• 제목/요약/키워드: Cycloheximide

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벼 종자에서 Xanthomonas campestris pv. oryzae의 분리를 위한 선택배지 (A Semiselective Medium for the Isolation of Xanthomonas campestris pv. oryzae from Rice Seed)

  • 김형무;송완엽;소인영;이두구
    • 한국식물병리학회지
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    • 제10권1호
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    • pp.13-17
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    • 1994
  • A semiselective agar medium (XCO) was developed for the isolation of bacterial blight pathogen, Xanthomonas campestris pv. oryzae, from rice seed. The medium contained yeast extract 1.0 g, peptone 2.0 g, sucrose 5.0 g, sodium glutamate 1.0 g, FeSO4.7H2O 0.05 g, Fe.EDTA 1 mg, cephalexin 20 mg, Evan blue (0.1%) 1.5 ml, bromcresol purple (0.1%) 2.5ml, cycloheximide 100 mg and agar 15.0 g per liter. Colonies of X. c. pv. oryzae were 4~5 mm in diameter, smooth, round, blue (darker center) and convex after 6 days incubation at 28$^{\circ}C$. The recovery of 6 isolates of X. c. pv. oryzae on the XOC medium ranged from 81% to 120% (mean 98.2%) in comparison to Wakimoto's medium. The number of saprophytic bacteria from 10 rice seed lots on XCO medium was reduced to 70.4% of that on Wakimoto's medium. The recovery of X. c. pv. oryzae added to rice seed on XOC medium ranged from 67% to 87% (mean 75.6%) of that on Wakimoto's medium.

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Bisifusarium Delphinoides, an Emerging Opportunistic Pathogen in a Burn Patient with Diabetes Mellitus

  • Park, Ji-Hyun;Oh, Junsang;Song, Ji-Sun;Kim, Jayoung;Sung, Gi-Ho
    • Mycobiology
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    • 제47권3호
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    • pp.340-345
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    • 2019
  • An 82-year-old man with diabetes was admitted to the emergency department with a third-degree burn on his left leg. The deep swab specimen from his left leg was cultured on Sabouraud dextrose agar without cycloheximide and incubated at $25^{\circ}C$ for 5 days. On the basis of morphological characteristics and multigene phylogenetic analyses of the internal transcribed spacer region of ribosomal DNA and partial fragments of beta-tubulin and translation elongation factor 1-alpha, the causal agent of fungal skin infection was identified as Bisifusarium delphinoides, which was newly introduced by accommodating a Fusarium dimerum species complex. Thus, we describe here the first case of skin infection caused by B. delphinoides on a burn patient with diabetes mellitus based on morphological observation and molecular analysis.

Characterization of Programmed Cell Death in the Silkworm Thoracic Ganglia during Postembryonic Periods

  • Kim, Soon-Ok;Kim, Mi-Young;Song, Hwa-Young;Kim, Jin-Hee;Kang, Pil-Don;Lee, Bong-Hee
    • Animal cells and systems
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    • 제11권1호
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    • pp.23-31
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    • 2007
  • Programmed cell death was characterized in the silkworm thoracic ganglia TG1, TG2 and TG3 during postembryonic periods by TUNEL assay. Apoptotic cells were detected in the three TGs of all larval stages except for day-1, 2 1st instar larvae, in which no apoptotic cells were found. From day-7 5th larva, the numbers of apoptotic cells were dramatically increased and peaked on day-1 pupa and day-2 pupa and then abruptly decreased. Apoptotic cells finally disappeared in day-1 adult. In-vivo injection of 20-hydroxyecdysone (20E) into day-8 5th larva resulted in a striking decrease of apoptotic cells. Actinomycin D (Act D) or cycloheximide (CHX), injected into hemolymph of day-8 5th larva, resulted in a decrease of apoptotic cells in the three TGs. Injection of caspase-8 and -3 inhibitors also blocked cellular apoptosis. These results will provide valuable information for understanding of cellular changes in the three TGs during metamorphosis of the insect species.

