• 제목/요약/키워드: effector analysis

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Immunohistochemical Analysis of Phosphorylated Akt Protein Expression in Gastric Carcinomas (위암의 Phosphorylated Akt 단백질의 발현)

  • Lee Sug Hyung;Lee Jong Woo;Park Won Sang;Lee Jung Young;Yoo Nam Jin;Kim Su Young
    • Journal of Gastric Cancer
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    • v.3 no.2
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    • pp.88-92
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    • 2003
  • Purpose: Mounting evidence suggests that alterations of Akt/protein kinase B (PKB) play an important role in tumorigenesis. Phosphorylated Akt regulates many of the key effector molecules involved in apoptosis, angiogenesis, and cell-cycle progression during tumorigenesis. The expression of phosphorylated Akt has been described in some human malignancies, but not in primary human gastric cancer. The purpose of this study was to explore the expression status of phosphorylated Akt protein in gastric carcinomas. Materials and Methods: In the current study, we analyzed the expression of phosphorylated Akt protein in 60 advanced gastric adenocarcinomas by using immunohistochemistry and a tissue microarray approach. Results: Immunopositivity (defined as $\geq\30\%$) was observed for the phosphorylated Akt in 42 ($70\%$) of the 60 cancers. Normal gastric mucosal cells showed no or weak expression of phosphorylated Akt protein. Conclusion: Taken together, these results indicate that Akt is frequently activated in gastric adenocarcinoma cells and suggest that phosphorylayed Akt may play a role in the development of human gastric adenocarcinomas.

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Homologous Expression and T3SS-Dependent Secretion of TAP-Tagged Xo2276 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract and Its Direct In Vitro Recognition of Putative Target DNA Sequence

  • Kim, Seunghwan;Nguyen, Thi-Dieu-Hanh;Lee, Joohee;Hong, Myoung-Ki;Pham, Tan-Viet;Ahn, Yeh-Jin;Lee, Byoung-Moo;Han, Ye Sun;Kim, Dong-Eun;Kim, Jeong-Gu;Kang, Lin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.22-28
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    • 2013
  • Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoosusceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAPtagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.

Pi5 and Pii Paired NLRs Are Functionally Exchangeable and Confer Similar Disease Resistance Specificity

  • Vo, Kieu Thi Xuan;Lee, Sang-Kyu;Halane, Morgan K.;Song, Min-Young;Hoang, Trung Viet;Kim, Chi-Yeol;Park, Sook-Young;Jeon, Junhyun;Kim, Sun Tae;Sohn, Kee Hoon;Jeon, Jong-Seong
    • Molecules and Cells
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    • v.42 no.9
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    • pp.637-645
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    • 2019
  • Effector-triggered immunity (ETI) is an effective layer of plant defense initiated upon recognition of avirulence (Avr) effectors from pathogens by cognate plant disease resistance (R) proteins. In rice, a large number of R genes have been characterized from various cultivars and have greatly contributed to breeding programs to improve resistance against the rice blast pathogen Magnaporthe oryzae. The extreme diversity of R gene repertoires is thought to be a result of co-evolutionary history between rice and its pathogens including M. oryzae. Here we show that Pii is an allele of Pi5 by DNA sequence characterization and complementation analysis. Pii-1 and Pii-2 cDNAs were cloned by reverse transcription polymerase chain reaction from the Pii-carrying cultivar Fujisaka5. The complementation test in susceptible rice cultivar Dongjin demonstrated that the rice blast resistance mediated by Pii, similar to Pi5, requires the presence of two nucleotide-binding leucine-rich repeat genes, Pii-1 and Pii-2. Consistent with our hypothesis that Pi5 and Pii are functionally indistinguishable, the replacement of Pii-1 by Pi5-1 and Pii-2 by Pi5-2, respectively, does not change the level of disease resistance to M. oryzae carrying AVR-Pii. Surprisingly, Exo70F3, required for Pii-mediated resistance, is dispensable for Pi5-mediated resistance. Based on our results, despite similarities observed between Pi5 and Pii, we hypothesize that Pi5 and Pii pairs require partially distinct mechanisms to function.

