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Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

Let-7c Inhibits NSCLC Cell Proliferation by Targeting HOXA1

  • Zhan, Min;Qu, Qiang;Wang, Guo;Liu, Ying-Zi;Tan, Sheng-Lan;Lou, Xiao-Ya;Yu, Jing;Zhou, Hong-Hao
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.387-392
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    • 2013
  • Objective: The aim of the present study was to explore mechanisms by which let-7c suppresses NSCLC cell proliferation. Methods: The expression level of let-7c was quantified by qRT-PCR. A549 and H1299 cells were transfected with let-7c mimics to restore the expression of let-7c. The effects of let-7c were then assessed by cell proliferation, colony formation and cell cycle assay. Mouse experiments were used to confirm the effect of let-7c on tumorigenicity in vivo. Luciferase reporter assays and Western blotting were performed to identify target genes for let-7c. Results: HOXA1 was identified as a novel target of let-7c. MTS, colony formation and flow cytometry assays demonstrated that forced expression of let-7c inhibited NSCLC cell proliferation by inducing G1 arrest in vitro, consistent with inhibitory effects induced by knockdown of HOXA1. Mouse experiments demonstrated that let-7c expression suppressed tumorigenesis. Furthermore, we found that let-7c could regulate the expression of HOXA1 downstream effectors CCND1, CDC25A and CDK2. Conclusions: Collectively, these results demonstrate let-7c inhibits NSCLC cell proliferation and tumorigenesis by partial direct targeting of the HOXA1 pathway, which suggests that restoration of let-7c expression may thus offer a potential therapeutic intervention strategy for NSCLC.

Survival Association and Cell Cycle Effects of B7H3 in Neuroblastoma

  • Zhang, Haibo;Zhang, Jinsen;Li, Chunjie;Xu, Hao;Dong, Rui;Chen, Clark C.;Hua, Wei
    • Journal of Korean Neurosurgical Society
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    • v.63 no.6
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    • pp.707-716
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    • 2020
  • Objective : The function of B7H3, a member of the B7 family of proteins, in neuroblastoma (NB) remains poorly characterized. Here we examine the expression pattern of B7H3 in clinical NB specimens and characterize the phenotype of B7H3 knock-down in NB cell line. Methods : Immunohistochemical (IHC) staining was carried out to assess the expression of B7H3 in clinical NB specimens. Survival association was analyzed using five Gene Expression Omnibus (GEO) datasets (GSE85047, GSE45480, GSE62564, GSE16476, GSE49710). Clonogenic survival and flow cytometry were performed after B7H3 knockdown to assess the cellular proliferation and cell survival in vitro. Impact of B7H3 silencing on NB growth was examined in vivo using the SH-SY5Y xenograft model. Results : On IHC staining, B7H3 was widely expressed in clinical NB specimens. Analysis of the transcriptional profiles of five GEO datasets clinically annotated NB specimens revealed that decreased B7H3 expression was associated with improved overall survival. B7H3 knockdown suppressed the proliferation of the SH-SY5Y NB model in vitro and in vivo. Cell cycle analysis revealed that B7H3 silencing induced G1/S arrest. This arrest was associated with the suppression of E2F1 expression and induction of Rb expression. Conclusion : Our results demonstrate that B7H3 expression correlate with clinical survival in NB patients. Preliminary studies suggest that B7H3 may mediate the G1/S transition.

The Convolution Sum $\sum_{al+bm=n}{\sigma}(l){\sigma}(m)$ for (a, b) = (1, 28),(4, 7),(1, 14),(2, 7),(1, 7)

  • Alaca, Ayse;Alaca, Saban;Ntienjem, Ebenezer
    • Kyungpook Mathematical Journal
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    • v.59 no.3
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    • pp.377-389
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    • 2019
  • We evaluate the convolution sum $W_{a,b}(n):=\sum_{al+bm=n}{\sigma}(l){\sigma}(m)$ for (a, b) = (1, 28),(4, 7),(2, 7) for all positive integers n. We use a modular form approach. We also re-evaluate the known sums $W_{1,14}(n)$ and $W_{1,7}(n)$ with our method. We then use these evaluations to determine the number of representations of n by the octonary quadratic form $x^2_1+x^2_2+x^2_3+x^2_4+7(x^2_5+x^2_6+x^2_7+x^2_8)$. Finally we express the modular forms ${\Delta}_{4,7}(z)$, ${\Delta}_{4,14,1}(z)$ and ${\Delta}_{4,14,2}(z)$ (given in [10, 14]) as linear combinations of eta quotients.

