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HeLa Cells Containing a Truncated Form of DNA Polymerase Beta are More Sensitized to Alkylating Agents than to Agents Inducing Oxidative Stress

  • Khanra, Kalyani;Chakraborty, Anindita;Bhattacharyya, Nandan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8177-8186
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    • 2016
  • The present study was aimed at determining the effects of alkylating and oxidative stress inducing agents on a newly identified variant of DNA polymerase beta ($pol{\beta}{\Delta}_{208-304}$) specific for ovarian cancer. $Pol{\beta}{\Delta}_{208-304}$ has a deletion of exons 11-13 which lie in the catalytic part of enzyme. We compared the effect of these chemicals on HeLa cells and HeLa cells stably transfected with this variant cloned into in pcDNAI/neo vector by MTT, colony forming and apoptosis assays. $Pol{\beta}{\Delta}_{208-304}$ cells exhibited greater sensitivity to an alkylating agent and less sensitivity towards $H_2O_2$ and UV when compared with HeLa cells alone. It has been shown that cell death in $Pol{\beta}{\Delta}_{208-304}$ transfected HeLa cells is mediated by the caspase 9 cascade. Exon 11 has nucleotidyl selection activity, while exons 12 and 13 have dNTP selection activity. Hence deletion of this part may affect polymerizing activity although single strand binding and double strand binding activity may remain same. The lack of this part may adversely affect catalytic activity of DNA polymerase beta so that the variant may act as a dominant negative mutant. This would represent clinical significance if translated into a clinical setting because resistance to radiation or chemotherapy during the relapse of the disease could be potentially overcome by this approach.

A PAS-Containing Histidine Kinase is Required for Conidiation, Appressorium Formation, and Disease Development in the Rice Blast Fungus, Magnaporthe oryzae

  • Shin, Jong-Hwan;Gumilang, Adiyantara;Kim, Moon-Jong;Han, Joon-Hee;Kim, Kyoung Su
    • Mycobiology
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    • v.47 no.4
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    • pp.473-482
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    • 2019
  • Rice blast disease, caused by the ascomycete fungus Magnaporthe oryzae, is one of the most important diseases in rice production. PAS (period circadian protein, aryl hydrocarbon receptor nuclear translocator protein, single-minded protein) domains are known to be involved in signal transduction pathways, but their functional roles have not been well studied in fungi. In this study, targeted gene deletion was carried out to investigate the functional roles of the PAS-containing gene MoPAS1 (MGG_02665) in M. oryzae. The deletion mutant ΔMopas1 exhibited easily wettable mycelia, reduced conidiation, and defects in appressorium formation and disease development compared to the wild type and complemented transformant. Exogenous cAMP restored appressorium formation in ΔMopas1, but the shape of the restored appressorium was irregular, indicating that MoPAS1 is involved in sensing the hydrophobic surface. To examine the expression and localization of MoPAS1 in M. oryzae during appressorium development and plant infection, we constructed a MoPAS1:GFP fusion construct. MoPAS1:GFP was observed in conidia and germ tubes at 0 and 2 h post-infection (hpi) on hydrophobic cover slips. By 8 hpi, most of the GFP signal was observed in the appressoria. During invasive growth in host cells, MoPAS1:GFP was found to be fully expressed in not only the appressoria but also invasive hyphae, suggesting that MoPAS may contribute to disease development in host cells. These results expand our knowledge of the roles of PAS-containing regulatory genes in the plant-pathogenic fungus M. oryzae.

Comparison of Remaining Data According to Deletion Events on Microsoft SQL Server (Microsoft SQL Server 삭제 이벤트의 데이터 잔존 비교)

  • Shin, Jiho
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.27 no.2
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    • pp.223-232
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    • 2017
  • Previous research on data recovery in Microsoft SQL Server has focused on restoring data based on in the transaction log that might have deleted records exist. However, there was a limit that was not applicable if the related transaction log did not exist or the physical database file was not connected to Server. Since the suspect in the crime scene may delete the data records using a different deletion statements besides "delete", we need to check the remaining data and a recovery possibility of the deleted record. In this paper, we examined the changes "Page Allocation information" of the table, "Unallocation deleted data", "Row Offset Array" in the page according to "delete", "truncate" and "drop" events. Finally it confirmed the possibility of data recovery and availability of management tools in Microsoft SQL Server digital forensic investigation.

The Magas1 Gene is Involved in Pathogenesis by Affecting Penetration in Metarhizium acridum

  • Cao, Yueqing;Zhu, Xiangxian;Jiao, Run;Xia, Yuxian
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.889-893
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    • 2012
  • Appressorium is a specialized infection structure of filamentous pathogenic fungi and plays an important role in establishing a pathogenic relationship with the host. The Egh16/Egh16H family members are involved in appressorium formation and pathogenesis in pathogenic filamentous fungi. In this study, a homolog of Egh16H, Magas1, was identified from an entomopathogenic fungus, Metarhizium acridum. The Magas1 protein shared a number of conserved motifs with other Egh16/Egh16H family members and specifically expressed during the appressorium development period. Magas1-EGFP fusion expression showed that Magas1 protein was not localized inside the cell. Deletion of the Magas1 gene had no impact on vegetative growth, conidiation and appressorium formation, but resulted in a decreased mortality of host insect when topically inoculated. However, the mortality was not significant between the Magas1 deletion mutant and wild-type treatment when the cuticle was bypassed by injecting conidia directly into the hemocoel. Our results suggested that Magas1 may influence virulence by affecting the penetration of the insects' cuticle.