Effect of immune-enhancing enteral nutrition formula enriched with plant-derived n-3 fatty acids on natural killer cell activity in rehabilitation patients

  • Cho, Jung Min;Choi, Hyo Seon;Cho, Youn Soo;Park, So Young;Kim, Deog Young;Lee, Jong Ho
    • Nutrition Research and Practice
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    • v.13 no.5
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    • pp.384-392
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    • 2019
  • BACKGROUND/OBJECTIVES: Enteral nutrition formulas with immune-enhancing nutrients, such as n-3 fatty acids, may manage patients' nutritional status and pathophysiological processes. The aim of our study was to investigate natural killer (NK) cell activity alterations and related cytokine changes resulting from feeding with soybean oil-containing enteral nutrition formula (control group) and plant-derived n-3 fatty acid-enriched enteral nutrition formula. SUBJECTS/METHODS: Subjects participated for 14 consecutive days and consumed enteral formula containing canola and flaxseed oil (n3EN, test group) in nonsurgical patients hospitalized for rehabilitation. Blood samples were collected on the first day and 14 days after the consumption of each formula daily, and anthropometric parameters were collected. Hematology and biochemical values were analyzed, and NK cell activities and serum cytokine concentration were measured. A total of sixty subjects were included in the analysis, excluding dropouts. RESULTS: No significant differences were found in biochemical parameters. The n3EN group's NK cell activities at effector:tumor cell ratios of 10:1, 5:1, 2.5:1 and 0.625:1 were significantly higher than those of the control group after two weeks (P < 0.05). However, there were no statistically significant differences in serum cytokine interleukin (IL)-12, $interferon-{\gamma}$, $IL-1{\beta}$, IL-6 and tumor necrosis $factor-{\alpha}$ values between the two groups. CONCLUSIONS: In conclusion, this study elucidates the beneficial effects of plant-derived n-3 fatty acid supplementation in enteral formula on NK cell activity.

Discrimination and bifurcation analysis of tumor immune interaction in fractional form

  • Taj, Muhammad;Khadimallah, Mohamed A.;Hussain, Muzamal;Rashid, Yahya;Ishaque, Waqas;Mahmoud, S.R.;Din, Qamar;Alwabli, Afaf S.;Tounsi, Abdelouahed
    • Advances in nano research
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    • v.10 no.4
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    • pp.359-371
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    • 2021
  • A tumor immune interaction is a main topic of interest in the last couple of decades because majority of human population suffered by tumor, formed by the abnormal growth of cells and is continuously interacted with the immune system. Because of its wide range of applications, many researchers have modeled this tumor immune interaction in the form of ordinary, delay and fractional order differential equations as the majority of biological models have a long range temporal memory. So in the present work, tumor immune interaction in fractional form provides an excellent tool for the description of memory and hereditary properties of inter and intra cells. So the interaction between effector-cells, tumor cells and interleukin-2 (IL-2) are modeled by using the definition of Caputo fractional order derivative that provides the system with long-time memory and gives extra degree of freedom. Moreover, in order to achieve more efficient computational results of fractional-order system, a discretization process is performed to obtain its discrete counterpart. Furthermore, existence and local stability of fixed points are investigated for discrete model. Moreover, it is proved that two types of bifurcations such as Neimark-Sacker and flip bifurcations are studied. Finally, numerical examples are presented to support our analytical results.

Effect of Water Extract of Sparasis crispa on the Expression of TNF-α, iNOS and IL-1β Genes in RAW 264.7 Cells (꽃송이버섯 추출물이 RAW 264.7 세포에서 TNF-α, iNOS, IL-1β 유전자 발현에 미치는 영향)

