat-treated male teeth at $100^{\circ}C$,10 mins, the rate of F-body appearace proved to be 32-56%(averaged 42.50%), also indicating the possibility of identifying male. 4. When detecting of F-body in process of time, the rate of F-body appearance did not show major charges. 5. It was reaffirmed that F-body detection method was a positive determination method of male.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.327-333
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• 1994

The purpose of this study was to establish a rapid diagnostic method detecting Aujeszky's disease virus (ADV) DNA in the cultured cell monolayers (PK-15) and tissue sections of ADV(NYJ-1-87)-infected rats and pigs by in situ hybridization(ISH). Detection of specific ADV-DNA in infected cells was conducted by radiolabeled ISH method using $^{32}P-labeled $ DNA probe (BamH1 7 fragment) which contains a 6.3 Kb ADV-DNA insert. Where ADV-DNA was detected by radiolabeled ISH, the deposition of black photographic grains occurred in the nuclei and the cytoplasms of ADV-infected cells. Positive hybridization signal was often observed in the spinal trigerminal nucleus of the pons, the nucleus of the trigerminal ganglion neuron and the epithelial cells of tonsillar crypts. The results suggested that ISH is considered as a highly sensitive and reliable tool for confirmative diagnosis of this viral disease.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.12
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• pp.1681-1690
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• 2012

The aim of this study was to investigate the impact of a reported p53 inhibitor, pifithrin-${\alpha}$ (PFT-${\alpha}$), on preimplantation porcine in vitro fertilized (IVF) embryo development in culture. Treatment of PFT-${\alpha}$ was administered at both early (0 to 48 hpi), and later stages (48 to 168 hpi) of preimplantation development, and its impact upon the expression of five genes related to apoptosis (p53, bak, bcl-xL, p66Shc and caspase3), was assessed in resulting d 7 blastocysts, using real-time quantitative PCR. Total cell numbers, along with the number of apoptotic nuclei, as detected by the in situ cell death detection assay, were also calculated on d 7 in treated and non-treated control embryos. The results indicate that PFT-${\alpha}$, when administered at both early and later stages of porcine IVF embryo development, increases the incidence of apoptosis in resulting blastocysts. When administered at early cleavage stages, PFT-${\alpha}$ treatment was shown to reduce the developmental competence of porcine IVF embryos, as well as reducing the quality of resulting blastocysts in terms of overall cell numbers. In contrast, at later stages, PFT-${\alpha}$ administration resulted in marginally increased blastocyst development rates amongst treated embryos, but did not affect cell numbers. However, PFT-${\alpha}$ treatment induced apoptosis and apoptotic related gene expression, in all treated embryos, irrespective of the timing of treatment. Our results indicate that PFT-${\alpha}$ may severely compromise the developmental potential of porcine IVF embryos, and is a potent apoptotic agent when placed into porcine embryo culture media. Thus, caution should be exercised when using PFT-${\alpha}$ as a specific inhibitor of p53 mediated apoptosis, in the context of porcine IVF embryo culture systems.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1185-1198
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• 2018

The nucleosomal organization of chromatin using histone proteins is a fundamental and ubiquitous feature of eukaryotic nuclei, with the major exception of dinoflagellates. Although a number of recent genomic and transcriptomic analyses have detected numerous histone genes in dinoflagellates, little is known about their expression. Here in, we aimed to investigate the expression pattern of histone genes under nutritional stress, and an attempt was made to detect histone expression at the protein level in Alexandrium pacificum. The presence of histones at the mRNA level was confirmed in this study by the amplification, cloning, and sequencing of 10 different genes. Relative expression profiling of these genes under different growth conditions was determined with real-time PCR and revealed considerable levels of histone transcription in nutritionally stressed cells. We were unable to detect the expression of histones at the protein level even after immunodetection and analysis using mass spectrometry, although a histone-like protein was detected as a major nuclear component. A. pacificum expresses multiple variants of histone, and protein sequences revealed both conservation and divergence with respect to other eukaryotes. We concluded that A. pacificum maintained an active transcription of histone genes within the cell, and enhanced expression of histone genes in nutritional stress strongly suggest that histones have functional significance in dinoflagellates, although expression at the protein level was below our current detection limits, which suggests a limited role of histones in DNA packaging. Finally, the plausible regulation of histone expression at the gene and protein levels in A. pacificum is discussed.

• The Korean Journal of Cytopathology
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• v.14 no.2
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• pp.86-90
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• 2003

Kimura's disease is a chronic inflammatory disorder of unknown cause and is most prevalent among Asians. The cytologic findings of Kimura's disease are significant numbers of eosinophils in a background of lymphoid cells, occasional fragments of collagenous tissue, proliferation oi vessels, and Warthin-Finkeldey polykaryocytes. Among these features, the most important cytologic feature of Kimura's disease is a significant numbers of eosinophils. We experienced a case of Kimura's disease in the parotid gland which we fatted to recognize on cytology due to the apparent paucity of eosinophils. On careful retrograde reviewing of the cytologic findings, a few scattered leukocytes, previously interpreted as polymorphous leukocytes, had bilobed nuclei and coarse green but granular cytoplasm on Papanicolaou preparation. These leukocytes showed obvious orange-red intracyloplasmic granules as in eosionophils on Giemsa stain. The paucity of eosinophils may be due to the thick fibrosis around lymphoid follicles or any technical error during aspiration. Whereas the Warthin-Finkeldey type giant cell is not a sensitive cytologic marker of Kimura's disease, it may be a helpful cytologic feature. To reach a correct cytologic diagnosis of Kimura's disease, It is important to keep in mind that searching for Warthin-Finkeldey type giant cells and evaluation of Giemsa stain for detection of eosinophils would be helpful.

• Development and Reproduction
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• v.10 no.1
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• pp.1-7
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• 2006

Ovarian follicular atresia in mammals is finely regulated by gonadotropins and sex steroid hormones. It is well known that granulosa cell pyknosis is a common cytological feature of atretic follicles in the ovary. The present study hypothesized that granulosa cell pyknosis during follicular atresia might be related to apoptotic process and associated with caspase-3 activation. Healthy (normal) and atretic follicles were isolated from porcine ovaries based on macro-morphological criteria. Isolated follicles were either processed for histological observation or used for collection of granulosa cells by aspiration. Hoechst 33258 staining of the cells showed a significantly higher number of fragmented nuclei, a typical morphological feature of apoptotic cell, in granulosa cells from atretic follicles than those from healthy follicles. In addition, the rate of cell death was significantly higher in granulosa cells from atretic follicles than healthy follicles, as measured by flow-cytometric cell cycle analysis. In situ detection of apoptotic cells by TUNEL revealed that apoptosis was mostly restricted to granulosa cells in follicles. Theca cells were TUNEL-negative. Finally, it has been shown by caspase-3 activity assay that granulosa cells from atretic follicles retain a higher caspase-3 activity compared to healthy follicles. Taken together, it is suggested that granulosa cell degeneration during folliclar atresia occurs by caspase-3-dependent apoptotic fashion.