• Title/Summary/Keyword: CO I gene

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Characterization of a conjugated polysuccinimide-carboplatin compound

  • Sun Young Lee;Chang Hoon Chae;Miklos Zrinyi;Xiangguo Che;Je Yong Choi;Dong-Hyu Cho
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.1
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    • pp.31-38
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    • 2023
  • Carboplatin, an advanced anticancer drug with excellent efficacy against ovarian cancer, was developed to alleviate the side effects that often occur with cisplatin and other platinum-based compounds. Our study reports the in vitro characteristics, viability, and activity of cells expressing the inducible nitric oxide synthase (iNOS) gene after carboplatin was conjugated with polysuccinimide (PSI) and administered in combination with other widely used anticancer drugs. PSI, which has promising properties as a drug delivery material, could provide a platform for prolonging carboplatin release, regulating its dosage, and improving its side effects. The iNOS gene has been shown to play an important role in both cancer cell survival and inhibition. Herein, we synthesized a PSI-carboplatin conjugate to create a modified anticancer agent and confirmed its successful conjugation. To ensure its solubility in water, we further modified the structure of the PSI-carboplatin conjugate with 2-aminoethanol groups. To validate its biological characteristics, the ovarian cancer cell line SKOV-3 and normal ovarian Chinese hamster ovary cells were treated with the PSI-carboplatin conjugate alone and in combination with paclitaxel and topotecan, both of which are used in conventional chemotherapy. Notably, PSI-carboplatin conjugation can be used to predict changes in the genes involved in cancer growth and inhibition. In conclusion, combination treatment with the newly synthesized polymer-carboplatin conjugate and paclitaxel displayed anticancer activity against ovarian cancer cells but was not toxic to normal ovarian cancer cells, resulting in the development of an effective candidate anticancer drug without severe side effects.

Adsorption Characteristics of Strong Basic Anion Exchanger to Cellulose Reactive Dye (강 염기성 음이온 교환수지의 셀룰로우스 섬유용 반응성 염료에 대한 흡착 특성)

  • Lim, Gyeong-Eun;Chung, Paul-gene;Kwon, Ji-Young
    • Journal of Korean Society on Water Environment
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    • v.23 no.1
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    • pp.27-32
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    • 2007
  • This study focused on estimating the feasibility of a strong basic anion exchanger (PA312OH) as a sorbent for the removal of residual reactive dye and saving chemicals and water. Cellulose reactive dye C.I.RB49 was tested because reactive dye is the largest single group of dyes and that dye needs larger amount of inorganic salts as dyeing agent but nearly 50% of reactive dyes may be lost to the effluent. The adsorption characteristics of PA312OH for C.I.RB49 were as follows. Ion-selectivity among the dye and inorganic salts was Dye > ${SO_4}^{2-}$ > ${CO_3}^{2-}$ > $Cl^-$. C.I.RB49 was exchanged more than 3 times ${SO_4}^{2-}$ and ${CO_3}^{2-}$ and $Cl^-$ was not exchanged absolutely. The exchanging velocity was increased exponentially with increasing temperature. This result is positive effect on treating the high temperature dyeing process wastewater. The exchanged dye percents to initial were 96.8% and 99% at flow rate 20.5 mL/min. and 3.7 mL/min.. The exchanging capacity of PA312OH for C.I.RB49 was 215.2 mg/g at conc.=369.2 mg/L, Temp.=$25^{\circ}C$. 74% inorganic salts were recrystallized from real dark reactive color dyeing wastewater treated with PA312OH.

