• Title/Summary/Keyword: mitotic frequency

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Construction of Starch-Fermenting Yeast Using Protoplast Fusion Technique (원형질체 융합 방법을 이용한 전분 발효성 효모의 개발)

  • Chun, Soon-Bai;Bai, Suk;Lee, Kee-Young;Kim, Soo-Hyun;Lee, Jin-Jong;Kim, Sang-Moon
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.120-127
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    • 1990
  • Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was $2.9\times 10^{-3}$ and $8.5\times 10^{-3}$, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was $8.8\times 10^{-6}$ and $9.5\times 10^{-4}$, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that $\alpha$-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.

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Effect of Brown Rice Extract on Mitomycin C-Induced Chromosome Aberration in Cultured CHL Cells (현미 추출물이 Mitomycin C로 유발된 CHL 세포의 염색체 이상에 미치는 영향)

  • Chun, Hyang-Sook;Kim, In-Ho;Kim, Hyun-Jung
    • Korean Journal of Food Science and Technology
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    • v.27 no.6
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    • pp.1003-1007
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    • 1995
  • The effect of brown rice extract on mitomycin C(MMC)-induced chromosome aberration was examined in cultured Chinese hamster lung(CHL) cells, after induction of chromosome aberration and mitotic index in CHL cells cultured with MMC were observed. There were no significant differences between mitotic indices of CHL cells treated with DMSO, and MMC and brown rice extract. The frequency of chromosome aberration showed dose-dependent relationship in CHL cells treated with $0.2{\sim}3.0\;{\mu}g$/assay of MMC. But chromosome aberrations could not be assayed Our to cytotoxicity of MMC when its concentrations were above $3.0\;{\mu}g$/assay. Chromatid type, especially gap and break, of chromosome aberration were most frequently observed. When CHL cells treated with $2.0\;{\mu}g$/assay of MMC and brown rice extracts of concentration ranging $0.75{\sim}10.0\;{\mu}g$/assay were incubated, frequencies of chromosome aberration induced by MMC were significantly decreased at above concentrations(p<0.01, p<0.05). As concentration of brown rice extract was increased, frequencies of chromosome aberration was decreased $7{\sim}30%$, in some irregularity.

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Cotransformation of Saccharomyces cerevisiae with Heterogenous Plasmids (이종(異種) Plasmid에 의한 Saccharomyces cerevisiae의 동시형질(同時形質) 전환(轉換))

  • Kang, Byung Tae;Park, Jong Sung;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.52-58
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    • 1987
  • The yeast S. cerevisiae DBY747 was transformed with E. C - S. C shuttle vector YIp5, YEp13 and YRp7 by the method of spheroplast. The transformation frequency of YEp13 and YRp7 in S. cerevisiae DBY747 was $1.2{\times}10^3$ and $1.0{\times}10^2$ per $10{\mu}g$ of DNA, respectively. The transformants with YIp5 plasmid incapable of autonomous replication in S. cerevisiae were not detected in the condition of this experiment, but YIpS plasmid expressed the gene carried on it when cotransformed with a helper plasmid such as YEp13 or YRp7 : autonomously replicating plasmid. When plasmids were used in covalently closed circular form, cotransformation frequency of Ylp5-YEpl3 and Ylp5-YRp7 was 210 and 95 per $10{\mu}g$ of DNA, respectively. In cotransformation of linear plasmids, transformation frequency of the same cohesive ends was similar to that of noncomplementary cohesive ends. Transformants by the cotransformation with circular plasmids have been shown much higher frequency than with linear plasmids in S. cerevisiae DBY 747. The mitotic segregation stability test suggested that the cotransformant of YIpS-YEp13 was more stable than that of YIpS-YRp7.