  • Han, Hyo-Sang
    • Journal of The Korean Society of Integrative Medicine
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    • v.9 no.1
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    • pp.163-171
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    • 2021
  • Purpose : The purpose of this study was to examine the anti-inflammatory effects of Sparassis crispa (SC). SC is a well-known traditional herbal remedy and its mushroom is used for treatment of inflammation. Many diseases that are increasing recently have characteristics of inflammatory diseases. Researchers are finding bioactive substances from natural products that can promote treatment and prevention of inflammation. We investigated the effect of water extracted from SC on the expression of effector genes involved in the function of RAW 264.7 cells. Methods : Effects of RAW 264.7 cells on cell viability, antioxidation, and mRNA expression were examined using water extracts from SC. A 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to determine the effect of water extracts from SC on cell viability in RAW 264.7 cells. Inflammation of RAW 264.7 cells induced by lipopolysaccharide (LPS) treatment and expression levels of inflammatory cytokine TNF-α, iNOS and IL-1β gene were analyzed using quantitative reverse transcription PCR (qRT-PCR) analysis. Results : The MTS assay was performed on RAW 264.7 cells after treatment with various concentrations of water extracts of SC. Treatment of RAW 264.7 cells with water extracts from SC and LPS at a concentration of 0.125, 0.5 mg/㎖ for twenty four hours promoted mRNA expression of TNF-α, iNOS and IL-1β. Conclusion : MTS assay was applied to RAW 264.7 cells after various concentrations of water extracts of SC. Through experimental demonstration of anti-oxidant and anti-inflammatory effects of water extracts from SC, we suggest that SC is a valuable material for the prevention and treatment of various inflammatory diseases.

Digital Filter Model for Analog Helical Coil Spring Reverberator (헬리컬 코일 스프링 잔향기의 디지털 필터 모델)

  • Park Joon;Chon Sang-Bae;Lee Jong-Hoon;Sung Koeng-Mo;Song Sang-Seob
    • The Journal of the Acoustical Society of Korea
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    • v.25 no.6
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    • pp.291-297
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    • 2006
  • This paper proposes a new Digital Reverberator that models Analog Helical Coil Spring Reverberator for guitar amplifiers. While the conventional digital reverberators are proposed to provide better sound field mainly based on room acoustics, no algorithm or analysis of digital reverberators those model Helical Coil Spring Reverberator was proposed. Considering the fact that approximately $70{\sim}80$ percent of guitar amplifiers are still with Helical Coil Spring Reverberator, research was performed based not on Room Acoustics but on Helical Coil Spring Reverberator itself as an effector. After performing simulations with proposed algorithm, it was confirmed that the Digital Reverberator by proposed algorithm provides perceptually equivalent response to the conventional Analog Helical Coil Spring Reverberators.

Bioactivity-Guided Fraction from Viscera of Abalone, Haliotis discus hannai Suppresses Cellular Basophils Activation and Anaphylaxis in Mice

  • Kap Seong Choi;Tai-Sun Shin;Ginnae Ahn;Shin Hye Kim;Jiyeon Chun;Mina Lee;Dae Heon Kim;Han-Gil Choi;Kyung-Dong Lee;Sun-Yup Shim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.379-386
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    • 2024
  • Basophils and mast cells are specialized effector cells in allergic reactions. Haliotis discus hannai (abalone), is valuable seafood. Abalone male viscera, which has a brownish color and has not been previously reported to show anti-allergic activities, was extracted with acetone. Six different acetone/hexane fractions (0, 10, 20, 30, 40, and 100%) were obtained using a silica column via β-hexosaminidase release inhibitory activity-guided selection in phorbol myristate acetate and a calcium ionophore, A23187 (PMACI)-induced human basophils, KU812F cells. The 40% acetone/hexane fraction (A40) exhibited the strongest inhibition of PMACI-induced-β-hexosaminidase release. This fraction dose-dependently inhibited reactive oxygen species (ROS) production and calcium mobilization without cytotoxicity. Western blot analysis revealed that A40 down-regulated PMACI-induced MAPK (ERK 1/2, p-38, and JNK) phosphorylation, and the NF-κB translocation from the cytosol to membrane. Moreover, A40 inhibited PMACI-induced interleukin (IL)-1β, IL-6, and IL-8 production. Anti-allergic activities of A40 were confirmed based on inhibitory effects on IL-4 and tumor necrosis factor alpha (TNF-α) production in compound (com) 48/80-induced rat basophilic leukemia (RBL)-2H3 cells. A40 inhibited β-hexosaminidase release and cytokine production such as IL-4 and TNF-α produced by com 48/80-stimulated RBL-2H3 cells. Furthermore, it's fraction attenuated the IgE/DNP-induced passive cutaneous anaphylaxis (PCA) reaction in the ears of BALB/c mice. Our results suggest that abalone contains the active fraction, A40 is a potent therapeutic and functional material to treat allergic diseases.

