• Title/Summary/Keyword: Cell Disruption

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Effect of Chlorambucil on the Ultrastructure of Leydig Cell in Male Mouse (수컷생쥐의 Leydig Cell 미세구조에 대한 Chlorambucil의 영향)

  • Kim, Douk-Hoon;Park, Won-Hak
    • Applied Microscopy
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    • v.18 no.2
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    • pp.21-33
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    • 1988
  • The purpose of this study was to investigate the effect of chlorambucil on the mouse Leydig cell by electron microscopy. Chlorambucil suspended in the 0.5N sodium bicarbonate(pH 8.0) was injected I.P.(intraperitoneal) at a dosage of level 20mg/kg for 1 weeks, 3 weeks and 5 weeks, respectively. The results obtained from this experiment are as follows: 1. One week after the administration of chlorambucil, there was an increase in heterochromatin, swelling and cristae disruption in some mitochondria, mild vacuolation between cells and the occurrence of membrane bound inclusions in some nuclei. 2. After 3 weeks, smooth endoplasmic reticulum dilations, cytoplasmic vacuolation, mitochondrial swelling, inner mitochondrial cristae disruption, membranous whorls, and intranuclear inclusions were observed in the treated cells. 3. After 5 weeks of treatment, most mitochondria were swollen and their membranes were severely disrupted. Further, smooth endoplasmic reticulum dilations and vacuolation of the cytoplasm were apparent in the treated Leydig cells. In addition numerous membranous whorls and intranuclear inclusion bodies were present. The nuclei displayed invaginatons of the nuclear membrane and large clumps of heterochromatin. From these results it is concluded the longer the duration of chlorambucil administration, the greater the degeneration of the nucleus and cytoplasmic organelles.

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Contributions of HO-1-Dependent MAPK to Regulating Intestinal Barrier Disruption

  • Zhang, Zhenling;Zhang, Qiuping;Li, Fang;Xin, Yi;Duan, Zhijun
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.175-183
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    • 2021
  • The mitogen-activated protein kinase (MAPK) pathway controls intestinal epithelial barrier permeability by regulating tight junctions (TJs) and epithelial cells damage. Heme oxygenase-1 (HO-1) and carbon monoxide (CO) protect the intestinal epithelial barrier function, but the molecular mechanism is not yet clarified. MAPK activation and barrier permeability were studied using monolayers of Caco-2 cells treated with tissue necrosis factor α (TNF-α) transfected with FUGW-HO-1 or pLKO.1-sh-HO-1 plasmid. Intestinal mucosal barrier permeability and MAPK activation were also investigated using carbon tetrachloride (CCl4) administration with CoPP (a HO-1 inducer), ZnPP (a HO-1 inhibitor), CO releasing molecule 2 (CORM-2), or inactived-CORM-2-treated wild-type mice and mice with HO-1 deficiency in intestinal epithelial cells. TNF-α increased epithelial TJ disruption and cleaved caspase-3 expression, induced ERK, p38, and JNK phosphorylation. In addition, HO-1 blocked TNF-α-induced increase in epithelial TJs disruption, cleaved caspase-3 expression, as well as ERK, p38, and JNK phosphorylation in an HO-1-dependent manner. CoPP and CORM-2 directly ameliorated intestinal mucosal injury, attenuated TJ disruption and cleaved caspase-3 expression, and inhibited epithelial ERK, p38, and JNK phosphorylation after chronic CCl4 injection. Conversely, ZnPP completely reversed these effects. Furthermore, mice with intestinal epithelial HO-1 deficient exhibited a robust increase in mucosal TJs disruption, cleaved caspase-3 expression, and MAPKs activation as compared to the control group mice. These data demonstrated that HO-1-dependent MAPK signaling inhibition preserves the intestinal mucosal barrier integrity by abrogating TJ dysregulation and epithelial cell damage. The differential targeting of gut HO-1-MAPK axis leads to improved intestinal disease therapy.