Comparisons of growth and biofilm production with Vibrio fluvialis and mutants deficient in oligopeptide permease gene (Vibrio fluvialis의 Oligopeptide Permease Gene 결손에 의한 생육과 Biofilm 생산의 비교)

  • Lee, Eun Mi;Ahn, Sun Hee;Kong, In Soo
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.84-90
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    • 2006
  • Various ${\Delta}opp$ mutants of Vibrio fluvialis were constructed by allelic exchange method. The mutants occurred in target genes were confirmed by PCR and Southern hybridization analyses. After the exact mutants were identified, cell growth and biofilm production were examined using the respective mutants. The growth of wild strain was more rapid than mutants within 4hr incubation. Thereafter, the growth of wild strain and mutants reached to same level. When the productivities of wild strain and mutants were examined, ${\Delta}oppA$ mutant showed the highest productivity. Though ${\Delta}oppC,D$ and F mutants produced the lower production than that of ${\Delta}oppA$ mutant, the productivities of those mutants were much higher than that of wild strain.

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Influences of Hinge Region of a Systhetic Antimicrobial Peptide, Cecropin A(1-13)-Melittin(1-13) Hybrid on Antibiotic Activity

  • 신송엽;강주현;이동건;장소윤;서무열;김길룡;함경수
    • Bulletin of the Korean Chemical Society
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    • v.20 no.9
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    • pp.1078-1084
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    • 1999
  • A synthetic cecropin A(1-13)-melittin(1-13) [CA-ME] hybrid peptide was known to be an antimicrobial peptide having strong antibacterial, antifungal and antitumor activity with minimal cytotoxic effect against human erythrocyte. Analogues were synthesized to investigate the influences of the flexible hinge region of CA-ME on the antibiotic activity. Antibiotic activity of the peptides was measured by the growth inhibition against bac-terial, fungal and tumor cells and vesicle-aggregating or disrupting activity. The deletion of Gln-Gly-Ile (P1) or Gly-Gln-Gly-Ile-Gly (P3) from CA-ME brought about a significant decrease on the antibiotic activities. In contrast, Gly-Ile-Gly deletion (P2) from CA-ME or Pro insertion (P5) instead of Gly-Gln-Gly-Ile-Gly of CA-ME retained antibiotic activity. This result indicated that the flexible hinge or β-bend structure provided by Gly-Gln-Gly-Ile-Gly, Gln-Gly, or Pro in the central region of the peptides is requisite for its effective antibiotic activity and may facilitate easily the hydrophobic C-terminal region of the peptide to penetrate the lipid bilayers of the target cell membrane. In contrast, P4 and P6 with Gly-Gln-Gly-Pro-Gly or Gly-Gln-Pro in the central region of the peptide caused a drastic reduction on the antibiotic activities. This result suggested that the con-secutive β-bend structure provided by Gly-Gln-Gly-Pro-Gly or Gly-Gln-Pro in the central hinge region of the peptide seems to interrupt the ion channel/pore formation on the target cell membranes.

Genetic overgrowth syndrome: A single center's experience

  • Cheon, Chong Kun;Kim, Yoo-Mi;Yoon, Ju Young;Kim, Young A
    • Journal of Genetic Medicine
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    • v.15 no.2
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    • pp.64-71
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    • 2018
  • Purpose: Overgrowth syndromes are conditions that involve generalized or localized areas of excess growth. In this study, the clinical, molecular, and genetic characteristics of Korean patients with overgrowth syndrome were analyzed. Materials and Methods: We recruited 13 patients who presented with overgrowth syndrome. All patients fulfilled inclusion criteria of overgrowth syndrome. Analysis of the clinical and molecular investigations of patients with overgrowth syndrome was performed retrospectively. Results: Among the 13 patients with overgrowth syndrome, 9 patients (69.2%) were found to have molecular and genetic causes. Among the seven patients with Sotos syndrome (SS), two had a 5q35microdeletion that was confirmed by fluorescent in situ hybridization. In two patients with SS, intragenic mutations including a novel mutation, c.5993T>A (p.M1998L), were found by Sanger sequencing. One patient had one copy deletion of NDS1 gene which was confirmed by multiplex ligation-dependent probe amplification. Among five patients with Beckwith-Wiedemann syndrome, three had aberrant imprinting control regions; 2 hypermethylation of the differentially methylated region of H19, 1 hypomethylation of the differentially methylated region of Kv. In one patient displaying overlapping clinical features of SS, a de novo heterozygous deletion in the chromosomal region 7q22.1-22.3 was found by single nucleotide polymorphism-based microarray. Conclusion: Considering high detection rate of molecular and genetic abnormalities in this study, rigorous investigations of overgrowth syndrome may be an important tool for the early diagnosis and genetic counseling. A detailed molecular analysis of the rearranged regions may supply the clues for the identification of genes involved in growth regulation.

