• Title/Summary/Keyword: Nucleus condensation

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Nucleolar translocalization of GRA10 of Toxoplasma gondii transfectionally expressed in HeLa cells

  • Ahn, Hye-Jin;Kim, Sehra;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.45 no.3
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    • pp.165-174
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    • 2007
  • Toxoplasma gondii GRA10 expressed as a GFP-GRA10 fusion protein in HeLa cells moved to the nucleoli within the nucleus rapidly and entirely. GRA10 was concentrated specifically in the dense fibrillar component of the nucleolus morphologically by the overlap of GFP-GRA10 transfection image with IFA images by monoclonal antibodies against GRA10 (Tg378), B23 (nucleophosmin) and C23 (nucleolin). The nucleolar translocalization of GRA10 was caused by a putative nucleolar localizing sequence (NoLS) of GRA10. Interaction of GRA10 with TATA-binding protein associated factor 1B (TAF1B) in the yeast two-hybrid technique was confirmed by GST pull-down assay and immunoprecipitation assay. GRA10 and TAF1B were also co-localized in the nucleolus after co-transfection. The nucleolar condensation of GRA10 was affected by actinomycin D. Expressed GFP-GRA10 was evenly distributed over the nucleoplasm and the nucleolar locations remained as hollows in the nucleoplasm under a low dose of actinomycin D. Nucleolar localizing and interacting of GRA10 with TAF1B suggested the participation of GRA10 in rRNA synthesis of host cells to favor the parasitism of T. gondii.

An Experimental Study on the Characteristics of Direct Photoelectric Charging (직접 광대전의 대전특성에 관한 실험적 연구)

  • Lee, Chang-Sun;Kim, Yong-Jin;Kim, Sang-Sao
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.24 no.6
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    • pp.753-759
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    • 2000
  • Photoelectric charging is a very efficient way of charging small particles. This method can be applied to combustion measurement, electrostatic precipitator, metal separation and control of micro-contamination. To understand the photoelectric charging mechanism, particle charging of silver by exposure to ultraviolet is investigated in this study. Average charges and charge distributions are measured at various conditions, using two differential mobility analyzers, a condensation nucleus counter, and an aerosol electrometer. The silver particles are generated in a spark discharge aerosol generator. After that process, the generated particles are charged in the photoelectric charger using low-pressure mercury lamp that emits ultraviolet having wavelength 253.7 nm. The results show that ultra-fine particles are highly charged by the photoelectric charging. The average charges linearly increase with increasing particle size and the charge distribution change with particle size. These results are discussed by comparison with previous experiments and proposed equations. It is assumed that the coefficient of electron emission probability is affected by initial charge. The results also show that the charge distribution of a particle is dependent on initial charge. Single changed particle, uncharged particle and neutralized particle are compared. The differences of charge distribution in each case increase with increasing particle size.

An Experimental Study on Enhancement of the Filter Efficiency by the Image Effect of Charged Particle (대전된 입자의 영상효과에 의한 필터효율 향상에 관한 실험적 연구)

  • Lee, Chang-Sun;Jeong, Hae-Young;Kim, Sang-Soo
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.24 no.6
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    • pp.760-768
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    • 2000
  • Filter efficiency of electrically charged particle in uncharged fibrous filter was measured. In previous studies, the effect of charged particle on filter efficiency was investigated but there was difficulty in measuring of image effect that is appeared at the charged small particle. We could easily measure the image effect with charging small particles by photoelectric charging. The spark discharge aerosol generator and a differential mobility analyzer (DMA) were used to generate sub-micron monodisperse particles (${\leq}200$ nm). The generated particles were charged in photoelectric charging process using ultraviolet lamp and electric field. The filter efficiency of the charged particles, classified by another DMA, was measured in filter tester using a condensation nucleus counter (CNC) as function of particle diameter, particle charge and airflow velocity. It is shown that the filter efficiency increases with increasing charge number of the particle and is affected by particle size and flow velocity. Single fiber filter efficiency mainly depends on image force parameter and peclet number. The peclet number was not considered at previous other papers. We propose a modi fied experimental correlation as function of image force parameter and peclet number.