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Fine Needle Aspiration Cytology of Hepatocellular Carcinoma - A Study on 247 Cases - (간세포암종의 세침흡인 세포학적 소견 - 247예에 대한 연구 -)

  • Lee, Kwang-Gil;Lee, Jong-Tae;Choi, Soo-Im;Park, Chan-Il
    • The Korean Journal of Cytopathology
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    • v.1 no.1
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    • pp.1-17
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    • 1990
  • Hepatocellular carcinoma (HCC) is malignant tumor frequently occurring in Koreans. There have been few reports regarding the cytologic findings of fine needle aspiration (FNA) of HCC. Most have suggested a diagnostic problem in the cytology distinguishing HCC from some benign hepatic lesions-for example, a regeneration nodule in cirrhosis and liver cell adenoma. In spite of its high frequency in Korea, no cytologic study has been reported, concerning the FNA of HCC. In an attempt to achieve cytologic criteria for the diagnosis of HCC, the authors studied retrospectively cytopathologic findings of 247 cases of HCC. These cases were confirmed either by histoiogic examination including lobectomy, biopsy, or ceil block material, or, when tissue diagnosis was unavailable, by a high serum alpha-fetoprotein level (over 400 I. U.). All aspiration smears were stained by the Papanicolaou method. In each case, the smears were analyzed for cell patterns and various cytomorphology of the tumor cells. The smear background was assessed for the presence of tumor cell necrosis and inflammatory components and compared to that of metastatic carcinomas. The cell patterns were classified as trabecular, acinar, dispersed, and irregular. The cytologic parameters analyzed included the degree of nuclear atypia and the presence of mitoses, intranuclear cytoplasmic inclusions, nucleolar prominency, endothelial lining, multinucleated giant cells, eosinophiic globules, bile, and Mallory body. Most of the FNA of HCC showed markedly cellular smears. The tumor cells were most frequently arranged in a trabecular pattern (80.3%). The irregular (12.6%), the acinar (5.5%), and the dispersed patterns (1.7%) followed in decreasing frequency. Individual hepatoma cells were larger than normal liver cells. However, they had morphologic features characteristic of the hepatic cells the cells were round or polygonal, their cytoplasm was abundant and granular with eosinophilic or amphophilic stainability, and their nuclei were round to oval, located centrally, and tended to have prominent nucleoli. Anaplasia and pleomorphism of tumor cells were generally mild to moderate. These findings existed even in very well differentiated cases. Mitotic figures were present in about 85% of the cases. Prominent nucleoli were observed only in about half the cases. The frequency of other cytologic features was as follows intranuclear cytoplasmic inclusion in 86.8% : endothelial lining in 56.1% : bile in 19.8% : and giant cells in 60.1%. Clear cells were often present in 11.7%, Most aspiration smears of HCC displayed clean background without necrosis or inflammatory material in contrast to the dirty, necrotic background of metastatic cancers and cholangiocarcinomas. Based on the above mentioned features, it is suqqested that the cytologic critieria most important for the diagnosis of HCC include a markedly cellular smear, trabecular pattern, hepatocytoid appearance of tumor cells, endothelial lining, the presence of bile, giant cells, intranuclear cytoplasmic inclusions, and prominent nucleoli, Among these, trabecular pattern, endothelial lining, giant cells and clean smear background are points to be considered in differentiating HCC from metastatic and cholangiocellular carcinoma.

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CULTURED HUMAN ORAL KERATINOCYTES; ULTRASTRUCTURAL STUDY (배양된 사람치은각화상피세포의 미세구조)