Effective Application of Multiplex RT-PCR for Characterization of Human Embryonic Stem Cells/ Induced Pluripotent Stem Cells (다중 역전사 중합효소 연쇄 반응(Multiplex RT-PCR)을 이용한 인간배아 줄기세포 및 유도만능 줄기세포의 효과적인 분화 양상 조사)

  • Kim, Jung-Mo;Cho, Youn-Jeong;Son, On-Ju;Hong, Ki-Sung;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.1-8
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    • 2011
  • Techniques to evaluate gene expression profiling, such as sufficiently sensitive cDNA microarrays or real-time quantitative PCR, are efficient methods for monitoring human pluripotent stem cell (hESC/iPSC) cultures. However, most of these high-throughput tests have a limited use due to high cost, extended turn-around time, and the involvement of highly specialized technical expertise. Hence, there is an urgency of rapid, cost-effective, robust, yet sensitive method development for routine screening of hESCs/hiPSCs. A critical requirement in hESC/hiPSC cultures is to maintain a uniform undifferentiated state and to determine their differentiation capacity by showing the expression of gene markers representing all three germ layers, including ectoderm, mesoderm, and endoderm. To quantify the modulation of gene expression in hESCs/hiPSC during their propagation, expansion, and differentiation via embryoid body (EB) formation, we developed a simple, rapid, inexpensive, and definitive multimarker, semiquantitative multiplex RT-PCR platform technology. Among the 9 gene primers tested, 5 were pluripotent markers comprising set 1, and 3 lineage-specific markers were combined as set 2, respectively. We found that these 2 sets were not only effective in determining the relative differentiation in hESCs/hiPSCs, but were easily reproducible. In this study, we used the hES/hiPS cell lines to standardize the technique. This multiplex RT-PCR assay is flexible and, by selecting appropriate reporter genes, can be designed for characterization of different hESC/hiPSC lines during routine maintenance and directed differentiation.

Molecular Cloning and Nucleotide Sequence of the G protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus (한국에서 분리된 IHNV-PRT의 G protein의 유전자 클로닝과 염기서열 분석)

  • 김영조;허강준;박정우;박정문
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.226-230
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    • 1999
  • To characterize the Korean isolate of infeciious hematopoietic necrosis virus (IHNV-PRT), a partial DNA fragment G gene of the MNV-PRT was amplified by RT-PCR. cloned inlo pGEM-T easy vector and analyzed for nucleotlde sequences. The size of the PCR pmduct was about 442 bp. The nucleotlde sequence homologies ofthe G gene of IHNV-PRT were 95%, 94%, 94% 94%, 93%, 53%. respectively. with those of foreign isolates of IHNV, IHNV-RB-76. IHNV-LR-73, MNV-K, IHNV-WRAC, Im-SRCV, IHNV-Col-85. However, it showed 81% homology with that of other fish rhabdovirus, hisame rhabdovirus (HRV). Frou~ the rcsults of deduced amino acid sequence homology analysis. G protein of IHNV-PRT showed 96% hornologies with those of foreign isolates of IHNV but 89% homology with that of HRV These results indicaled that, even though G gene of IHIW-PRT showed low homology with that of HRY it was highly conserved among different strains of THNV.

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HPAI-resistant Ri chickens exhibit elevated antiviral immune-related gene expression

  • Thi Hao Vu;Jubi Heo;Yeojin Hong;Suyeon Kang;Ha Thi Thanh Tran;Hoang Vu Dang;Anh Duc Truong;Yeong Ho Hong
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.13.1-13.11
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    • 2023
  • Background: Highly pathogenic avian influenza viruses (HPAIVs) is an extremely contagious and high mortality rates in chickens resulting in substantial economic impact on the poultry sector. Therefore, it is necessary to elucidate the pathogenic mechanism of HPAIV for infection control. Objective: Gene set enrichment analysis (GSEA) can effectively avoid the limitations of subjective screening for differential gene expression. Therefore, we performed GSEA to compare HPAI-infected resistant and susceptible Ri chicken lines. Methods: The Ri chickens Mx(A)/BF2(B21) were chosen as resistant, and the chickens Mx(G)/BF2(B13) were selected as susceptible by genotyping the Mx and BF2 genes. The tracheal tissues of HPAIV H5N1 infected chickens were collected for RNA sequencing followed by GSEA analysis to define gene subsets to elucidate the sequencing results. Results: We identified four differentially expressed pathways, which were immune-related pathways with a total of 78 genes. The expression levels of cytokines (IL-1β, IL-6, IL-12), chemokines (CCL4 and CCL5), type interferons and their receptors (IFN-β, IFNAR1, IFNAR2, and IFNGR1), Jak-STAT signaling pathway genes (STAT1, STAT2, and JAK1), MHC class I and II and their co-stimulatory molecules (CD80, CD86, CD40, DMB2, BLB2, and B2M), and interferon stimulated genes (EIF2AK2 and EIF2AK1) in resistant chickens were higher than those in susceptible chickens. Conclusions: Resistant Ri chickens exhibit a stronger antiviral response to HPAIV H5N1 compared with susceptible chickens. Our findings provide insights into the immune responses of genetically disparate chickens against HPAIV.