Heat-Killed Enterococcus faecium KU22001 Having Effective Anti-Cancer Effects on HeLa Cell Lines at a Lower Temperature

  • Jun-Su Ha;Na-Kyoung Lee;Hyun-Dong Paik
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.902-910
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    • 2024
  • The anti-cancer effects of heat-killed Enterococcus faecium KU22001 (KU22001), KU22002, and KU22005 isolated from human infant feces were investigated. The anti-proliferative activity of these strains against various cancer cell lines was evaluated using the MTT assay. To determine the production of exopolysaccharides (EPS) with potential anti-cancer effect, ethanol precipitation and phenol-sulfuric acid method was used with the cell free supernatant of strains grown at 25℃ or 37℃. The EPS yield of E. faecium strains was higher at 25℃ than at 37℃. Among these E. faecium strains, KU22001 grown at 25℃ was associated with the highest bax/bcl-2 ratio, effective apoptosis rate, cell cycle arrest in the G0/G1 phase, and condensation of the nucleus in the cervical cancer HeLa cell line. In conclusion, these results suggest that KU22001 can be beneficial owing to the anti-cancer effects and production of functional materials, such as EPS.

Survival and Histological Change of Integumentary System of the Juvenile Sea Cucumber, Apostichopus japonicus Exposed to Various Salinity Concentrations (염분 농도에 따른 어린 돌기해삼, Apostichopus japonicus의 생존 및 피부계의 조직학적 변화)

  • Kim, Tae-Ik;Park, Min-Woo;Cho, Jae-Kwon;Son, Maeng-Hyun;Jin, Young-Guk
    • Journal of Fisheries and Marine Sciences Education
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    • v.25 no.6
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    • pp.1360-1365
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    • 2013
  • Juvenile sea cucumber, Apostichopus japonicus(wet weight $1.0{\pm}0.2g$, n=250) acclimated to environmental conditions($17^{\circ}C$, 33 psu) during 4 weeks for observation of survival and structural variation of integumentary system according to various salinity concentration. After that, it was exposed 0, 5, 10, 15, 20, 25, 30, 35, 40 psu in 96h, and recovered during 7 days. In this results, The 96-h lethal concentration(LC50) for sea cucumber was a salinity of 21.05 psu. Integumentary system of the sea cucumber exposed to below the salinity of 20 psu mainly observed thickness reduction of epidermal layer, nucleus condensation of epithelial cells, decreased of mucous cells and loose arrangement of connective tissue in dermal layer. Integumentary system of the sea cucumber exposed a salinity of 40 psu mainly observed nucleus hypertrophy of epithelial cells, increase of mucous cells and tight arrangement of connective tissue in dermal layer. Also, observed secretory cells showing the alcian blue(pH 2.5) positive. During 96-h exposure, this cells decreased after increase below the salinity of 20 psu but increased in the salinity of 40 psu compared with 25-35 psu.

Ultrastructural Study of Chitosan on the Liver of Cadmium-Administrated Mouse (Chitosan이 Cadmium을 투여한 흰쥐의 간에 미치는 미세구조적 연구)

