• The Korean Journal of Mycology
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• v.22 no.1
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• pp.107-116
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• 1994

Interorder somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus in the order Agaricales and Elfvingia applanata in the order Aphyllophorales. The fusants were classified into stable heterokaryons and spontaneously segregated heterokaryons. Hyphae of all fusion products except two strains did not form clamp connections. Out of them, two clamped and three clampless fusants produced mature fruiting bodies by light-dark cycle on sawdust rice bran medium. All of these basidiocarps had clamp connections. Three fusants were analysed with the distribution of progenies and segregation of genetic characters by random spore analyses. The genetic markers were shown to segregate and recombine in the first generation of monospores isolated from basidiocarps. Phenotypes of a large number of auxotrophic progenies were not detected in the two clamped fusants. The aberration ratio of segregants indicated the gene interaction resulting from different genome structure between distantly related species. The polymerase chain reaction (PCR) was adopted for the detection of somatic hybrids nuclear DNA. Four fusants showed a positive results in three kinds of primers. The prominent reaction products are represented by new bands in primer # 87 and # 125. Out of four fusants, two somatic hybrids had non-parental mtDNA patterns when digested with EcoR1 and HindIII. Comparison of somatic hybrids, tissue culture isolates(TC) and multispore germination isolates(MS) were made using esterase isozyme analysis. It is apparent that somatic hybrids had a minor banding patterns which are quite different from those of parents.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.358-367
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• 2021

The World Health Organization (WHO) has declared the coronavirus disease 2019 (COVID-19) as an international health emergency. Current diagnostic tests are based on the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method, which is the gold standard test that involves the amplification of viral RNA. However, the RT-qPCR assay has limitations in terms of sensitivity and quantification. In this study, we tested both qPCR and droplet digital PCR (ddPCR) to detect low amounts of viral RNA. The cycle threshold (CT) of the viral RNA by RT-PCR significantly varied according to the sequences of the primer and probe sets with in vitro transcript (IVT) RNA or viral RNA as templates, whereas the copy number of the viral RNA by ddPCR was effectively quantified with IVT RNA, cultured viral RNA, and RNA from clinical samples. Furthermore, the clinical samples were assayed via both methods, and the sensitivity of the ddPCR was determined to be equal to or more than that of the RT-qPCR. However, the ddPCR assay is more suitable for determining the copy number of reference materials. These findings suggest that the qPCR assay with the ddPCR defined reference materials could be used as a highly sensitive and compatible diagnostic method for viral RNA detection.

• Biomolecules & Therapeutics
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• v.29 no.5
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• pp.551-561
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• 2021

Thyroid cancer is the most common endocrine malignancy. Patients with well-differentiated thyroid cancers, such as papillary and follicular cancers, have a favorable prognosis. However, poorly differentiated thyroid cancers, such as medullary, squamous and anaplastic advanced thyroid cancers, are very aggressive and insensitive to radioiodine treatment. Thus, novel therapies that attenuate metastasis are urgently needed. We found that both PDGFC and PDGFRA are predominantly expressed in thyroid cancers and that the survival rate is significantly lower in patients with high PDGFRA expression. This finding indicates the important role of PDGF/PDGFR signaling in thyroid cancer development. Next, we established a SW579 squamous thyroid cancer cell line with 95.6% PDGFRA gene insertion and deletions (indels) through CRISPR/Cas9. Protein and invasion analysis showed a dramatic loss in EMT marker expression and metastatic ability. Furthermore, xenograft tumors derived from PDGFRA geneedited SW579 cells exhibited a minor decrease in tumor growth. However, distant lung metastasis was completely abolished upon PDGFRA gene editing, implying that PDGFRA could be an effective target to inhibit distant metastasis in advanced thyroid cancers. To translate this finding to the clinic, we used the most relevant multikinase inhibitor, imatinib, to inhibit PDGFRA signaling. The results showed that imatinib significantly suppressed cell growth, induced cell cycle arrest and cell death in SW579 cells. Our developed noninvasive apoptosis detection sensor (NIADS) indicated that imatinib induced cell apoptosis through caspase-3 activation. In conclusion, we believe that developing a specific and selective targeted therapy for PDGFRA would effectively suppress PDGFRA-mediated cancer aggressiveness in advanced thyroid cancers.

