• Title/Summary/Keyword: Cell Cycle Stage

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PLGA-Loaded Gold-Nanoparticles Precipitated with Quercetin Downregulate HDAC-Akt Activities Controlling Proliferation and Activate p53-ROS Crosstalk to Induce Apoptosis in Hepatocarcinoma Cells

  • Bishayee, Kausik;Khuda-Bukhsh, Anisur Rahman;Huh, Sung-Oh
    • Molecules and Cells
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    • v.38 no.6
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    • pp.518-527
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    • 2015
  • Controlled release of medications remains the most convenient way to deliver drugs. In this study, we precipitated gold nanoparticles with quercetin. We loaded gold-quercetin into poly(DL-lactide-co-glycolide) nanoparticles (NQ) and tested the biological activity of NQ on HepG2 hepatocarcinoma cells to acquire the sustained release property. We determined by circular dichroism spectroscopy that NQ effectively caused conformational changes in DNA and modulated different proteins related to epigenetic modifications and c ell cycle control. The mitochondrial membrane potential (MMP), reactive oxygen species (ROS), cell cycle, apoptosis, DNA damage, and caspase 3 activity were analyzed by flow cytometry, and the expression profiles of different anti- and pro-apoptotic as well as epigenetic signals were studied by immunoblotting. A cytotoxicity assay indicated that NQ preferentially killed cancer cells, compared to normal cells. NQ interacted with HepG2 cell DNA and reduced histone deacetylases to control cell proliferation and arrest the cell cycle at the sub-G stage. Activities of cell cycle-related proteins, such as $p21^{WAF}$, cdk1, and pAkt, were modulated. NQ induced apoptosis in HepG2 cells by activating p53-ROS crosstalk and induces epigenetic modifications leading to inhibited proliferation and cell cycle arrest.

Production of Cloned Embryos and Animals following Regulation of Cell Cycle of Donor Nucleus and Type of Recipient Cytoplasm (토끼에서 공핵란의 세포주기 조절과 수핵란의 세포질 상태에 따른 핵이식 수정란의 체외 발달과 복제동물의 생산)

  • 박충생;전병균;하란조;윤희준;곽대오;이효종;최상용
    • Journal of Embryo Transfer
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    • v.12 no.3
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    • pp.259-267
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    • 1997
  • To improve the efficiency of production of cloned embryos and animals by nuclear transplantation in the rabbit, the effect of cell cycle of donor nuclei and type of recipient cytoplasm on the in vitro developmental potential and production efficiency of offspring was determined. The embryos of 16-cell stage were collected from the mated does at 48h post-hCG injection and they were synchronized to G$_1$ phase of 32-cell stage. The oocytes collected at 14h post-hCG injection were freed from cumulus cells and then enucleated. One group of the enucleated cytoplasms was activated by electrical stimulation prior to injection of donor nucleus, and the other group was not pre-activated. The separated G$_1$phase blastomeres of 32-cell stage embryos were injected into the perivitelline space of recipient cytoplasms. After culture for 20h post-hCG injection, the nuclear transplant oocytes were electrofused and activated by electrical stimulation and the fused nuclear transplant embryos were co-cultured for 120h and the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 dye and their blastomeres were counted. Some of the nuclear transplant embryos developed in vitro to 2- to 4-cell stage were transferred into the oviducts of synchronized recipient does. The electrofusion rate was similar between the types of donor nuclei and recipient cytoplasms used. However, the nuclear transplant embryos using G$_1$ phase donor nuclei were developed to blastocyst at higher rate(60.3%) than those using S phase ones(24.7%). Also, when non-preactivated oocytes were used as recipient cytplasms, the develop-mental rates of nuclear transplant embryos to blastocysts were significantly(P< 0.05) higher(57.1%) than those using preactivated ones(20.8%). The cell counts of nuclear transplant embryos developed to blastosyst stage were increased signficantly(P<0.05) more in the non-preactivated recipient cytoplasm(163.7 cells), as compared whit the preactivated recipient cytoplasm(85.4 cells), A total of 49 nuclear transplant embryos were tranferrid into 5 recipient does, of which two offsprings were produced from a foster mother 31 days after embryo transfer. these results showed that the blastomeres of G1 phase and non-preactivated oocytes might be utillzed efficiently as donor nuclei and recipient cytoplasms in the nuclear transplant procedure, thought the offspring production remained still low.