$Interferon-{\Upsilon}$ and Lipopolysaccaride Induce Mouse Guanylate-Binding Protein 3 (mGBP3) Expression in the Murine Macrophage Cell Line RAW264-7

  • Han, Byung-Hee
    • Archives of Pharmacal Research
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    • 제22권2호
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    • pp.130-136
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    • 1999
  • Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase which belongs to GTP-binding protein family. The present study showed that the expression of mGBP3 transcript was readily induced in a dose dependent fashion in the macrophage cell line RAW264.7 treated with either $interferon-{\gamma} (IFN-\gamma)$ or lipopolysaccaride (LPS). The expression of mGBP3 protein was also apparent by 4 and 6 h after the treatment of cells with IFN-\gamma (100 U/ml) or LPS ($1{\mu}g/ml$) , and remained at palteau for at least 24 h. Cycloheximide ($10{\mu}g/ml$) had no effect on the $IFN-\gamma-$ or LPS-induced mGBP3 expression, suggesting that the mGBP3 induction did not require further protein synthesis. Interestingly, a protein kinase C (PKC) inhibitor staurosporine (50 nM) abolished the induction of mGBP3 expression by LPS, but not by $IFN-{\gamma}$. These findings suggest that mGBP3 may be involved in the macrophage activation process and both IFN-\gamma and LS induce the mGBP3 expression through distinct signal transduction pathways.

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Pro-apoptotic Effects of S100A8 and S100A9 on human FIP1L1-PDGFRα+ Eosinophilic Leukemia Cells

  • Lee, Ji-Sook
    • 대한의생명과학회지
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    • 제27권2호
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    • pp.95-98
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    • 2021
  • The S100 family proteins act as inducers of cancer cell apoptosis and inflammatory mediators. This study examined the pro-apoptotic mechanism caused by S100A8 and S100A9 in human FIP1L1-PDGFRα-positive eosinophilic leukemia cells. S100A8 and S100A9 elicited the death of EoL-1 cells in a time and dose-dependent manner. The activation of PDGFRα was suppressed by a decrease in PDGFRα after treatment with S100A8 and S100A9. Cycloheximide, a translation inhibitor, suppressed PDGFRα expression from 1 h to 5 h, and a co-treatment with S100A8 and S100A9 boosted the decrease in expression. The phosphorylation and expression of STAT5 decreased after treatment with S100A8 and S100A9 in EoL-1 and imatinib-resistant (EoL-1-IR) cells. S100A8 and S100A9 induced the chemotaxis of EoL-1 cells but did not affect the chemoattraction of EoL-1-IR. These findings indicate the cell death mechanism due to S100 family proteins and the development of leukemia therapy using S100A8 and S100A9.

Stress Granules Inhibit Coxsackievirus B3-Mediated Cell Death via Reduction of Mitochondrial Reactive Oxygen Species and Viral Extracellular Release

  • Ji-Ye Park;Ok Sarah Shin
    • Journal of Microbiology and Biotechnology
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    • 제33권5호
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    • pp.582-590
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    • 2023
  • Stress granules (SGs) are cytoplasmic aggregates of RNA-protein complexes that form in response to various cellular stresses and are known to restrict viral access to host translational machinery. However, the underlying molecular mechanisms of SGs during viral infections require further exploration. In this study, we evaluated the effect of SG formation on cellular responses to coxsackievirus B3 (CVB3) infection. Sodium arsenite (AS)-mediated SG formation suppressed cell death induced by tumor necrosis factor-alpha (TNF-a)/cycloheximide (CHX) treatment in HeLa cells, during which G3BP1, an essential SG component, contributed to the modulation of apoptosis pathways. SG formation in response to AS treatment blocked CVB3-mediated cell death, possibly via the reduction of mitochondrial reactive oxygen species. Furthermore, we examined whether AS treatment would affect small extracellular vesicle (sEV) formation and secretion during CVB3 infection and modulate human monocytic cell (THP-1) response. CVB3-enriched sEVs isolated from HeLa cells were able to infect and replicate THP-1 cells without causing cytotoxicity. Interestingly, sEVs from AS-treated HeLa cells inhibited CVB3 replication in THP-1 cells. These findings suggest that SG formation during CVB3 infection modulates cellular response by inhibiting the release of CVB3-enriched sEVs.