Effect of KGD1 Deletion on Cell Wall Biogenesis in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 KGD1 유전자 결손이 세포벽 생합성에 미치는 영향)

  • Kim, Sung-Woo;Ahn, Ki-Woong;Park, Yun-Hee;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.29-33
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    • 2010
  • KGD1 gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. We performed the gene disruption experiment to characterize the function of KGD1 gene, which encodes $\beta$-ketoglutarate dehydrogenase, in cell wall biosynthesis. The disruption of KGD1 showed the decreased growth rate, the increase of chitin synthases activity, alterations in cell wall composition, and increase of susceptibility to cell wall inhibitors such as Calcofluor white and Nikkomycin Z. These results suggested that KGD1 might be involved in cell wall biogenesis, especially the biosynthesis of $\beta$-1,6-glucan and chitin in S. cerevisaie.

The MAP Kinase Kinase Gene AbSte7 Regulates Multiple Aspects of Alternaria brassicicola Pathogenesis

  • Lu, Kai;Zhang, Min;Yang, Ran;Zhang, Min;Guo, Qinjun;Baek, Kwang-Hyun;Xu, Houjuan
    • The Plant Pathology Journal
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    • v.35 no.2
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    • pp.91-99
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    • 2019
  • Mitogen-activated protein kinase (MAPK) cascades in fungi are ubiquitously conserved signaling pathways that regulate stress responses, vegetative growth, pathogenicity, and many other developmental processes. Previously, we reported that the AbSte7 gene, which encodes a mitogen-activated protein kinase kinase (MAPKK) in Alternaria brassicicola, plays a central role in pathogenicity against host cabbage plants. In this research, we further characterized the role of AbSte7 in the pathogenicity of this fungus using ${\Delta}AbSte7$ mutants. Disruption of the AbSte7 gene of A. brassicicola reduced accumulation of metabolites toxic to the host plant in liquid culture media. The ${\Delta}AbSte7$ mutants could not efficiently detoxify cruciferous phytoalexin brassinin, possibly due to reduced expression of the brassinin hydrolase gene involved in detoxifying brassinin. Disruption of the AbSte7 gene also severely impaired fungal detoxification of reactive oxygen species. AbSte7 gene disruption reduced the enzymatic activity of cell walldegrading enzymes, including cellulase, ${\beta}$-glucosidase, pectin methylesterase, polymethyl-galacturonase, and polygalacturonic acid transeliminase, during host plant infection. Altogether, the data strongly suggest the MAPKK gene AbSte7 plays a pivotal role in A. brassicicola during host infection by regulating multiple steps, and thus increasing pathogenicity and inhibiting host defenses.

Effects of Sperm Membrane Disruption and Electrical Activation of Oocytes on In vitro Development and Transgenesis of Porcine Embryos Produced by Intracytoplasmic Sperm Injection

  • Shim, Sang Woo;Kim, Young Ha;Lee, Hoon Taek;Shim, Hosup
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.358-363
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    • 2008
  • The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

Disinfection of Water by Ultrasonic Irradiation (초음파 검사에 의한 수중의 살균처리)

  • 손종렬;유병성
    • Journal of environmental and Sanitary engineering
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    • v.14 no.1
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    • pp.54-61
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    • 1999
  • The industrial techniques of ultrasound have been used in the various fields, such as cleaning, medical surgery, emulsification, cell disruption etc. Especially the application of cell disruption was interested in the field of disinfection process in water by ultrasonic irradiation. It has been recognized that the ultrasounds are irradiated in aqueous solution, cavitation bubbles are generated and shock waves of high temperature and pressure are emitted as the bubbles are developed and finally broken, which function as a energy source to promote reaction efficiencies of various kinds of chemical reactions such as disinfection reaction in water. Therefore, this study was performed to apply the ultrasound for the disinfection method of infected drinking raw water and to discuss the limiting factors such as pH, sample volume and reaction temperature influenced on the removal efficiency of E. coli from experimental analysis of the results obtained in bench-scale plant. For the experiments to measure the influence of reaction parameters in the ultrasonic disinfection process, escalated reactivity of aqueous solutions was excellent when pH in aqueous solution was low, and sample volume was small. On the contrary, the reactivity of disinfection became elevated when reaction temperature was high. It was found that the rate constant of disinfection reaction was applied by Chick's law, reaction kinetics of Chick's law was irreversible and pseudo-first order at all the tested conditions.As a conclusion it appeared that the technology using ultrasonic irradiation can be applied to the treatment of disinfection in infected water which are difficult to be treated by conventional methods.

