• Journal of Microbiology and Biotechnology
• /
• v.20 no.1
• /
• pp.45-53
• /
• 2010

We have developed an efficient and precise method for genome manipulations in Bacillus subtilis that allows rapid alteration of a gene sequence or multiple gene sequences without altering the chromosome in any other way. In our approach, the Escherichia coli toxin gene mazF, which was used as a counter-selectable marker, was placed under the control of a xylose-inducible expression system and associated with an antibiotic resistance gene to create a "mazF-cassette". A polymerase chain reaction (PCR)-generated fragment, consisting of two homology regions joined to the mazF-cassette, was integrated into the chromosome at the target locus by homologous recombination, using positive selection for antibiotic resistance. Then, the excision of the mazF-cassette from the chromosome by a single-crossover event between two short directly repeated (DR) sequences, included in the design of the PCR products, was achieved by counter-selection of mazF. We used this method efficiently and precisely to deliver a point mutation, to inactivate a specific gene, to delete a large genomic region, and to generate the in-frame deletion with minimal polar effects in the same background.

• Korean Journal of Cognitive Science
• /
• v.28 no.4
• /
• pp.329-347
• /
• 2017

Early research into category-based feature inference reported various phenomena in human thinking including typicality, diversity, similarity effects, etc. Later research discovered that participants' prior knowledge has an extensive influence on these sorts of reasoning. The current research tested the effects of causal knowledge on feature inference and conducted modeling on the results. Participants performed feature inference for categories consisted of four features where the features were connected either in common cause or common effect structure. The results showed typicality effects along with violations of causal Markov condition in common cause structure and causal discounting in common effect structure. To model the results, it was assumed that participants perform feature inference based on the difference between the probabilities of an exemplar with the target feature and an exemplar without the target feature (that is, $p(E_{F(X)}{\mid}Cat)-p(E_{F({\sim}X)}{\mid}Cat)$). Exemplar probabilities were computed based on causal model theory (Rehder, 2003) and applied to inference for target features. The results showed that the model predicts not only typicality effects but also violations of causal Markov condition and causal discounting observed in participants' data.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.11
• /
• pp.4509-4513
• /
• 2015

Osteosarcoma is the most common primary bone tumor in humans, especially in childhood. However, the genetic etiology for its pathogenesis remains elusive. It is known that microRNAs (miRNAs) are involved in the development of tumor progression. Here we show that microRNA-9 (miR-9) is a potential oncogene upregulated in osteosarcoma cells. Knockdown of miR-9 in osteosarcoma resulted in suppressed colony formation and cell proliferation. Further study identified GCIP, a Grap2 and cyclin D interacting protein, as a direct target of miR-9. In addition, GCIP overexpression activated retinoblastoma 1 (Rb) and suppressed E2F transcriptional target expression in osteosarcoma cells. Moreover, GCIP depletion reversed miR-9 knockdown induced colony formation and cell proliferation suppression. In sum, these results highlight the importance of miR-9 as an oncogene in regulating the proliferation of osteosarcoma by directly targeting GCIP and may provide new insights into the pathogenesis of osteosarcoma.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.2
• /
• pp.937-943
• /
• 2014

Polymorphisms in miRNA binding sites have been shown to affect miRNA binding to target genes, resulting in differential mRNA and protein expression and susceptibility to common diseases. Our purpose was to predict SNPs (single nucleotide polymorphisms) within miRNA binding sites of inflammatory genes in relation to gastric cancer. A complete list of SNPs in the 3'UTR regions of all inflammatory genes associated with gastric cancer was obtained from Pubmed. miRNA target prediction databases (MirSNP, Targetscan Human 6.2, PolymiRTS 3.0, miRNASNP 2.0, and Patrocles) were used to predict miRNA target sites. There were 99 SNPs with MAF>0.05 within the miRNA binding sites of 41 genes among 72 inflammation-related genes associated with gastric cancer. NF-${\kappa}B$ and JAK-STAT are the two most important signaling pathways. 47 SNPs of 25 genes with 95 miRNAs were predicted. CCL2 and IL1F5 were found to be the shared target genes of hsa-miRNA-624-3p. Bioinformatic methods could identify a set of SNPs within miRNA binding sites of inflammatory genes, and provide data and direction for subsequent functional verification research.

• The Plant Pathology Journal
• /
• v.35 no.5
• /
• pp.393-405
• /
• 2019

Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
• /
• 1999.11a
• /
• pp.228-231
• /
• 1999

The preparation and electrical properties of LIPON electrolyte were investigated in order to fabricate all solid state thin film battery. The LIPON thin film was deposited by r.f. sputtering of Li$_3$PO$_4$ target in O$_2$-N$_2$ mixtures. The LIPON deposited at N$_2$+10% O$_2$ ratio had a conductivity at 25 $^{\circ}C$ of 1.8${\times}$10$\^$-6/S/cm. The ion conductivity of the LIPON films decreased as the O$_2$ content of the process gas increased.