Novel Detection Protocol for Erwinia amylovora in Orchard Soil after Removal of Infected Trees

  • Sujin Song;Byeori Kim;Kwang-Pyo Kim;Eunjung Roh
    • The Plant Pathology Journal
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    • 제40권3호
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    • pp.282-289
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    • 2024
  • Fire blight is a bacterial disease caused by Erwinia amylovora. In Korea, fire blight was first reported in 2015 in an orchard. If the infection is confirmed, all trees in the orchard must be removed and the orchard must remain closed for 3 years. Since 2020, if the number of trees infected with fire blight is less than 5% of the total trees in the orchard, only the infected tree and adjacent trees are removed in Korea. Three years after removal, the trees can be replanted after confirming that the orchard soil is free from E. amylovora. In this study, a protocol was established for detecting E. amylovora in soil via selective enrichment, using tryptic soy broth with 0.05% bile salts and 50 ㎍/ml cycloheximide, and real-time polymerase chain reaction. This protocol resulted in a 1,000-times improved detection limit for E. amylovora in soil samples compared to that in unenriched samples. Soil monitoring was performed for orchards where fire blight-infected trees had been removed 3-27 months prior; the selected orchards were monitored every 3 months. Monitoring confirmed that E. amylovora was not present in the soil at any site in any of the orchards. A new detection protocol facilitates the monitoring of E. amylovora in soil and could help permit the replanting of trees in orchards. Also monitoring results provide evidence that trees can be planted earlier.

무우 유식물의 자엽에서 Abscisic acid와 Polyamine 처리에 의한 아미노산 변화의 분석 (Analysis of the Change of Amino Acids by Abscisic acid and Polyamine Treatment in Radish Young Cotyledons)

  • 조봉희
    • 분석과학
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    • 제12권5호
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    • pp.441-446
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    • 1999
  • 무우 유식물의 자엽에서 Abscisic acid(ABA). ABA plus cyeloheximide(CH)와 ABA plus polyamine(PA) 처리로 유도되는 아미노산의 변화를 분석하였다. ABA 처리로 봄무우에서는 총 아미노산의 농도가 증가되었고, 겨울무우에서는 감소되었다. 봄무우에서는 친수성 아미노산인 proiine, glycine, serine과 cysteine 등이 증가되었고(특히 proline), 반면 겨울무우에서는 cysteine, leucine와 phenylalanine이 증가되었다(특히 cysteine과 leucine). CH는 ABA가 유도시킨 탈수에 대항할 수 있는 단백질의 합성을 방해하여 아미노산의 축적을 초래하였다. PA처리는 반대로 봄무우에서는 총 아미노산의 농도가 감소되었고, 겨울무우에서는 증가되었다. 이상의 결과는 축적된 친수성 아미노산과 단백질이 스트레스에 대항하여 탈수의 적응을 유도하였고, PA은 ABA에 의한 탈수 스트레스의 적응능력을 조절한다고 본다.

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Sperm Cytosolic Factor Activation for Bovine Somatic Cell Nuclear Transfer

  • Shin, Tae-Young
    • 한국수정란이식학회지
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    • 제26권3호
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    • pp.171-180
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    • 2011
  • In this study I report that in vitro development rates of bovine nuclear transfer embryos activated either with boar sperm cytosolic factor (SCF) or with ionomycin followed by cycloheximide (CHX) and subsequent in vivo developmental rates after embryo transfer are related to blastocyst quality as evaluated by apoptosis analysis. SCF was extracted from porcine semen then purified for post-activation injection after nuclear transfer. The optimal timing for SCF injection was determined to be at least 22 h post-IVM for parthenogenetic activation of bovine oocyies. A total of 364 oocytes were successfully enucleated and 268 (73.6%) fused and were injected with SCF. The survival rate of fused and injected embryos was 109/113 (96.5%) after 2 h. The cleavage rates of nuclear transfer embryos after 3 d of culture in the ionomycin/CHX treated group were significantly higher than those of the SCF-activated group (93.3% vs 81.7%, p<0.01, respectively). However, at 7 d and 9 d there was no significant difference between the total developmental rates to blastocyst for either treatment group. Total blastocyst cell numbers were also not significantly different between the two activation treatments (ionomycin/CHX: 149.5${\pm}$7.7 vs. SCF: 139.3${\pm}$4.4 cells). In contrast, the apoptotic levels in the SCF blastocysts were higher than those produced after the chemical treatment (28.2${\pm}$5.1% vs. 8.8${\pm}$0.6%, respectively). A total of 18 expanded or hatching blastocysts was transferred to nine synchronized recipients in each activation group; 5/9 (55.5%) and 2/9 (22.2%) were pregnant at 40 d in the ionomycin/CHX treatment and SCF activated group, respectively. However, only one went to term in the ionomycin/CHX treatment while none of the pregnancies from the SCF group were maintained by 90 d. In conclusion, these results suggest that SCF derived from different species is a limited activator to be used for activation after bovine nuclear transfer in lieu of a chemical activation protocol.