• Title/Summary/Keyword: over-expression

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Molecular Mechanism of ABC Transporter Mdr49A Associated with a Positive Cross-Resistance in Transgenic Drosophila (형질전환 초파리를 이용한 Mdr49A 유전자의 살충제 교차저항성 기능 구명)

  • Seong, Keon Mook;Pittendrigh, Barry R.
    • Korean journal of applied entomology
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    • v.59 no.4
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    • pp.341-348
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    • 2020
  • The ATP-binding cassette (ABC) transporter superfamily represents the largest transmembrane protein that transports a variety of substrates across extra- and intra-cellular membranes. In insects, the ABC transporter proteins play crucial roles in insecticide resistance. To date, no studies have investigated the involvement of ABC transporter gene for cross-resistance to insecticide chemistries. Here, we studied such possible mechanisms against six conventional insecticides using transgenic Drosophila melanogaster strains carrying Mdr49 transcript variant A. For the 91-R and 91-C strains of Drosophila melanogaster, although they have a common origin, 91-R has been intensely selected with DDT for over 60 years, while 91-C has received no insecticide selection. Our transgenic analyses showed that overexpression of 91-R-MDR49 transcript variant A along with three amino acid variations can yield a relatively low degree of cross-resistance to carbofuran (2.0~6.7-fold) and permethrin (2.5~10.5-fold) but did not show cross-resistance to abamectin, imidacloprid, methoxychlor, and prothiofos as compared to the Gal4-driver control strain without transgene expression. These results indicate that the overexpression of Mdr49A in itself leads to a cross-resistance and three amino acid changes have additional effects on positive cross-resistance to carbofuran and permethrin.

Extracellular RNAs and Extracellular Vesicles: Inception, Current Explorations, and Future Applications

  • Perumal, Ayyappasamy Sudalaiyadum;Chelliah, Ramachandran;Datta, Saptashwa;Krishna, Jayachandran;Samuel, Melvin S.;Ethiraj, Selvarajan;Park, Chae Rin
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.535-543
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    • 2020
  • In addition to the ubiquitous roles of cellular RNA in genetic regulations, gene expression and phenotypic variations in response to environmental cues and chemotactic signals, the regulatory roles of a new type of RNA called extracellular RNAs (exRNAs) are an up-and-coming area of research interest. exRNA is transported outside the cell through membrane blebs known as membrane vesicles or extracellular vesicles (EVs). EV formation is predominant and conserved among all microbial forms, including prokaryotes, eukaryotes, and archaea. This review will focus on the three major topics concerning bacterially derived exRNAs, i.e., 1) the discovery of exRNA and influence of extraneous RNA over bacterial gene regulations, 2) the known secretion mechanism for the release of exRNA, and 3) the possible applications that can be devised with these exRNA secreted by different gram-negative and gram-positive bacteria. Further, this review will also provide an opinion on exRNA- and EV-derived applications such as the species-specific exRNA markers for diagnostics and the possible roles of exRNA in probiotics and the epigenetic regulations of the gut microbiome.

Evaluation of shrimp protein hydrolysate and krill meal supplementation in low fish meal diet for red seabream (Pagrus major)

  • Gunathilaka, Buddhi E.;Khosravi, Sanaz;Shin, Jaebeom;Shin, Jaehyeong;Herault, Mikael;Fournier, Vincent;Lee, Kyeong-Jun
    • Fisheries and Aquatic Sciences
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    • v.24 no.3
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    • pp.109-120
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    • 2021
  • Protein hydrolysates and krill meal (KM) are used as protein sources in aquafeeds. The study was conducted to examine the supplemental effects of shrimp protein hydrolysates (SH) or KM in a high-plant-protein diet for red seabream (Pagrus major). A fish meal (FM)-based diet (40%) was considered as the high-FM diet (HFM) and a diet containing 25% FM and soy protein concentrate, in the expense of FM protein from HFM diet, was considered as the low fish meal (LFM) diet. Two other experimental diets (SH and KM) were prepared by including SH and KM into LFM diet at 5% inclusion levels in exchange of 5% FM from the LFM diet. A feeding trial was conducted for fifteen weeks using triplicate group of fish (Initial mean body weight, 8.47 ± 0.05 g) for a diet. Growth performance and feed efficiency of fish were significantly enhanced by HFM, KM and SH supplemented diets over those of fish fed LFM diet. Interestingly, these parameters of fish fed SH diet showed better performance than KM and HFM groups. Liver IGF-I expression of fish fed SH diet was comparable to HFM group and higher than KM and LFM diets. Protein digestibility of SH diet was significantly higher than KM, HFM, and LFM diets. Dry matter digestibility of SH diet was comparable to HFM diet and significantly higher than KM and LFM diets. Nitro blue tetrazolium and superoxide dismutase activities of HFM, SH and KM groups were significantly elevated than the LFM group and SH diet increased catalase and glutathione peroxidase activities of fish compared to KM and LFM groups. Hemoglobin level and hematocrit of fish fed SH and KM diets were significantly higher than LFM group. A diet containing 20% FM with KM is comparable to a HFM diet which contains 40% FM for red seabream. SH can be used to replace FM from red seabream diet down to 20% and fish performance can be improved better than a diet containing 40% FM. Overall, it seems that SH is more effective ingredient in red seabream diet compared to KM.