  • Kwon, Yong-Dae;Lee, Baek-Soo;Jue, Sung-Sook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.3
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    • pp.231-239
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    • 1999
  • In oral and maxillofacial surgery, there are many cases requiring the graft of epidermal tissues such as maxillectomy, and vestibuloplasty. There have been so many challenges for the culture of the epidermal tissue. Observing the ultrastructure of the cultured human oral kertinocytes, we could compare this findings with that of in vivo ones. With that, we could find the differencies and similarities between cultured cells and in vivo ones, and evaluate the clinical applications of cultured tissue. Human gingiva was obtained and the specimen was explanted on 24-well plate. Two types of culture media were used in this culture system. One was for the growth of the keratinocytes (Media I), and the other was for the stratification (Media II). Media I had special ingredients for the epidermal growth. Those were 0.5% dimethyl sulfoxide (DMSO), 30ng/ml of epidermal growth factor (EGF), 30ng/ml of cholera toxin, and $5{\mu}g/ml$ of transferrin. We cultured the oral keratinocytes for 3 weeks, and at that time the cultured keratinocytes were processed to prepare the specimen for the TEM study. The results were as follows.; 1. In the phase contrast micrograph, epidermal outgrowth firstly appeared on the 3rd day after explantation, and the growing keratinocytes were activley mitotic, and had polygonal shape and increased N/C ratio. 2. In the phase contrast micrograph, the outer most cells exhibited areas where broad cytoplasmic processes extended out onto the culture subtratum(fan-like appaearance). 3. In the TEM micrographs, the cultured keratinocytes showed stratification. The cells were in elongated form, and there were no morphologic differencies among the layers usually found in the in vivo gingiva. 4. Most of cellular organelles underwent lysis, and keratohyaline granules were seen. Tonofibrils were dispersed in the cytoplasm. 5. The cells were interconnected by desmosomes, and their frequency of distribution was considered to be lower than that of in vivo keratinocytes. 6. We could conclude the cultured oral keratinocytes exhibited signs of terminal differentiation.

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Effects of Caffeine on Maturation-Promoting Factor (MPF) Activity in Bovine Oocytes and on the Development of Somatic Cell Nuclear Transfer Embryos in White-Hanwoo

  • Lee, Joon-Hee;Lee, Hee-Gyu;Baik, Sang-Ki;Jin, Sang-Jin;Moon, Song-Yi;Eun, Hye-Ju;Kim, Tae-Suk;Ko, Yeoung-Gyu;Kim, Sung-Woo;Park, Hae-Geum;Park, Soo-Bong
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.261-267
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    • 2012
  • The technique of SCNT is now well established but still remains inefficient. The in vitro development of SCNT embryos is dependent upon numerous factors including the recipient cytoplast and karyoplast. Above all, the metaphase of the second meiotic division (MII) oocytes have typically become the recipient of choice. Generally high level of MPF present in MII oocytes induces the transferred nucleus to enter mitotic division precociously and causes NEBD and PCC, which may be the critical role for nuclear reprogramming. In the present study we investigated the in vitro development and pregnancy of White-Hanwoo SCNT embryos treated with caffeine (a protein kinase phosphatase inhibitor). As results, the treatment of 10 mM caffeine for 6 h significantly increased MPF activity in bovine oocytes but does not affect the developmental competence to the blastocyst stage in bovine SCNT embryos. However, a significant increase in the mean cell number of blastocysts and the frequency of pregnant on 150 days of White-Hanwoo SCNT embryos produced using caffeine treated cytoplasts was observed. These results indicated that the recipient cytoplast treated with caffeine for a short period prior to reconstruction of SCNT embryos is able to increase the frequency of pregnancy in cow.

Protoplast Fusion of Lactose Assimilating Yeasts (유당 자화 효모간의 원형질체 융합)

  • Choi, Won-Ki;Chun, Soon-Bai;Lee, Yong-Kyu;Bai, Suk;Lee, Jin-Jong;Lee, Hyang
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.188-196
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    • 1988
  • Intergeneric or intraspecific protoplast fusion between Saccharomyces cerevisiae X-2180-1A, Candida pseudotropicalis ATCC 8619 and CBS 607 was attempted to produce ethanol from lactose containing materials. Teh intergeneric fusion frequency between Saccharomyces cerevisiae X-2180-1A (ade rho) and Candida pseudotropicalis CBS 607 (his met) was $1.0\times 10^{-5}$. These values exhibited approximately 2-3.5 fold increase when compared with fusion frequency obtained without the treatment of bovine serum albumin, myoinositol and ergosterol, suggesting that these compounds may improve intergeneric of intraspecific protoplast fusion. Nuclear fusion appears to occur in fusants between intergenera(S. cerevisiae+C. pseudotropicalis) and intraspecies (C. pseudotropicalis strains) as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and isolation of recombinants after mitotic segragation. It was also found that alcohol production from intraspecific hybrids was somewhat increased when compared with that from their parents.