• Korean Journal of Animal Reproduction
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• v.9 no.2
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• pp.140-147
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• 1985

Seventy four Holstein heifers were randomly assigned over three trials to PRID-7+PG-6 and Synchromate B-9 regimens to synchronize estrus cycle for embryo transfer. Sexual behaviors; moounting, standing, orientation, chin-resting, sniffing, licking, rubbing and butting, vaginal swelling and mucus discharge were observed between 06-08, 12-14 and 18-20 h on 1st day and 00-02, 06-08, 12-14 and 18-20 h on 2nd day after removal of hormones. Synchromate-B treatment (81.6%) showed higher synchronized estrus rate than PRID treatment (77.8%) during observation period. Standing estrus was observed within 74 h after PG injection in PRID and within 52 h after removal of implant in Synchromate-B. About 68% of heifers in PRID and 74% of heifers in Synchromate-B showed standing estrus between 0-14 h on 2nd day after removal of the hormones. Synchromate-B resulted in a tighter synchrony of standing estrus than PRID. Incidence of average mounting and standing per head during observation period was 22.3 and 16.6 in PRID and 28.1 and 13.6 in Synchromate-B. The PRID showed peak in active mounting at 18-20 h on 1st day, however, the Synchromate-B showed at 0-2 h on 2nd day after removal of hormone. Active standing was shown between 18 h on 1st day to 20h on 2nd day in PRID, however, between 0-14 h on 2nd day after removal of hormone is Synchromate-B. There was slight difference in pattern of active mounting and standing during estrus between PRID and Synchromate-B. Conception rate of synchronized heifers transferred with fresh and frozen embryos by non-surgical and surgical methods was higher in synchromate-B (62.5%) than in PRID (38.5%). Chin-resting showed highest incidence among 6 sexual behavioral components in the both treatments. Synchromate-B showed higher incidence of chin-resting (16.6) than PRID (10.7). Synchromate-B group showed also higher incidence of orientation, sniffing and butting than PRID group. Synchromate-B resulted in more active sexual behaviors than PRID. The pattern of incidence of chin-resting, licking and butting was almost symmetrical in PRID with their peak values at 6-8 h on 2nd day, however in Synchromate-B chin-resting and sniffing was symmetrical with their peak values at 12-14 h on 2nd day after removal of hormone. There was tendency to increase vaginal swelling according to time passage of synchronized estrus in the both treatments. Incidence of mucus discharge in Synchromate-B was slightly higher than in PRID. Twenty to 40% was false negative in conception rate by tall painting before re-estrus day as judged by rectal palpation.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.22 no.2
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• pp.164-173
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• 2000