Enhanced Production of Astaxanthin by Archaea Chaperonin in Escherichia coli (대장균에서 고세균 샤페론을 이용한 아스타잔틴 생산능 향상을 위한 연구)

  • Seo, Yong Bae;Lee, Jong Kyu;Jeong, Tae Hyug;Nam, Soo-Wan;Kim, Gun-Do
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1339-1346
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    • 2015
  • The aim of this study is to increase production of carotenoids in recombinant Escherichia coli by Archaea chaperonin. The carotenoids are a widely distributed class of structurally and functionally diverse yellow, orange, and red natural pigments. These pigments are synthesized in bacteria, algae, fungi, and plants, and have been widely used as a feed supplement from poultry rearing to aquaculture. Carotenoids also exhibit diverse biological properties, such as strong antioxidant and antitumor activities, and enhancement of immune responses. In the microbial world, carotenoids are present in both anoxygenic and oxygenic photosynthetic bacteria and algae and in many fungi. We have previously reported cloning and functional analysis of the carotenoid biosynthesis genes from Paracoccus haeundaensis. The carotenogenic gene cluster involved in astaxanthin production contained seven carotenogenic genes (crtE, crtB, crtI, crtY, crtZ, crtW and crtX genes) and recombinant Escherichia coli harboring seven carotenogenic genes from Paracoccus haeundaensis produced 400 μg/g dry cell weight (DCW) of astaxanthin. In order to increase production of astaxanthin, we have co-expressed chaperone genes (ApCpnA and ApCpnB) in recombinant Escherichia coli harboring the astaxanthin biosynthesis genes. This engineered Escherichia coli strain containing both chaperone gene and astaxanthin biosynthesis gene cluster produced 890 μg/g DCW of astaxanthin, resulting 2-fold increased production of astaxanthin.

Authentication of Salted-dried Fish Species Using Polymerase Chain Reaction-Single Strand Conformational Polymorphism and Restriction Analysis of Mitochondrial DNA

  • Kim, Joo-Shin;Chu, Kin Kan Astley;Kwan, Hoi Shan;Chung, Hau Yin
    • Fisheries and Aquatic Sciences
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    • v.11 no.3
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    • pp.133-139
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    • 2008
  • Molecular techniques, including restriction fragment length polymorphism(RFLP) and polymerase chain reaction-single strand conformational polymorph isms(PCR-SSCP), were developed to identify salted, dried threadfin(Eleutheronema tetradactylum) and white herring(Ilisha elongata) fish. Using PCR with universal primers, conserved 367-bp fragments of the cytochrome b gene were amplified from fresh fish samples and sequenced. The sequences were then searched for specific restriction sites. The digestion of the PCR products with the endonucleases AvaI, FokI, MboII, and MspI generated RFLP, which was used to identify the commercial products. Similarly, the amplified PCR-SSCP products were developed and the products tested. Overall, similar patterns were found in the majority of the fresh and processed products. Based on the results, both RFLP and PCR-SSCP were useful in determining and validating the authenticity of the fish species used to prepare the commercial salted, dried products. A similar approach can be applied to other species.

Gene Duplications Revealed during the Process of SNP Discovery in Soybean[Glycine max(L.) Merr.]

