• Title/Summary/Keyword: cytokinesis

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Evaluation of micronucleus frequency in cytokinesis-blockedlymphocytes of cattle in the vicinity of Uljin nuclear power station (세포질 분열 차단 림프구를 이용한 울진원자력발전소 주변 소의 미소핵 발생 평가)

  • Kim, Se-ra;Kang, Chang-mo;Kim, Sung-ho
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.343-348
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    • 2004
  • Cytogenetic and hematological analysis was performed in peripheral blood of cattle in the vicinity of Uljin nuclear power station and control area. The frequency of micronuclei(MN) in peripheral blood lymphocytes from cattle was used as a biomarker of radiobiological effects resulting from exposure to environmental radiation. An estimated dose of radiation was calculated by a best fitting linear-quadratic model based on the radiation-induced MN formation from the bovine lymphocytes exposed in vitro to radiation over the range from 0 Gy to 4 Gy. MN ratio in lymphocytes of cattle from Uljin nuclear power station and control area were 8.90/1,000 and 9.60/1,000, respectively. There were no significant differences in MN frequencies and hematological values in cattle between Uljin and control area.

Sequence analysis of the schizosaccharomycs pombe homologue of the CDC3 gene in saccharomyces cerevisiae

  • Kim, Hyong-Bai
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.350-354
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    • 1995
  • Saccharomyces cervisiae has a highly ordered ring of filaments that lies just inside the cytoplasmic membrane in the region of the mother-bud neck. Mutants defective in any one of the our cell division cycle genes (CDC3, CDC10, CDC11, CDC12) fail to form these filaments and exhibit a pleiotropic phenotype that includes failure to complete cytokinesis and abnormal bud growth. However, the role of the filament is not clear. In order to find out the role of filament, the similar gene in S pombe (called cdc103$\^$+) to the CDC3 was cloned and sequenced. Here I report the sequence analysis of the cdc103$\^$+/ ) to the CDC3 was cloned and sequenced. Here I report the sequence analysis of the cdc103$\^$+/. Comparison of the predicted amino acid sequences of cdc103$\^$+/ and CDC3 revealed that they share significant similarity (43% identity and 56% identity or similarity) to each other.

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The changes in the Amounts of SH Compounds in Chlorella during the Synchronous Culture (Chlorella의 생활사를 통한 -SH화합물 함량의 변화)

  • 최호형;이영녹
    • Korean Journal of Microbiology
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    • v.19 no.1
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    • pp.8-13
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    • 1981
  • The content of sulfhydryl compounds in Chlorella cells during the life cycle in the synchronous culture is determined spectrophotomatically at 250nm(pH7.0) using p-CMB as SH-reagent. The changes in the content of-SHl compounds and protein in Chlorella cells is measured during the life cycle in connection with cell division and analyzed. 1) The amounts of total ninhydrin reactive substance increased with growth of cells but increased the more at the $L_4$ stage(cytokinesis stage) than at the $L_2$ stage (nuclear division stage). 2) The sulfhydryl content of Chlorella cells increased strikingly at the $L_2$ stage and decreased markedly at the $L_4$ stage. 3) The amounts of values -SH/protein showed a peak at the $L_2$ stage. The increase of the amount of total-SH compounds of cells during the nuclear division period was considered to be caused by the weighty roles of protein-SH groups for the formation of nuclear division apparatus and for the enzyme activity.

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Frequency of Micronuclei in Lymphocytes Following Gamma and Fast-neutron Irradiations (방사선 조사량에 따른 인체 정상 림파구의 미세핵 발생빈도)

  • Kim Sung-Ho;Cho Chul-Koo;Kim Tae-Hwan;Chung In-Yong;Yoo Seong-Yul;Koh Kyoung-Hwan;Yun Hyong-Geun
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.35-42
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    • 1993
  • The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with $\gamma$-rays and neutrons in the 5 dose ranges was studied for a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-dependent increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model, frequencies per 1000 CB cells were ($0.31{\pm}0.049$)D+($0.0022{\pm}0.0002)D^2+(13.19{\pm}1.854) (r^2=1.000,\;X^2=0.7074,\;p=0.95$) following $\gamma$ irradiation, and ($0.99{\pm}0.528$)\;D+(0.0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309)\;(r^2=0.996,\;X^2=7.6834,\;p=0.11) following neutrons irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with $\gamma$-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was $2.37{\pm}0.17$. Since the MN assay is simple and rapid, it may be a good tool for evaluating the $\gamma$-ray and neutron response.