The p16 protein is a cyclin dependent kinase inhibitor that inhibits cell cycle progression from $G_1$ phase to S phase in cell cycle. Many p16 gene mutations have been noted in many cancer-cell lines and in some primary cancers, and alterations of p16 gene function by DNA methylation have been noticed in various kinds of cancer tissues and cell-lines. There have been a large body of literature has accumulated indicating that abnormal patterns of DNA methylation (both hypomethylation and hypermethylation) occur in a wide variety of human neoplasma and that these aberrations of DNA methylation may play an important epigenetic role in the development and progression of neoplasia. DNA methylation is a part of the inheritable epigenetic system that influences expression or silencing of genes necessary for normal differentiation and proliferation. Gene activity may be silenced by methylation of up steream regulatory regions. Reactivation is associated with demethylation. Although evidence or a high incidence of p16 alterations in a variety of cell lines and primary tumors has been reported, that has been contested by other investigators. The precise mechanisms by which abnormal methylation might contribute to carcinogenesis are still not fully elucidated, but conceivably could involve the modulation of oncogene and other important regulatory gene expression, in addition to creating areas of genetic instability, thus predisposing to mutational events causing neoplasia. There have been many variable results of studies of head and neck squamous cell carcinoma(HNSCC). This investigation was studied on 13 primary HNSCC for p16 gene status by protein expression in immunohistochemistry, and DNA genetic/epigenetic analyzed to determine the incidence, the mechanisms, and the potential biological significance of its Inactivation. As methylation detection method of p16 gene, the methylation specific PCR(MSP) is sensitive and specific for methylation of any block of CpG sites in a CpG islands using bisulfite-modified DNA. The genomic DNA is modified by treatment with sodium bisulfate, which converts all unmethylated cytosines to uracil(thymidine). The primers designed for MSP were chosen for regions containing frequent cytosines (to distinguish unmodified from modified DNA), and CpG pairs near the 5' end of the primers (to provide maximal discrimination in the PCR between methylated and unmethylated DNA). The two strands of DNA are no longer complementary after bisulfite treatment, primers can be designed for either modified strand. In this study, 13 paraffin embedded block tissues were used, so the fragment of DNA to be amplified was intentionally small, to allow the assessment of methylation pattern in a limited region and to facilitate the application of this technique to samlples. In this 13 primary HNSCC tissues, there was no methylation of p16 promoter gene (detected by MSP and automatic sequencing). The p16 protein-specific immunohistochemical staining was performed on 13 paraffin embedded primary HNSCC tissue samples. Twelve cases among the 13 showed altered expression of p16 proteins (negative expression). In this study, The author suggested that low expression of p16 protein may play an important role in human HNSCC, and this study suggested that many kinds of genetic mechanisms including DNA methylation may play the role in carcinogenesis.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.191-198
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• 2006

The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.

• Korean Journal of Remote Sensing
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• v.36 no.2_2
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• pp.249-261
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• 2020

Coastal monitoring using multiple platforms/sensors is a very important tools for accurately understanding the changes in offshore marine environment and disaster with high temporal and spatial resolutions. However, integrated observation studies using multiple platforms and sensors are insufficient, and none of them have been evaluated for efficiency and limitation of convergence. In this study, we aimed to suggest an integrated observation method with multi-remote sensing platform and sensors, and to diagnose the utility and limitation. Integrated in situ surveys were conducted using Rhodamine WT fluorescent dye to simulate various marine disasters. In September 2019, the distribution and movement of RWT dye patches were detected using satellite (Kompsat-2/3/3A, Landsat-8 OLI, Sentinel-3 OLCI and GOCI), unmanned aircraft (Mavic 2 pro and Inspire 2), and manned aircraft platforms after injecting fluorescent dye into the waters of the South Sea-Yeosu Sea. The initial patch size of the RWT dye was 2,600 ㎡ and spread to 62,000 ㎡ about 138 minutes later. The RWT patches gradually moved southwestward from the point where they were first released,similar to the pattern of tidal current flowing southwest as the tides gradually decreased. Unmanned Aerial Vehicles (UAVs) image showed highest resolution in terms of spatial and time resolution, but the coverage area was the narrowest. In the case of satellite images, the coverage area was wide, but there were some limitations compared to other platforms in terms of operability due to the long cycle of revisiting. For Sentinel-3 OLCI and GOCI, the spectral resolution and signal-to-noise ratio (SNR) were the highest, but small fluorescent dye detection was limited in terms of spatial resolution. In the case of hyperspectral sensor mounted on manned aircraft, the spectral resolution was the highest, but this was also somewhat limited in terms of operability. From this simulation approach, multi-platform integrated observation was able to confirm that time,space and spectral resolution could be significantly improved. In the future, if this study results are linked to coastal numerical models, it will be possible to predict the transport and diffusion of contaminants, and it is expected that it can contribute to improving model accuracy by using them as input and verification data of the numerical models.