  • Cai, Chun Mei;Van, Kyu-Jung;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.237-242
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    • 2007
  • Genome duplication(i.e. polyploidy) is a common phenomenon in the evolution of plants. The objective of this study was to achieve a comprehensive understanding of genome duplication for SNP discovery by Thymine/Adenine(TA) cloning for confirmation. Primer pairs were designed from 793 EST contigs expressed in the roots of a supernodulating soybean mutant and screened between 'Pureunkong' and 'Jinpumkong 2' by direct sequencing. Almost 27% of the primer sets were failed to obtain sequence data due to multiple bands on agarose gel or poor quality sequence data from a single band. TA cloning was able to identify duplicate genes and the paralogous sequences were coincident with the nonspecific peaks in direct sequencing. Our study confirmed that heterogeneous products by the co-amplification of a gene family member were the main cause of obtaining multiple bands or poor quality sequence data in direct sequencing. Counts of amplified bands on agarose gel and peaks of sequencing trace suggested that almost 27% of nonrepetitive soybean sequences were present in as many as four copies with an average of 2.33 duplications per segment. Copy numbers would be underestimated because of the presence of long intron between primer binding sites or mutation on priming site. Also, the copy numbers were not accurately estimated due to deletion or tandem duplication in the entire soybean genome.

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Study on the Control of Biofilm Formation Inhibition on Pantoea agglomerans by Anti-bacterial Effect of Indole (인돌의 항균 효과에 의한 Pantoea agglomerans의 바이오필름 생성 억제 조절에 관한 연구)

  • Jin, Seul;Yang, Woong-Suk;Hwang, Cher-Won;Lee, Jae-Yong
    • Journal of Environmental Science International
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    • v.30 no.5
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    • pp.369-378
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    • 2021
  • In this study, we investigated the effects of indole on biofilm formation inhibition in Pantoea agglomerans (P. agglomerans). In the biofilm growth assay, indole inhibited biofilm formation across all the growth time. Depending on biofilm growth stage, indole exhibited biofilm inhibition and anti-bacterial effects on planktonic cells. Through the analysis of the proportion rate between biofilm and Colony Forming Units (CFU) and inhibition rate of indole, we confirmed that depending on the biofilm stage of P. agglomerans, indole treatment timing was more important than the treatment duration. By comparing gene expression rates through rt-qPCR P.agglomerans affected by indole was found to significantly change quorum sensing (pagI/R) and indole transportation (bssS) gene expressions. Throughout all, indole exhibited both antimicrobial and anti-biofilm effects on P. agglomerans. In addition, we confirmed the anti-biofilm effects of indole on mature biofilm. In conclusion, indole as a signal molecule, can exhibit anti-biofilm effects through bacterial quorum sensing inhibition and indole affects. Therefore, indole can regulate biofilm bacteria especially gram-negative opportunistic pathogens.

Occurrence and Molecular Identification of Microcotyle sebastis Isolated from Fish Farms of the Korean Rockfish, Sebastes schlegelii

  • Song, Jun-Young;Kim, Keun-Yong;Choi, Seo-Woo
    • Parasites, Hosts and Diseases
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    • v.59 no.1
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    • pp.89-95
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    • 2021
  • Microcotyle sebastis is a gill monogenean ectoparasite that causes serious problems in the mariculture of the Korean rockfish, Sebastes schlegelii. In this study, we isolated the parasite from fish farms along the coasts of Tongyeong, South Korea in 2016, and characterized its infection, morphology and molecular phylogeny. The prevalence of M. sebastis infection during the study period ranged from 46.7% to 96.7%, and the mean intensity was 2.3 to 31.4 ind./fish, indicating that the fish was constantly exposed to parasitic infections throughout the year. Morphological observations under light and scanning electron microscopes of the M. sebastis isolates in this study showed the typical characteristics of the anterior prohaptor and posterior opisthaptor of monogenean parasites. In phylogenetic trees reconstructed using the nuclear 28S ribosomal RNA gene and the mitochondrial cytochrome c oxidase I gene (cox1), they consistently clustered together with their congeneric species, and showed the closest phylogenetic relationships to M. caudata and M. kasago in the cox1 tree.