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Isolation and Characterization of mas1+ of Schizosaccharomyces pombe, a Homologue of Human CIP29/Hcc-1 Involved in the Regulation of Cell Division (세포분열에 관여하는 인간의 CIP29/Hcc1 유전자와 상동성을 가지는 분열형 효모의 새로운 유전자 mas1+의 특성분석)

  • Cha, Jae-Young;Shin, Sang-Min;Ha, Se-Eun;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1666-1677
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    • 2011
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

A Cancer-specific Promoter for Gene Therapy of Lung Cancer, Protein Regulator of Cytokinesis 1 (PRC1) (폐암의 유전자 치료법을 위한 암특이적인 PRC1 프로모터)

  • Cho, Young-Hwa;Yun, Hye-Jin;Kwon, Hee-Chung;Kim, Hee-Jong;Cho, Sung-Ha;Kang, Bong-Su;Kim, Yeun-Ju;Seol, Won-Gi;Park, Kee-Rang
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1395-1399
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    • 2008
  • We have recently reported the PRC1 promoter as a promoter candidate to control expression of transcriptionally targeted genes for breast cancer gene therapy. We tested whether the PRC1 promoter could be also applied for the lung cancer gene therapy. In the transient transfection assay with naked plasmids containing the luciferase fused to the PRC1 promoter, the promoter showed little activity in the normal lung cell line, MRC5. However, in the lung cancer A549 cells, PRC1 showed approximately 30-fold activation which was similar to the survivin promoter, the gene whose promoter has been already reportedas a candidate for the gene therapy of lung cancer. In viral systems, the PRC1 promoter showed approximately 75% and 66% of transcriptional activity compared to the CMV promoter in the adeno-associated virus (AAV) and the adenovirus (AV) systems, respectively. However, the PRC1 promoter in either AAV or AV showed approximately 20% activity compared to the CMV promoter in the normal lung cells. In addition, human lung tumor xenograft mice showed that the PRC1 promoter activity was as strong as the CMV activity in vivo. Taken together, these results suggested that PRC1 might be a potential promoter candidate for transcriptionally targeted lung cancer gene therapy.

Nuclear Anomalies, Chromosomal Aberrations and Proliferation Rates in Cultured Lymphocytes of Head and Neck Cancer Patients

  • George, Alex;Dey, Rupraj;Bhuria, Vikas;Banerjee, Shouvik;Ethirajan, Sivakumar;Siluvaimuthu, Ashok;Saraswathy, Radha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1119-1123
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    • 2014
  • Head and neck cancers (HNC) are extremely complex disease types and it is likely that chromosomal instability is involved in the genetic mechanisms of its genesis. However, there is little information regarding the background levels of chromosome instability in these patients. In this pilot study, we examined spontaneous chromosome instability in short-term lymphocyte cultures (72 hours) from 72 study subjects - 36 newly diagnosed HNC squamous cell carcinoma patients and 36 healthy ethnic controls. We estimated chromosome instability (CIN) using chromosomal aberration (CA) analysis and nuclear level anomalies using the Cytokinesis Block Micronucleus Cytome Assay (CBMN Cyt Assay). The proliferation rates in cultures of peripheral blood lymphocytes (PBL) were assessed by calculating the Cytokinesis Block Proliferation Index (CBPI). Our results showed a significantly higher mean level of spontaneous chromosome type aberrations (CSAs), chromatid type aberration (CTAs) dicentric chromosomes (DIC) and chromosome aneuploidy (CANE UP) in patients (CSAs, $0.0294{\pm}0.0038$; CTAs, $0.0925{\pm}0.0060$; DICs, $0.0213{\pm}0.0003$; and CANE UPs, $0.0308{\pm}0.0035$) compared to controls (CSAs, $0.0005{\pm}0.0003$; CTAs, $0.0058{\pm}0.0015$; DICs, $0.0005{\pm}0.0003$; and CANEUPs, $0.0052{\pm}0.0013$) where p<0.001l. Similarly, spontaneous nuclear anomalies showed significantly higher mean level of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) among cases (MNi, $0.01867{\pm}0.00108$; NPBs, $0.0156{\pm}0.00234$; NBUDs, $0.00658{\pm}0.00068$) compared with controls (MNi, $0.00027{\pm}0.00009$; NPBs, $0.00002{\pm}0.00002$; NBUDs, $0.00011{\pm}0.00007$).The evaluation of CBPI supported genomic instability in the peripheral blood lymphocytes showing a significantly lower proliferation rate in HNC patients ($1.525{\pm}0.005552$) compared to healthy subjects ($1.686{\pm}0.009520$) (p<0.0001). In conclusion, our preliminary results showed that visible spontaneous genomic instability and low rate proliferation in the cultured peripheral lymphocytes of solid tumors could be biomarkers to predict malignancy in early stages.