• Journal of Embryo Transfer
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• v.27 no.3
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• pp.127-131
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• 2012

This study was performed to investigate the effects of parity and season on the embryo production in superovulated Hanwoo cows. Superovulation was performed from 1 to 8 times by repeated superovulation treatment of Hanwoo cows (n = 22). Irrespective of estrous cycle, donor cows were received a CIDR, progesterone (50 mg) and estradiol benzoate (2.5 mg). After 4.5 days, the donor cows were superovulated with total 28AU FSH (Antorin R-10) administrated twice daily in a decreasing dose for 4 days. On $6^{th}$ and $7^{th}$ of FSH injection, 2.5 mg and 15 mg $PGF_2{\alpha}$ were injected i.m, respectively. CIDR was removed at the $7^{th}$ FSH injection. The donor cows received $200{\mu}g$GnRH at 48 hrs after $1^{st}$ $PGF_2{\alpha}$ injection. The donor cows were artificially inseminated three times after estrous detection at 12 hr intervals and embryos were recovered 7 days after estrous detection. The mean number of total ova, transferrable embryos, degenerated embryos and unfertilized oocytes were $11.6{\pm}7.9$, $5.5{\pm}4.4$, $3.0{\pm}3.3$ and $2.6{\pm}4.1$ per donor cows, respectively. A higher number of total ova were recovered in parity 3~5 ($14.3{\pm}1.3$) than 1~2 ($8.9{\pm}1.9$, P<0.05). The number of recovered normal embryos is significantly higher in parity 3~5 ($7.3{\pm}0.8$) than that of over 6 ($3.7{\pm}1.5$). Significantly higher number of total ova and normal embryos were recovered in summer ($16.4{\pm}2.3$, $8.1{\pm}1.4$) than in autumn ($10.1{\pm}1.8$, $4.5{\pm}1.1$) and winter ($6.3{\pm}1.8$, $3.3{\pm}1.1$), respectively (P<0.05). Transferable embryos were significantly higher in summer ($7.6{\pm}1.3$) than in winter ($3.0{\pm}1.0$, P< 0.05). The results were showed that parity and season affecting on the production of embryos in superovulated Hanwoo.

• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.339-353
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• 1994

Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.

• Proceedings of the Korean Institute of Intelligent Systems Conference
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• 1993.06a
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• pp.1051-1054
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• 1993