Comparative Genomic Analysis of Pathogenic Factors of Pectobacterium Species Isolated in South Korea Using Whole-Genome Sequencing

  • Jee, Samnyu;Kang, In-Jeong;Bak, Gyeryeong;Kang, Sera;Lee, Jeongtae;Heu, Sunggi;Hwang, Ingyu
    • The Plant Pathology Journal
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    • v.38 no.1
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    • pp.12-24
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    • 2022
  • In this study, we conducted whole-genome sequencing with six species of Pectobacterium composed of seven strains, JR1.1, BP201601.1, JK2.1, HNP201719, MYP201603, PZ1, and HC, for the analysis of pathogenic factors associated with the genome of Pectobacterium. The genome sizes ranged from 4,724,337 bp to 5,208,618 bp, with the GC content ranging from 50.4% to 52.3%. The average nucleotide identity was 98% among the two Pectobacterium species and ranged from 88% to 96% among the remaining six species. A similar distribution was observed in the carbohydrate-active enzymes (CAZymes) class and extracellular plant cell wall degrading enzymes (PCWDEs). HC showed the highest number of enzymes in CAZymes and the lowest number in the extracellular PCWDEs. Six strains showed four subsets, and HC demonstrated three subsets, except hasDEF, in type I secretion system, while the type II secretion system of the seven strains was conserved. Components of human pathogens, such as Salmonella pathogenicity island 1 type type III secretion system (T3SS) and effectors, were identified in PZ1; T3SSa was not identified in HC. Two putative effectors, including hrpK, were identified in seven strains along with dspEF. We also identified 13 structural genes, six regulator genes, and five accessory genes in the type VI secretion system (T6SS) gene cluster of six Pectobacterium species, along with the loss of T6SS in PZ1. HC had two subsets, and JK2.1 had three subsets of T6SS. With the GxSxG motif, the phospholipase A gene did locate among tssID and duf4123 genes in the T6SSa cluster of all strains. Important domains were identified in the vgrG/paar islands, including duf4123, duf2235, vrr-nuc, and duf3396.

MEKK3 and Survivin Expression in Cervical Cancer: Association with Clinicopathological Factors and Prognosis

  • Cao, Xue-Quan;Lu, Hong-Sheng;Zhang, Ling;Chen, Li-Li;Gan, Mei-Fu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5271-5276
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    • 2014
  • Mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 3 (MEKK3) is an important protein kinase and a member of the MAPK family, which regulates cellular responses to environmental stress and serves as key integration points along the signal transduction cascade that not only link diverse extracellular stimuli to subsequent signaling molecules but also amplify the initiating signals to ultimately activate effector molecules and induce cell proliferation, differentiation and survival. To explore the relationship between MEKK3 and cell apoptosis, clinicopathology and prognosis, we characterize the expression of MEKK3 and survivin in cervical cancer. MEKK3 and survivin expression was measured by RT-PCR and Western blotting of fresh surgical resections from 30 cases of cervical cancer and 25 cases of chronic cervicitis. Protein expression was detected by tissue microarray and immunochemistry (En Vision) in 107 cases of cervical cancer, 86 cases of cervical intraepithelial neoplasia (CIN), and 35 cases of chronic cervicitis. Expression patterns were analyzed for their association with clinicopathological factors and prognosis in cervical cancer. Expression of MEKK3 and survivin mRNA was significantly higher in cervical cancer than in the controls (p<0.05). MEKK3 and survivin expression differed significantly between cervical carcinoma, CIN, and cervicitis (p<0.05) and correlated with clinical stage, infiltration depth, and lymph node metastasis (p<0.05). MEKK3 expression was positively correlated with survivin (p<0.05). Kaplan-Meier survival analysis showed that MEKK3 and survivin expression, lymph node metastasis, depth of invasion, and FIGO stage reduce cumulative survival. Cox multivariate regression analysis showed that MEKK3, survivin, and clinical staging are independent prognostic factors in cervical cancer (p<0.05). Expression of MEKK3 and survivin are significantly increased in cervical cancer, their overexpression participating in the occurrence and development of cervical cancer, with protein expression and clinical staging acting as independent prognostic factors for patients with cervical cancer.