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Effects of Abiotic Stresses on Cell Cycle Progression in Tobacco BY-2 Cells

  • Jang, Su Jin;Shin, Sung Hae;Yee, Sung Tae;Hwang, Baek;Im, Kyung Hoan;Park, Ky Young
    • Molecules and Cells
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    • v.20 no.1
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    • pp.136-141
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    • 2005
  • Mild stresses such as high temperature ($30^{\circ}C$) or a low $H_2O_2$ concentration induced transient cell cycle arrest at G1/S or G2/M depending on the cell cycle stage at which the stress was applied. When stresses were introduced during G0 or G1, the G1/S checkpoint was mainly used; when stresses were introduced after S phase, G2/M was the primary checkpoint. The slowing of cell cycle progression was associated with transient delays in expression of A-, B-, and D-type cyclins. The delay in expression of NtcycA13, one of the A-type cyclins, was most pronounced. The levels of expression of Ntcyc29 (a cyclin B gene) and of CycD3-1 differed most depending on the applied stress, suggesting that different cellular adjustments to mild heat and a low concentration of $H_2O_2$ are reflected in the expression of these two cyclins.

Seminiferous Epithelium Cycle in Bombina orientalis

  • Yi, Min-Joo;Lee, Jung-Hun
    • Development and Reproduction
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    • v.19 no.1
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    • pp.1-10
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    • 2015
  • The purpose of the present study was to examine the seminiferous epithelium cycle of Bombina orientalis using a light microscope. The cycle was divided into a total of 10 stages, according to the morphological characteristics of the cells. The spermatogenetic cells included primary spermatogonia, secondary spermatogonia, primary spermatocytes, secondary spermatocytes, spermatid and sperm. At stage I, the primary spermatogonia was located closer to basal lamina of the seminiferous tubule without spermatocyst formations. Especially at the stage II, the secondary spermatogonia were located in the spermatocyst. The primary and secondary spermatocytes were found from stages III to VI. The secondary spermatocytes were smaller in size than the primary spermatocytes, but they had thicker nucleoplasm and smaller nuclei. The round-shaped, early sperm cells were formed in stage VII, and further divided at stage VIII to have more concentrated nucleoplasm before division to matured sperm cells. At stage X, the matured sperm cells emerged from the spermatocyst. Considering the above results, this study presented the special characteristics in the generation and type of sperm formation. The germ cell formation occurred in various stages, like the perspectives of Franca et al (1999), ultimately, providing taxonomically useful information.

Gametogenic Cycle and Fine Structure of Ripe Germ Cells in the Pacific Oyster, Crassostrea gigas on the South Coast of Korea

  • Choi Youn Hee;Kim Tae Ik;Hur Young Baek;Go Chang-Soon;Chang Young Jin
    • Fisheries and Aquatic Sciences
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    • v.6 no.2
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    • pp.51-58
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    • 2003
  • The gonadal development and the gametogenic cycle and the fine structure of ripe germ cells of the cultured Pacific oyster, Crassostrea gigas were investigated using oysters monthly collected from the southern coast of Korea from October 2000 to September 2001. Monthly changes in the condition index were similar to that of meat weight rate and the highest value was observed in between April and May, and the lowest value in August. The external colors of the testis and the ovary were milky white and yellowish, respectively. The spawning period of the Pacific oyster was continued from May to September, with a peak in July. The gametogenic cycle could be classified into five successive stages: multiplicative stage (December to March), growing stage (March and April), mature stage (April to June), spawning stage (June to August) and resting stage (August to January). Variety of egg yolk granules, lipid granules, mitochondria, and endoplasmic reticula were observed in cytoplasm of ripe oocyte. The spermatozoon consisted of the head, middle piece and tail; including cap-shaped acrosome with domed structure, elliptical shaped nucleus, four mitochondria, two centrioles and flagellum.

Spermatogenesis and Reproductive Cycle in Male Spisula sachalinensis (Bivalvia: Mactridae) of Korea

  • Lee, Ki-Yong;Chung, Ee-Yung;Lee, Jeong-Yong
    • The Korean Journal of Malacology
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    • v.24 no.1
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    • pp.1-10
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    • 2008
  • Spermatogenesis and the reproductive cycle in male Spisula sachalinensis were investigated by cytological and histological observations. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves in that it contains a short midpiece with four mitochondria surrounding the centrioles. But spermatozoon of this species has not axial rod and satellite body in the midpiece. The morphologies of the sperm nucleus type and the acrosome shape of this species have a globe-shape type and modified cap-like shape, respectively. The spermatozoon is approximately $40-45{\mu}m$ in length including the sperm nucleus length (about $1.35{\mu}m$), acrosome length (about $1.50{\mu}m$) and tail flagellum. The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9+2 structure. The spawning period of these species lasts from June to July, and the main spawning occurs in July when seawater temperatures are greater than $20^{\circ}C$. The male reproductive cycle of this species can be categorized into five successive stages: early active stage (October to January), late active stage (February to April), ripe stage (April to June), partially spawned stage (June and July), and spent/inactive stage (August to September).