Forebrain glutamatergic neuron-specific Ctcf deletion induces reactive microgliosis and astrogliosis with neuronal loss in adult mouse hippocampus

  • Kwak, Ji-Hye;Lee, Kyungmin
    • BMB Reports
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    • v.54 no.6
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    • pp.317-322
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    • 2021
  • CCCTC-binding factor (CTCF), a zinc finger protein, is a transcription factor and regulator of chromatin structure. Forebrain excitatory neuron-specific CTCF deficiency contributes to inflammation via enhanced transcription of inflammation-related genes in the cortex and hippocampus. However, little is known about the long-term effect of CTCF deficiency on postnatal neurons, astrocytes, or microglia in the hippocampus of adult mice. To address this, we knocked out the Ctcf gene in forebrain glutamatergic neurons (Ctcf cKO) by crossing Ctcf-floxed mice with Camk2a-Cre mice and examined the hippocampi of 7.5-10-month-old male mice using immunofluorescence microscopy. We found obvious neuronal cell death and reactive gliosis in the hippocampal cornu ammonis (CA)1 in 7.5-10-month-old cKO mice. Prominent rod-shaped microglia that participate in immune surveillance were observed in the stratum pyramidale and radiatum layer, indicating a potential increase in inflammatory mediators released by hippocampal neurons. Although neuronal loss was not observed in CA3, and dentate gyrus (DG) CTCF depletion induced a significant increase in the number of microglia in the stratum oriens of CA3 and reactive microgliosis and astrogliosis in the molecular layer and hilus of the DG in 7.5-10-month-old cKO mice. These results suggest that long-term Ctcf deletion from forebrain excitatory neurons may contribute to reactive gliosis induced by neuronal damage and consequent neuronal loss in the hippocampal CA1, DG, and CA3 in sequence over 7 months of age.

Comparative Evaluation of Data Processing Performance between MySQL and Redis (MySQL과 Redis의 데이터 처리 성능 비교 평가)

  • Hyeok Bang;Seo-Hyeon Kim;Sanghoon Jeon
    • Journal of Internet Computing and Services
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    • v.25 no.3
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    • pp.35-41
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    • 2024
  • As online activities have rapidly increased due to recent digital changes and the impact of COVID-19, the importance of large-scale data processing and maintenance is increasing. This study compares the performance of the two main types of databases widely used for data storage and management: Relational Database Management Systems (RDBMS) and Non-Relational Databases (NoSQL). Specifically, we measured and evaluated the execution time of data insertion, query, and deletion functions using MySQL, a representative example of RDBMS, and Redis, a representative example of NoSQL. The experimental results showed that Redis showed performance about 5.84 times faster in data insertion, 6.61 times faster in query, and 12.33 times faster in deletion than MySQL. These results demonstrate that Redis provides superior performance, especially in environments requiring large-scale data processing and maintenance. Therefore, companies and online service providers can choose NoSQL databases such as Redis to ensure more efficient data management solutions. We hope this study will be an essential reference when selecting a database based on data processing performance.

Function of Dual Specificity Kinase, ScKns1, in Adhesive and Filamentous Growth of Saccharomyces cerevisiae (Saccharomyces cerevisiae의 균사형 생장에서 이중 특이성 인산화 효소, ScKns1p의 기능 분석)

  • Park, Yun-Hee;Yang, Ji-Min;Yang, So-Young;Kim, Sang-Mi;Cho, Young-Mi;Park, Hee-Moon
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.110-116
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    • 2011
  • In the previous study with the Saccharomyces cerevisiae S288c strains, no known function of the dual specificity kinase, ScKns1, was reported because its gene deletion did not show any noticeable phenotypic changes. Recent study with fission yeast, however, revealed the involvement of the LAMMER kinase in flocculation, filamentous growth, oxidative stress, and so on. Therefore we made Sckns1-deletion mutants with the ${\Sigma}1278b$-background, with which one can induce filamentous and adhesive growth in contrast to those of the S288c-background. The $Sckns1{\Delta}$ strains of both haploid and diploid showed defect in filamentous growth under conditions for inducing the filamentous growth such as nitrogen starvation and butanol treatment. Both kinds of the deletion mutants also showed decrease in adhesive growth on agar surface. Interestingly enough the defects of the $Sckns1{\Delta}$ strains were suppressed by the over-expression of each gene for the components of the MAPK signaling pathway such as STE11, STE12, and TEC1, respectively, but not by the upstream components, RAS2 and STE20, respectively. Although further investigations are required, these results indicate that the ScKns1 may act in place between the Ste20 and the Ste11 of the S. cerevisiae MAPK cascade.