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Evaluation of COD Solubilization and Reduction of Waste Activated Sludge by pH Control (pH 조절을 통한 폐활성 슬러지의 COD 가용화 및 감량화 평가)

  • Kim, Youn Kwon;Moon, Yong Taik;Kim, Ji Yeon;Seo, In Seok
    • Journal of Korean Society of Water and Wastewater
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    • v.21 no.5
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    • pp.551-558
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    • 2007
  • From the view point of biological wastewater treatment, C/N ratio is one of the most important factor in biological nutrient removal process. However, municipal sewage in Korea is characterized by extremely low content of carbon source and relatively higher portion of N source. Accordingly, it is necessary to dose external carbon source in order to obtain higher degree of carbon source within the process. In this study, the effects of pH pretreatment as an alternative plan for increasing carbon source on the cell disruption and COD solubility of waste activated sludge were conducted under well defined experimental conditions. During 5 hours, the value of COD solubilization rate ($S_R$) at pH 11.5 is approximately 4.4 times higher than the value of $S_R$ at pH 9.5. It is expected that the level of SCOD increased due to the result from cell disruption. However, VSS/TSS ratio was not significantly changed after 5 hours. As Alkalinity changes gradually from less than 15, 30 and 60 meq NaOH/L, average RBCOD/SCOD fraction showed 34, 36 and 45%,respectively.

An Experimental Study on the Effect of Irradiation and cia- dichlorodiBmmineplatinum(II) on the myocardium of Rats (방사선조사와 cis-dichlorodismmineplstinum(II)가 휜쥐의 심근에 미치는 효과에 관한 실험적 연구)

  • Lee Kyung-Ja
    • Radiation Oncology Journal
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    • v.12 no.3
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    • pp.285-293
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    • 1994
  • Purpose : The study was designed to investigate the effect of cis-dichlorodiammineplatinum(II)(cis-DDP) on the radiation-induced cardiomyopathy in the rat. Materials and Methods : The myocardial damage was assessed by histopathologic changes. In radiation alone group, radiation dose ranged from 10-40 Gy X-ray in a single dose and in combined group, cis-dichlorodiammineplatinum(II) at a dose of 6 mg/kg was given intraperitoneally immediately after irradiation of same dose with X-ray alone group. Results : The early changes by radiation included congestion, inflammatory cell infiltrations and fibrosis in myocardial interstitium with focal myocardial necrosis, which was noted in 10 Gy group, Myocardial fibrosis was increased by increasing dose of radiation but myocardial necrosis was not Proportional to radiation dose. cis-DDP alone group showed minimal degeneration of myocardium with surrounded by inflammatory cell infiltrations. In combined group, myocardial fibrosis in 10 Gy group were similar to radiation alone group, but 30 Gy and 40 Gy groups showed severer changes. Electron microscopic examination showed disruption of Z-band and edema of mitochondria with decreased matrix density in 20 Gy radiation group which were severer in 40 Gy radiation group. Combined group showed endothelial changes and disruption of Z-band worse than radiation alone group as well as increased connective tissue, which was considered as a hallmark of late change in radiation-induced heart disease. Conclusion : This results showed minimal enhancement of the radiation-induced cardiomyopathy in rats by cis-DDP.

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Chlorella virus-mediated disruption of microalgal cell wall for biodiesel production (클로렐라 바이러스 매개 미세조류 세포벽 파쇄를 이용한 바이오 디젤 생산)

  • Kim, Soojin;Kim, Yeon-Soo
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.140-145
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    • 2018
  • The most energy-intensive processes in lipids extraction were the disruption of the cell wall of microalgae. Here, we tried to extract lipids through lysis using virus-infecting microalgae, to compare with those by the other two methods using microwave or ultrasonication. The lipids yield using viral infection was not significantly different from those using ultrasonication and microwave oven. This suggests that the same amount of lipids can be obtained with low energy and costs, as well as that microalgal lipids extraction by chlorella virus infection might provide the price competitiveness in biodiesel production even if it will be applied to mass production facilities.