Embryology of Jeffersonia dubia Baker et S. Moore (Berberidaceae) and comparison with allied genera (깽깽이풀의 발생과 근연속간 비교)

  • Ghimire, Balkrishna;Heo, Kweon
    • Korean Journal of Plant Taxonomy
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    • v.42 no.4
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    • pp.260-266
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    • 2012
  • Because the embryological features of Jeffersonia dubia are poorly understood, we conducted the first embryological study comparing it to other related genera of Berberidaceae. Important embryological features of J. dubia are as follows: the anther is tetrasporangiate, anther wall formation confirms basic type, glandular tapetum cells are two nucleate, the epidermis persistent, and the endothecium develops fibrous thickenings, anther dehiscence by two valves, meiosis in a microspore mother cell is accompanied by simultaneous cytokinesis, microspore tetrads are usually tetrahedral, pollen grains two cells at the time of anthesis. The ovule is bitegmic, anatropous and crassinucellate, archesporium single celled, development of the embryo sac Polygonum type, a mature embryo sac is ellipsoidal in shape. Endosperm formation is of Nuclear type and embryogeny Onagrad type. Seeds are arillate and seed coat exotestal type. Embryological comparisons showed that Jeffersonia resemble to Epimedium and Vancouveria rather than Berberis and Mahonia in some features, like as number of tapetal cells, cytokinesis in meiosis, and thickness of exotesta. It also resembles to Gymnospermium in mode of anther wall formation, number of tapetal cells, formation of nucellar cap, and nature of antipodal cells. Nevertheless, Jeffersonia and Gymnospermium differ from several other embryological features and molecular data too. Therefore, embryological evidences support that Jeffersonia is closely related with Epimedium and Vancouveria.

Studies on Intracellular Functions of the mas3 Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서 mas3 유전자의 세포내 기능 연구)

  • Hwang Mi Ra;Cha Jae Young;Shin Sang Min;Park Jong Kun
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.124-131
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    • 2005
  • The regulation of gene expression plays an important rolet in cell cycle controls. In this study, a novel $mas3^+$ (mitosis associated protein) gene, a homolog of human SMARCADl, was isolated and characterized from a fission yeast Schizosaccharomyces pombe. The overall homology between the helicase proteins of the two species is $87\%$. This DEAD/H box-containing molecule has seven highly conserved sequence regions that allow us to place it in the SNF2 family of the helicase superfamily. Knock-out cell of $mas3^+$ gene was constructed using kanMX6 as a selection marker. Survival of mas3 null mutant exposed to UV or MMS was similar to those of wild type cells. $mas3^+$ expression was lowest at $G_2$ and gradually increased. Cytokinesis of mas3 null mutant was abnormal at $26^{\circ}C\;and\;35^{\circ}C$ and a large number of multi-septate cells were produced. These results indicate that the $mas3^+$ is involved in cytokinesis and cell shape control.

Incidence of micronuclei in lymphocytes of pig in the high background radiation area (Cheongwon-gun and Boeun-gun) (국내 자연 방사선 고준위 지역(청원군, 보은군) 사육 돼지의 림프구 미소핵 발생 평가)

  • Lee, Hae-June;Kang, Chang-Mo;Kim, Se-Ra;Lee, Jin-Hee;Kim, Joong-Sun;Kim, Jong-Choon;Kim, Ill-Hwa;Kim, Tae-Hwan;Ryu, Si-Yun;Jo, Sung-Kee;Choi, Soo-Yong;Kim, Sung-Ho
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.469-475
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    • 2005
  • Cytogenetic and hematological analysis was performed in peripheral blood obtained from pigs bred in the high background radiation areas (HBRA) (Cheongwon-gun and Boeun-gun) and a control area. The frequencies of gamma-ray induced micronuclei (MN) in the cytokinesis-blocked (CB) lymphocytes at several doses were measured in three pigs. An estimated dose of radiation was calculated by a best fitting linear-quadratic model based on the radiation-induced MN formation from the swine lymphocytes exposed in vitro to radiation over the range from 0 mGy to 1,969 mGy. The measurements performed after irradiation showed dose-related increases in the MN frequency in each donors. The results were analyzed using a linear-quadratic model with a line of best fit of $y=0.0005404D^2+0.04237D+0.00833$ [y = number of MN/cytokinesis-blocked (CB) cells and D = irradiation dose in Gy]. MN rates per 1,000 CB lymphocytes of pig from the HBRA (Cheongwon-gun, Boeun-gun) and the control area were $6.70{\pm}2.36$, $9.00{\pm}3.50$ and $11.00{\pm}2.98$, respectively. The MN frequencies of CB lymphocytes from pigs bred in three areas means that the values are within the background variation in this experiment. The MN frequencies and hematological values were similar regardless of whether the pigs were bred in the HBRA or the control area.