  • Yoon, Jung-Sik;Roh, Young-Bok
    • Applied Microscopy
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    • v.27 no.3
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    • pp.265-279
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    • 1997
  • This study aims to demonstrate the effect of chitosan, one of the natural chelator, on the ultrastructural changes in the mouse liver caused by cadmium. A total of 60 healthy ICR that weighted 30 gm $({\pm}2gm)$ was used for experiment. The experimental group was divided into three groups; group A, B, and C. The group A and B administrated cadmium (4.0 mg/kg) to the intraperitoneal after pretreated with chitosan (0.5% solution) for 30 and 7 days, respectively. Each group was observed at 12, 24, 48, 72 hours and one week after injected cadmium. The results were as follows: 1. Group A The nuclear membrane and the chromatin were normal shapes at overall the time. The inner and outer membranes of the mitochondria damaged a little but almost normal in shapes. And electron-density showed slightly compacted. some enlarged rER (rough endoplasmic reticulum) showed at 12 hours. At 48 hours, typical lamellae of the rER were reformed, and a lot of transvesicles observed around them. To 48 hours, sER (smooth endoplasmic reticulum) was slightly dilated. From 72 hours, sER rehalizated in normal shape. 2. Group B Nuclear membranes were rounded-shape and chromatin showed evenly. To 72 hours, a lot of mitochondria observed around rER and development of cristae showed weakly. But at one week, cristae were clear and electron-density of matrix showed high. At 72 hours lamellae of rER showed some broken, but were reformed at one week. Also at one week, glycogen granules evenly showed over cytoplasm. 3. Group C At 12 hours, Nucleus showed the condensation of nuclear membrane and clear condensation at 24 hours. However, nuclear membrane had a slightly rounded-shape from 72 hours. From 12 hours to the one week, mitochondria showed the dilation of inner cavity and weak development of cristae. Also electron-density of matrix was a little low. Occasionally, destruction of inner and outer membrane observed at one week. The dilation of cisternae and destruction of lamellae of rER showed from 12 to 48 hours. From 72 hours, rER showed slightly dilated only. And lamella observed at one week. In sER, dilation of inner cavity was observed during whole period. These results suggest that chitosan attenuates the toxic effect of the cadmium in the mouse liver.

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Thymocyte Apoptosis Induced by Cyclophosphamide in Rats (랫드에서 cyclophosphamide에 의해 유발된 흉선세포의 apoptosis)

  • 구현옥;권창희;조준형;정상희;박신자;김윤배;양재만;이영순
    • Toxicological Research
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    • v.13 no.1_2
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    • pp.39-48
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    • 1997
  • Cyclophosphamide(25, 50 or 100 mg/kg), orally administered to male Sprague-Dawley rats, caused a time- and dose-dependent thymic atrophy. In the light microscopic examination of the atrophic thymus, thymocytes with condensed or fragmented nucleus were multifocally observed in the cortical region, started to increase 8 hr after CPA treatment and reached to the maximal level at 16 hr, although such cells were not seen after 48 hr when the severe depletion of thymocytes were marked. In agarose gel electrophoresis to analyze the DNA changes, DNA extracted from atrophic thymus showed a oligonucleosomal laddering at the corresponding time to morphological changes. In an additional supportive experiment, thymocytes showing morphological changes, nuclear condensation or apoptotic body, exhibited a positive reaction to immunoperoxidase staining using in situ apoptosis detection kit. Separately, agarose gel electrophoresis of DNA from bone .marrow cells was performed to investigate the involvement of bone marrow cells in the process of thymocyte apoptosis. Although DNA laddering was slightly increased 2 and 4 hr after treatment, no clear correlation was inferred. Taken togather, it is concluded that thymocytes showing morphological changes in thymic atrophy induced by cyclophosphamide administration represent an apoptosis having biochemical nature of programmed cell death.

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Effect of Disk Rotational Speed on Contamination Nano Particles Generated in a Hard Disk Drive (하드 디스크 드라이브 회전수 변화가 드라이브 내 나노 오염 입자 발생에 미치는 영향)

  • Lee, Dae-Young;Hwang, Jung-Ho;Bae, Gwi-Nam
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.28 no.8 s.227
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    • pp.976-983
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    • 2004
  • In high-density hard disk drives, the slider should be made to fly close to the magnetic recording disk to generate better signal resolution and at an increasingly high velocity to achieve better data rate. The slider disk interaction in CSS (contact-start-stop) mode is an important source of particle generation. Contamination particles in the hard disk drive can cause serious problems including slider crash and thermal asperities. We investigated the number and the sizes of particles generated in the hard disk drive, operating at increasing disk rotational speeds, in the CSS mode. CNC (condensation nucleus counter) and PSS (particle size selector) were used for this investigation. In addition, we examined the particle components by using SEM (scanning electron microscopes), AES (auger electron spectroscopy), and TOF-SIMS (time of flight-secondary ions mass spectrometry). The increasing disk rotational speed directly affected the particle generation by slider disk interaction. The number of particles that were generated increased with the disk rotational speed. The particle generation rate increased rapidly at motor speeds above 8000 rpm. This increase may be due to the increased slider disk interaction. Particle sizes ranged from 14 to 200 nm. The particles generated by slider disk interaction came from the lubricant on the disk, coating layer of the disk, and also slider surface.