Biological Synthesis of 7-O-Methyl Apigenin from Naringenin Using Escherichia coli Expressing Two Genes

  • Jeon, Young-Min;Kim, Bong-Gyu;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.491-494
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    • 2009
  • Within the secondary metabolite class of flavonoids, which consist of more than 10,000 known structures, flavones define one of the largest subgroups. The diverse function of flavones in plants as well as their various roles in the interaction with other organisms offers many potential applications including in human nutrition and pharmacology. We used two genes, flavone synthase (PFNS-l) that converts naringenin into apigenin and flavone 7-O-methyltransferase (POMT-7) that converts apigenin into 7-O-methyl apigenin, to synthesize 7-O-methyl apigenenin from naringenin. The PFNS-l gene was subcloned into the E. coli expression vector pGEX and POMT-7 was subcloned into the pRSF vector. Since both constructs contain different replication origins and selection markers, they were cotransformed into E. coli. Using E. coli transformants harboring both PFNS-l and POMT-7, naringenin could be converted into 7-O-methyl apigenin, genkwanin.

Synthesis and in vitro Antitumor Activity of 2-Alkyl, 2-Aryl, and 2-Piperazinyl Benzimidazole-4, 7-dione Derivatives

  • Ahn, Chan-Mug;Tak, Jung-Ae;Choi, Sun-Ju
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.288-301
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    • 2000
  • A series of 2-alkyl, 2-aryl, and 2-piperazinyl benzimidazole-4,7-dione derivatives (7a-h) and 16m-o) were prepared, and their cytotoxicities were tested against three cancer cell lines (mouse lymphocytic leukemia cell line P388, and human gastric carcinoma cell lines SNU-1 and SNU-16). These compounds showed potent cytotoxicity against all of three cell lines tested, and especially SNU-16 was sensitive to them. 2-Aryl (7g,h) and 2-piperazinyl benzimidazole-4,7-dione derivative (I6 m) were more potent than mitomycin C against P388 and SNU-16. Among benzimidazole-4,7-dione derivatives with alkyl group at position 2, 7a had the most potent cytotoxicity against all of the cell lines tested.

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First detection and genetic characterization of porcine parvovirus 7 from Korean domestic pig farms

  • Ouh, In-Ohk;Park, Seyeon;Lee, Ju-Yeon;Song, Jae Young;Cho, In-Soo;Kim, Hye-Ryung;Park, Choi-Kyu
    • Journal of Veterinary Science
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    • v.19 no.6
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    • pp.855-857
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    • 2018
  • Porcine parvovirus 7 (PPV7) was first detected in Korean pig farms in 2017. The detection rate of PPV7 DNA was 24.0% (30/125) in aborted pig fetuses and 74.9% (262/350) in finishing pigs, suggesting that PPV7 has circulated among Korean domestic pig farms. Phylogenetic analysis based on capsid protein amino acid sequences demonstrated that the nine isolated Korean strains (PPV-KA1-3 and PPV-KF1-6) were closely related to the previously reported USA and Chinese PPV7 strains. In addition, the Korean strains exhibit genetic diversity with both insertion and deletion mutations. This study contributes to the understanding of the molecular epidemiology of PPV7 in Korea.