• Journal of Ginseng Research
• /
• v.47 no.1
• /
• pp.106-116
• /
• 2023

Background: Pirarubicin (THP) is an anthracycline antibiotic used to treat various malignancies in humans. The clinical usefulness of THP is unfortunately limited by its dose-related cardiotoxicity. Ginsenoside F1 (GF1) is a metabolite formed when the ginsenosides Re and Rg1 are hydrolyzed. However, the protective effects and underlying mechanisms of GF1 on THP-induced cardiotoxicity remain unclear. Methods: We investigated the anti-apoptotic and anti-oxidative stress effects of GF1 on an in vitro model, using H9c2 cells stimulated by THP, plus trigonelline or AKT inhibitor imidazoquinoxaline (IMQ), as well as an in vivo model using THP-induced cardiotoxicity in rats. Using an enzyme-linked immunosorbent test, the levels of malondialdehyde (MDA), brain natriuretic peptide (BNP), creatine kinase (CK-MB), cardiac troponin (c-TnT), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) were determined. Nuclear factor (erythroid-derived2)-like 2 (Nrf2) and the expression of Nrf2 target genes, including heme oxygenase-1 (HO-1), glutathione-S-transferase (Gst), glutamate-cysteine ligase modifier subunit (GCLM), and expression levels of AKT/Bcl-2 signaling pathway proteins were detected using Western blot analysis. Results: THP-induced myocardial histopathological damage, electrocardiogram (ECG) abnormalities, and cardiac dysfunction were reduced in vivo by GF1. GF1 also decreased MDA, BNP, CK-MB, c-TnT, and LDH levels in the serum, while raising SOD and GSH levels. GF1 boosted Nrf2 nuclear translocation and Nrf2 target gene expression, including HO-1, Gst, and GCLM. Furthermore, GF1 regulated apoptosis by activating AKT/Bcl-2 signaling pathways. Employing Nrf2 inhibitor trigonelline and AKT inhibitor IMQ revealed that GF1 lacked antioxidant and anti-apoptotic effects. Conclusion: In conclusion, GF1 was found to alleviate THP-induced cardiotoxicity via modulating Nrf2 and AKT/Bcl-2 signaling pathways, ultimately alleviating myocardial oxidative stress and apoptosis.

• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.4
• /
• pp.571-579
• /
• 2016

In this work, we prepared a panel of monoclonal anti-idiotypic antibodies to ovine prolactin (oPRL) by the hybridoma technique. Among these antibodies, one anti-idotypic antibody (designated B7) was chosen for further characterization by a series of experiments. We first demonstrated that B7 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assays. Subsequently, the results of a competitive receptor-binding assay confirmed that B7 could specifically bind to the prolactin receptor (PRLR) expressed on target cells. Finally, we examined its biological activities in CHO-PRLR and Nb2 cells and observed that B7 could activate Janus kinase 2-signal transducer and activator of transcription signalling in CHO-PRLR and Nb2 cells and induce BaF3 proliferation. The present study suggests that i) B7 can serve as a PRLR agonist or PRL mimic and has potential applications in regulating mammary gland development, milk production and maintenance of lactation in domestic animals and ii) B7 may be a biological reagent that can be used to explore the mechanism of PRLR-mediated intracellular signalling.

• Economic and Environmental Geology
• /
• v.28 no.3
• /
• pp.259-269
• /
• 1995

Column leaching tests were conducted using fresh and weathered pulverised fuel ash of some 17 and 40 years old from two major British power plants, with deionised water and simulated synthetic industrial leachate. The former was to see the leaching behaviour of weathered ash and the latter was to see if the formation of secondary products from water and PFA interaction and ameliorating effect in removing metals from industrial leachates. Fresh PFA liberates elevated concentrations of surface-enriched inorganics, including Ca, Na, K, B, $Cr_{total}$, Li Mo, Se and $SO^{2-}_4$. This might indicate their association with the surface of PFA particles. In the column leaching tests using weathered ash and deionised water, elements are not readily leached but are released more slowly, showing relatively constant concentrations. For the case of weathered ash, some readily soluble surface-enriched elements appears to have been liberated in their early stage of leaching and the liberation of glass associated elements are thought to be more important function in controlling the element concentration. The result from column leaching tests exceed for a number of elements when compared with various Water Standards and suggests the leachate from PFA disposal mound needs dilution to achieve target concentrations. PF A shows element retention effect for many elements, including B, Fe, Zn, Hg, Ni, Li and Mo, in the order of fresh Drax ash > weathered Drax ash > Weathered Meaford ash in retaining capacity. Geochemical modelling using a computer program WATEQ4F reveals some solubility controlling secondary solid products. These include $CaSO_4{\cdot}2H_2O$ for Ca, $Al(OH)_3$ for Al and $Fe(OH)_3$ for Fe.

• Nuclear Engineering and Technology
• /
• v.53 no.8
• /
• pp.2682-2695
• /
• 2021

This paper aims to evaluate the structural dynamic responses and damage/failure of the nuclear fuel reprocessing plant under the free drop impact of spent fuel cask (SFC) and fuel assembly (FA) during the on-site transportation. At the present Part I of this paper, the large-scale SFC model free drop test and the corresponding numerical simulations are performed. Firstly, a composite target which is composed of the protective structure, i.e., a thin RC plate (representing the inverted U-shaped slab in the loading shaft) and/or an autoclaved aerated concrete (AAC) blocks sacrificial layer, as well as a thick RC plate (representing the bottom slab in the loading shaft) is designed and fabricated. Then, based on the large dropping tower, the free drop test of large-scale SFC model with the mass of 3 t is carried out from the height of 7 m-11 m. It indicates that the bottom slab in the loading shaft could not resist the free drop impact of SFC. The composite protective structure can effectively reduce the damage and vibrations of the bottom slab, and the inverted U-shaped slab could relieve the damage of the AAC blocks layer dramatically. Furthermore, based on the finite element (FE) program LS-DYNA, the corresponding refined numerical simulations are performed. By comparing the experimental and numerical damage and vibration accelerations of the composite structures, the present adopted numerical algorithms, constitutive models and parameters are validated, which will be applied in the further assessment of drop impact effects of full-scale SFC and FA on prototype nuclear fuel reprocessing plant in the next Part II of this paper.