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Selenium Effect on the Frequency of SCEs Induced by Heavy Metals in Human Lymphocytes (Selenium이 mercury, cadmium 및 chromium에 의한 자매염색분체교환(姉妹染色分體交換)의 빈도(頻度)에 미치는 영향(影響))

  • Koh, Dai-Ha;Ki, No-Suk
    • Journal of Preventive Medicine and Public Health
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    • v.23 no.1 s.29
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    • pp.1-10
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    • 1990
  • The protective effect of sodium selenite($Na_2SeO_3$) against the cytogenetic toxicity of heavy metals was investigated on human whole-blood cultures in relation to induction of sister chromatid exchange (SCE) in secondary metaphase chromosome. Methylmercury chloride($CH_3HgCl$), cadmium chloride($CdCl_2$), potassium dichromate($K_2Cr_2O_7$), and sodium selenite caused to the typically dose-dependent increase in sister chromatid exchanges (SCEs) by the concentrations ranging from $0.3{\mu}M\;to\;10{mu}M$. However, the inductions of sister chromatid exchanges by methylmercury chloride or cadmium chloride were inhibited by the simultaneous addition of sodium selenite $1.2{mu}M$. The frequencies of SCE were decreased to the level of control in the molar ratios as 2:1, 1:1, 1:2, and 1:4 of selenium selenite vs. methylmercury chloride, and as 1:1 and 1:2 of selenium selenite vs. cadmium chloride, while the frequencies of SCE induced by potassium dichromate were not changed by the addition of sodium selenite in culture condition. Mitotic indices were decreased in the higher concentrations of chemicals and not significantly changed by the simultaneous addition of sodium selenite to the culture condition containing each chemicals.

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Evaluation of Radiation-induced Apoptosis in Seminiferous Tubule of ICR Mouse after Gamma Irradiation. (감마선을 조사한 ICR 마우스 정세관에서 apoptosis 발생 평가)

  • Jang, Jong-Sik;Kim, Joong-Sun;Kim, Jong-Choon;Kim, Sung-Ho
    • Journal of Life Science
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    • v.19 no.6
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    • pp.799-803
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    • 2009
  • The killing of male germ cells by radiation and other toxicants has recently been attributed to apoptosis, but a critical evaluation of the presence of the different features of apoptosis in each epithelial stage has not been performed. In this study, mouse testes exposed to radiation were examined by light microscopy and terminal transferase-mediated end labeling (TUNEL) with periodic acid-Schiff (PAS) stains to determine whether the cells were apoptotic according to several criteria. Apoptosis was easily recognized by the presence of peroxidase-stained, entirely apoptotic bodies. In the TUNEL-positive cells or bodies, the stained products correlated precisely with the typical morphologic characteristics of apoptosis as seen at the light microscopic level. The changes that occurred from 0 to 24 hours after exposing the mice to 2 Gy of gamma-rays (2 Gy/min) were examined. The numbers of apoptotic cells reached a peak at 12 hours after irradiation and then declined. The mice that received 0-8 Gy of gamma-rays were examined 8 hours after irradiation. Dose-response relationships were generated for each stage of the epithelial cycle by counting TUNEL-positive cells. The dose-response curves were linear- quadratic [y=(-0.014${\pm}$0.009)$D^{2}$+(0.31${\pm}$0.697)D+0.3575. Where y=the number of apoptotic cells per seminiferous tubule, and D=the irradiation dose in Gy, $r^{2}$=0.9] and there was a significant relationship between the frequency of apoptotic cells and the radiation dose. Although the maximum response was produced by 8 Gy, even 0.5 Gy induced marked changes. These changes were most pronounced in B spermatogonia of stage V and the spermatocyte at the mitotic cells of stage XII.

Stability of Human Centromeric Alphoid DNA Repeat during Propagation in Recombination-Deficient Yeast Strains (효모의 재조합 변이주를 이용한 인간 Centromeric Alphoid DNA Repeat의 안정성에 관한 연구)

  • Kim, Kwang-Sup;Shin, Young-Sun;Lee, Sang-Yeop;Ahn, Eun-Kyung;Do, Eun-Ju;Park, In-Ho;Leem, Sun-Hee;SunWoo, Yang-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.243-249
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    • 2007
  • The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.