Microstructure and Electrical Properties of ZPCCYT Varistor Ceramics

  • Nahm, Choon-Woo;Lee, Sun-Kwon
    • Transactions on Electrical and Electronic Materials
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    • v.13 no.5
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    • pp.262-265
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    • 2012
  • The Microstructure and nonlinear electrical properties of the ZPCCYT (ZnO-$Pr_6O_{11}$-CoO-$Cr_2O_3$-$Y_2O_3$-$Tb_4O_7$) varistors were investigated for different amounts of $Tb_4O_7$. The addition of $Tb_4O_7$ has a significant effect on microstructure and electrical properties. Analysis of the microstructure indicated that the ceramics consisted of ZnO grain as a main phase and a few secondary phases as a mix of $Pr_6O_{11}$, $Y_2O_3$, and $Tb_4O_7$. As the amount of $Tb_4O_7$ increased, the sintered densities of pellets increased from $\rho$ = 5.70 to $5.78g/cm^3$ and the average grain size decreased from d = 4.8 to $3.6{\mu}m$. The increase in the amount of $Tb_4O_7$ increased from $E_B$ = 7,473 to 10,035 V/cm and from ${\alpha}$ = 39.7 to 52.2. In particular, it was found that the ceramics modified with 1.0 mol% in the amount of $Tb_4O_7$ are suited for the varistors for high voltage in the light of a high sintered density and a high voltage gradient.

Protocadherin-7 contributes to maintenance of bone homeostasis through regulation of osteoclast multinucleation

  • Kim, Hyunsoo;Takegahara, Noriko;Walsh, Matthew C.;Ueda, Jun;Fujihara, Yoshitaka;Ikawa, Masahito;Choi, Yongwon
    • BMB Reports
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    • v.53 no.9
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    • pp.472-477
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    • 2020
  • Osteoclasts are hematopoietic-derived cells that resorb bone. They are required to maintain proper bone homeostasis and skeletal strength. Although osteoclast differentiation depends on receptor activator of NF-κB ligand (RANKL) stimulation, additional molecules further contribute to osteoclast maturation. Here, we demonstrate that protocadherin-7 (Pcdh7) regulates formation of multinucleated osteoclasts and contributes to maintenance of bone homeostasis. We found that Pcdh7 expression is induced by RANKL stimulation, and that RNAi-mediated knockdown of Pcdh7 resulted in impaired formation of osteoclasts. We generated Pcdh7-deficient mice and found increased bone mass due to decreased bone resorption but without any defect in bone formation. Using an in vitro culture system, it was revealed that formation of multinucleated osteoclasts is impaired in Pcdh7-deficient cultures, while no apparent defects were observed in differentiation and function of Pcdh7-deficient osteoblasts. Taken together, these results reveal an osteoclast cell-intrinsic role for Pcdh7 in maintaining bone homeostasis.

Transient receptor potential melastatin type 7 channels are involved in zinc-induced apoptosis in gastric cancer

  • Kim, Byung-Joo
    • Animal cells and systems
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    • v.15 no.2
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    • pp.123-130
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    • 2011
  • Transient receptor potential melastatin 7 (TRPM7) channels are novel $Ca^{2+}$-permeable non-selective cation channels that are ubiquitously expressed. Activation of TRPM7 channels has been shown to be involved in the survival of gastric cancer cells. Here we show evidence suggesting that TRPM7 channels play an important role in $Zn^{2+}$- mediated cellular injury. Using a combination of electrophysiology, pharmacological analysis, small interfering RNA (siRNA) methods and cell death assays, we showed that activation of TRPM7 channels augmented $Zn^{2+}$-induced apoptosis of AGS cells, the most common human gastric adenocarcinoma cell line. The $Zn^{2+}$-mediated cytotoxicity was inhibited by the non-specific TRPM7 blockers $Gd^{3+}$ or 2 aminoethoxydiphenyl borate (2-APB) and TRPM7 specific siRNA. In addition, we showed that overexpression of TRPM7 channels in HEK293 cells increased $Zn^{2+}$- induced cell injury. Thus, TRPM7 channels may represent a novel target for physiological disorders where $Zn^{2+}$ toxicity plays an important role.