Cell Surface Interaction with Expression of Fas Ligand Mediates Prolactin-Induced Apoptosis In Rat Luteal Cell Culture (Rat 황체세포 배양에 있어서 Prolactin에 의한 황체퇴행 및 Fas Ligand의 발현)

  • 장규태;박미령;선동수;윤창현
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.179-190
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    • 1998
  • Prolactin (PRL) surge in cycling rats at proestrous afternoon has previously been reported as an inducer of apoptotic cell death of luteal cells. This death-inducing action of PRL seeins unusual, because PRL can he categorized as a cell-survival factor, if other known physiological functions of PRL are taken into account. In this study, the apoptotic action of PRL was assessed in cultured cells prepared from rat luteal tissue and underlying molecular /cellular mechanism of PRL-induced luteolysis was analyzed. The latest crop of corpora lutea (CLs) were enucleated from rat ovaries at 18:00 h on the proestrous day before the next ovulation. Donor rats were pretreated with CB154, a dopamine agonist, in order to he exempted from the endogenous PRL surge. The harvested GLs were dispersed and cultured with or without PRL (2$\mu$g /ml) for 24 or 48 h. An addition of PRL to the culture medium changed the parameters indicative of cell death via apoptosis: a decrease in cell viability (MTT) and an increase in chromatin condensation. Most of the DNA breakdown in nuclei induced by PRL occurred in steroidogenic cells which were identified by 3$\beta$-HSD activity staining, and the number of 3$\beta$-HSD-positivecells were significantly decreased. Interestingly, most of the cells with an apoptotic nucleus adhered to one or more intact and seemingly non-steroidogenic cells. Because the expression of Fas has heen shown to be abundant in murine ovary, and Fas is known to have an exact physiological role in occurrence of apoptotic cell death, the membrane form-Fas ligand (rnFasL) was quantified in the cell lysate. An addition of PRL increased expression of mFasL. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, in place of PRL, enhanced the apoptotic parameters. Cumulatively, the apoptotic PRL action was addressed to cells unknown than steroidogenic lute~ cells. The most prohable candidate for the direct target cells is Tcells in the luteal tissue that can express mFasL in response to PRL.

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Nuclear Modeling and Developmental Potential of Bovine Somatic Nuclear Transfer Embryos Cloned by Two Different Activation Methods

  • Jeon, Byeong-Gyun;Rho, Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.105-113
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    • 2011
  • The present study investigated the nuclear remodeling, development potential with telomerase activity and transcription level of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos using two different fusion and activation methods. Female adult fibroblasts were injected into perivitelline space of in vitro matured oocytes. The oocyte-nucleus complexes were fused and followed by immediately either activated (Group 1), or activated at 1 h post-fusion (hpf) (Group 2), respectively. The incidence of normal premature chromosome condensation (PCC) at 1 hpf was slightly increased in the Group 2, compared to those of Group 1, but there was no significant (p<0.05) difference. The incidence of normal pronucleus (PN) and chromosome spread at 5 and 18 hpf were significantly (p<0.05) higher in the Group 2 than those of Group 1. The cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cell numbers were significantly (p<0.05) higher in the Group 2, compared to those of Group 1. Level of telomerase activity was significantly (p<0.05) higher in the SCNT blastocysts of Group 2, compared to those of Group 1. Transcript levels of HPRT, MeCP2 and XIST were not significantly (p<0.05) different between blastocysts of Group 1 and 2. However, transcript level of ANT3, RPS4X, XIAP and ZFX were significantly (p<0.05) up-regulated in the SCNT blastocysts of Group 2, compared to those of Group 1. Taken together, it is concluded that oocyte activation at 1 hpf induces the enhanced developmental potential by efficient nuclear remodeling and subsequent facilitation of the nuclear reprogramming of bovine SCNT embryos.