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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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Host-Parasite System in a Red Tide Dinoflagellate Prorocentrum minimum: (1) Life Cycle Stages of the Parasitic Dinoflagellate Amoebophrya sp. (적조생물 Prorocentrum minimum의 숙주-기생자 배양체: (1) 기생성 와편모류 Amoebophrya sp.의 생활사 단계)

  • 김영길;박명길;이원호
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.7 no.4
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    • pp.2221-2221
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    • 2002
  • The first laboratory culture of host-parasite system of Prorocentrum minimum- Amoebophrya sp. was established by single cell isolation method. Here, we report the life cycle stages of the parasitic dinoflagellate. Amoebophrya sp. of the red tide dinoflagellate P. minimum as observed by light and epifluorescence microscopy. Infections developed inside the nucleus of P. minimum. The trophont developed to occupy almost all the intracellular space of the host at its late stage. The fully developed trophont finally ruptured through the host cell. “Vermiform stage”, the free-swimming extracellular lift cycle stage is followed by another stage for the sudden release of many individual dinospores. Our laboratory strain of the host-parasite system for P. minimum, a causative species fur the huge red tides in spring and summer in Korean coastal waters, could be a useful living material for the in situ biological control of harmful algal blooms.

Host-Parasite System in a Red Tide Dinoflagellate Prorocentrum minimum:(1) Life Cycle Stages of the Parasitic Dinoflagellate Amoebophrya sp. (적조생물 Prorocentrum minimum의 숙주-기생자 배양체: (1) 기생성 와편모류 Amoebophrya sp.의 생활사 단계)

  • 김영길;박명길;이원호
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.7 no.4
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    • pp.221-225
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    • 2002
  • The first laboratory culture of host-parasite system of Prorocentrum minimum- Amoebophrya sp. was established by single cell isolation method. Here, we report the life cycle stages of the parasitic dinoflagellate. Amoebophrya sp. of the red tide dinoflagellate P. minimum as observed by light and epifluorescence microscopy. Infections developed inside the nucleus of P. minimum. The trophont developed to occupy almost all the intracellular space of the host at its late stage. The fully developed trophont finally ruptured through the host cell. “Vermiform stage”, the free-swimming extracellular lift cycle stage is followed by another stage for the sudden release of many individual dinospores. Our laboratory strain of the host-parasite system for P. minimum, a causative species fur the huge red tides in spring and summer in Korean coastal waters, could be a useful living material for the in situ biological control of harmful algal blooms.

Physiological studies on cell division by the technique of synchronous culture of chlorella (II) (클로렐라의 동조배양법에 의한 세포분열의 생리학적 연구 2)

  • 이영녹;심웅섭
    • Korean Journal of Microbiology
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    • v.7 no.1
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    • pp.10-21
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    • 1969
  • The effect of glucose and 2-thiobarbituric acid on the biosynthesis of cell constituents such as protein, carbohydrate, DNA, RNA, phospholipid and PCA-soluble phosphate compounds in Chlorella duing the life cycle was measured, and the changes in the content of these main cellular components of the algal cell were analyzed in connection with the nuclear and cytoplasmic divison. In the normal autotrophic synchronous culture the contents of protein, RNA, and DNA in the cell showed a chracteristic changes according to the progress of cell development, increasing more or less throughout all the life cycle. The synthesis of protein is more prominent in the division period nad that of DNA is more active in the ripening period, while the synthesis of RNA is more rapid in the growing and ripening periods than other developmental stages. The period of division cycle was little affected by glucose in the medium, although the synchrony of the growth and cellular division was disturbed and the n value increased. The cotents of protein, carbohydrate, RNA nad DNA of the cell were increased by the glucose treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment. The synthesis of protein, carbohydrate, DNA, RNA and phospholipid of the cell was also retarded by 2-thiobarbituric acid. In the autotrophic, mixotrophic and 2-thiobarbituric acid-treated cultures, each having different mode cytoplasmic division, a common general schema occurring in the cell during the life cycle may be drawn as follows. The ratio of RNA to protein attains maximum value in the $L_1$-cell stage prior to the nuclear division and thereafter decreases during the periods of ripening and division. The ratio of PCA-soluble phosphate compounds to protein increased from the begining of the culture to $L_4$-cell stage successively and thereafter decreased gradually during the division period, while the ratio of protein to DNA kept almost constant up to the division period and thereafter increased during the division period. Therefore, it is presumed that the increase in the ratio of RNA to protein is to be an inducer of nuclear division and that the cytoplasmic division is induced by the increase in the ratio of protein to DNA.

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