The International Atomic Energy Agency's Statute in Article III.A.5 allows it“to establish and administer safeguards designed to ensure that special fissionable and other materials, services, equipment, facilities and information made available by the Agency or at its request or under its supervision or control are not used in such a way as to further any military purpose; and to apply safeguards, at the request of the parties, to any bilateral or multilateral arrangement, or at the request of a State, to any of that State's activities in the field of atomic energy”. Safeguards are essentially a technical means of verifying the fulfilment of political obligations undertaken by States and given a legal force in international agreements relating to the peaceful uses of nuclear energy. The main political objectives are: to assure the international community that States are complying with their non-proliferation and other peaceful undertakings; and to deter (a) the diversion of afeguarded nuclear materials to the production of nuclear explosives or for military purposes and (b) the misuse of safeguarded facilities with the aim of producing unsafeguarded nuclear material. It is clear that no international safeguards system can physically prevent diversion. The IAEA safeguards system is basically a verification measure designed to provide assurance in those cases in which diversion has not occurred. Verification is accomplished by two basic means: material accountancy and containment and surveillance measures. Nuclear material accountancy is the fundamental IAEA safeguards mechanism, while containment and surveillance serve as important complementary measures. Material accountancy refers to a collection of measurements and other determinations which enable the State and the Agency to maintain a current picture of the location and movement of nuclear material into and out of material balance areas, i. e. areas where all material entering or leaving is measurab e. A containment measure is one that is designed by taking advantage of structural characteristics, such as containers, tanks or pipes, etc. To establish the physical integrity of an area or item by preventing the undetected movement of nuclear material or equipment. Such measures involve the application of tamper-indicating or surveillance devices. Surveillance refers to both human and instrumental observation aimed at indicating the movement of nuclear material. The verification process consists of three over-lapping elements: (a) Provision by the State of information such as - design information describing nuclear installations; - accounting reports listing nuclear material inventories, receipts and shipments; - documents amplifying and clarifying reports, as applicable; - notification of international transfers of nuclear material. (b) Collection by the IAEA of information through inspection activities such as - verification of design information - examination of records and repo ts - measurement of nuclear material - examination of containment and surveillance measures - follow-up activities in case of unusual findings. (c) Evaluation of the information provided by the State and of that collected by inspectors to determine the completeness, accuracy and validity of the information provided by the State and to resolve any anomalies and discrepancies. To design an effective verification system, one must identify possible ways and means by which nuclear material could be diverted from peaceful uses, including means to conceal such diversions. These theoretical ways and means, which have become known as diversion strategies, are used as one of the basic inputs for the development of safeguards procedures, equipment and instrumentation. For analysis of implementation strategy purposes, it is assumed that non-compliance cannot be excluded a priori and that consequently there is a low but non-zero probability that a diversion could be attempted in all safeguards ituations. An important element of diversion strategies is the identification of various possible diversion paths; the amount, type and location of nuclear material involved, the physical route and conversion of the material that may take place, rate of removal and concealment methods, as appropriate. With regard to the physical route and conversion of nuclear material the following main categories may be considered: - unreported removal of nuclear material from an installation or during transit - unreported introduction of nuclear material into an installation - unreported transfer of nuclear material from one material balance area to another - unreported production of nuclear material, e. g. enrichment of uranium or production of plutonium - undeclared uses of the material within the installation. With respect to the amount of nuclear material that might be diverted in a given time (the diversion rate), the continuum between the following two limiting cases is cons dered: - one significant quantity or more in a short time, often known as abrupt diversion; and - one significant quantity or more per year, for example, by accumulation of smaller amounts each time to add up to a significant quantity over a period of one year, often called protracted diversion. Concealment methods may include: - restriction of access of inspectors - falsification of records, reports and other material balance areas - replacement of nuclear material, e. g. use of dummy objects - falsification of measurements or of their evaluation - interference with IAEA installed equipment.As a result of diversion and its concealment or other actions, anomalies will occur. All reasonable diversion routes, scenarios/strategies and concealment methods have to be taken into account in designing safeguards implementation strategies so as to provide sufficient opportunities for the IAEA to observe such anomalies. The safeguards approach for each facility will make a different use of these procedures, equipment and instrumentation according to the various diversion strategies which could be applicable to that facility and according to the detection and inspection goals which are applied. Postulated pathways sets of scenarios comprise those elements of diversion strategies which might be carried out at a facility or across a State's fuel cycle with declared or undeclared activities. All such factors, however, contain a degree of fuzziness that need a human judgment to make the ultimate conclusion that all material is being used for peaceful purposes. Safeguards has been traditionally based on verification of declared material and facilities using material accountancy as a fundamental measure. The strength of material accountancy is based on the fact that it allows to detect any diversion independent of the diversion route taken. Material accountancy detects a diversion after it actually happened and thus is powerless to physically prevent it and can only deter by the risk of early detection any contemplation by State authorities to carry out a diversion. Recently the IAEA has been faced with new challenges. To deal with these, various measures are being reconsidered to strengthen the safeguards system such as enhanced assessment of the completeness of the State's initial declaration of nuclear material and installations under its jurisdiction enhanced monitoring and analysis of open information and analysis of open information that may indicate inconsistencies with the State's safeguards obligations. Precise information vital for such enhanced assessments and analyses is normally not available or, if available, difficult and expensive collection of information would be necessary. Above all, realistic appraisal of truth needs sound human judgment.