Conservative Genes among 1,309 Species of Prokaryotes (원핵생물 1,309종의 보존적 유전자)

  • Lee, Dong-Geun
    • Journal of Life Science
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    • v.32 no.6
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    • pp.463-467
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    • 2022
  • As a result of applying the COG (Cluster of Orthologous Groups of Protein) algorithm to 1,309 species to confirm the conserved genes of prokaryotes, ribosomal protein S11 (COG0100) was identified. The numbers of conservative genes were 2, 5, 5, and 6 in 1,308, 1,307, 1,306, and 1,305 species, respectively. Twenty-nine genes were conserved in over 1,302 species, and they encoded 23 ribosomal proteins, 3 tRNA synthetases, 2 translation factors, and 1 RNA polymerase subunit. Most of them were related to protein production, suggesting the importance of protein expression in prokaryotes. The highest conservative COG was COG0048 (ribosomal protein S12) among the 29 COGs. The 29 conserved genes usually have one protein for each prokaryote. COG0090 (ribosomal protein L2) had not only the lowest conservation value but also the largest standard deviation of phylogenetic distance value. As COG0090 is not only a member of the ribosome, but also a regulator of replication and transcription, it could be inferred that prokaryotes have large variations in COG0090 to survive in various environments. This study could provide data necessary for basic science, tumor control, and development of antibacterial agents.

Severe choline deficiency induces alternative splicing aberrance in optimized duck primary hepatocyte cultures

  • Zhao, Lulu;Cai, Hongying;Wu, Yongbao;Tian, Changfu;Wen, Zhiguo;Yang, Peilong
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1787-1799
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    • 2022
  • Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

Effect of Gami-Chunggisan on Antioxidant and Pro-Inflammatory Cytokine (가미청기산(加味淸肌散)이 항산화와 염증성 사이토카인에 미치는 영향)

  • Lee, Youn-Jeong;Sim, Boo-Yong;Lee, Hae-Jin;Bak, Ji-Won;Kim, Dong-Hee
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.69-76
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    • 2014
  • Objectives : Gami-Chunggisan extract (GCE) is one of the oriental traditional medicine. We investigated the antioxidant effect and reduction of pro-inflammatory cytokine as a functional ingredient for cosmetic products from the GCE. Methods : GCE was prepared by extracting with 80% ethanol. We analyzed total polyphenol and antioxidant activities. To evaluate antioxidant activity, we measured 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay. Also we measured the production of reactive oxygen species (ROS) and nitric oxide (NO) on Raw264.7 cells. We researched reduction of anti-inflammatory cytokines from concentration of GCE on Raw264.7 cells. Results : Total polyphenol quantity of GCE was included 46.6 mg/g. The GCE showed ABTS free radical scavenging ability with more than 89% at $1000{\mu}g/m{\ell}$. In addition the DPPH free radical scavenging ability from the GCE was activated over 93% at $1000{\mu}g/m{\ell}$. Production of the ROS was decreased by approximately 26%, upon the GCE treatment at concentration of $100{\mu}g/m{\ell}$. The GCE at $100{\mu}g/m{\ell}$ concentration showed inhibitory effect on NO production by 38%. Production of IL-$1{\beta}$ and IL-6 were decreased by approximately 56% and 36%, respectively upon GCE treatment at $100{\mu}g/m{\ell}$. Also, production of TNF-${\alpha}$ was decreased by approximately 79% at $100{\mu}g/m{\ell}$. Moreover, the GCE showed inhibitory effects on the expression of the IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ genes in LPS-induced Raw 264.7 cells. Conclusions : From the results above, we conclude that the GCE indicated significant antioxidant effects and induced reduction of pro-inflammatory cytokine.

Effect of Cheongungdajosan-gamibang on oxidative damage and inflammation (천궁다조산가미방(川芎茶調散加味方) 주정추출물이 산화적 손상 및 염증에 미치는 영향)

  • Bak, Ji Won;Sim, Boo-Yong;Lee, Hae Jin;Kim, Dong Hee
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.53-59
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    • 2014
  • Objectives : The purpose of this study was verification of the anti-inflammation and anti-oxidant effect of Cheongungdajosan-gamibang extract (CG) in mouse macrophage, RAW 264.7 cells. Methods : We have basically using LPS-stimulated RAW 264.7 cells. The cell toxicity was determined by MTT assay. To evaluate the anti-inflammatory effect of Cheongungdajosan-gamibang, amount of nitric oxide(NO) was measured using the NO detection kit and the IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Also, free radical scavenging assay has tested for DPPH and ABTS radical activity as well as the contents of total polyphenol. Results : In this study, 96.6% or higher cell viability was observed in all tested groups from 1, 10, $100{\mu}/m{\ell}$ in RAW 264.7 cells. The RAW 264.7 cells were induced by lipopolysaccharide (LPS) and CG 1, 10, $100{\mu}/m{\ell}$. The CG decreased nitric oxide (NO) production activity dose dependently, especially at $100{\mu}/m{\ell}$ of 55%. The production of IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ were decreased by 51%, 78% and 35% in CG treated $100{\mu}g/m{\ell}$. CG showed dose-dependent suppression activity of reactive oxygen species (ROS) production, especially at $100{\mu}g/m{\ell}$ of 37%. DPPH radical scavenging activity and ABTS cation decolorization were activated over 86% and 88% in CG at $1,000{\mu}g/m{\ell}$ concentration. Conclusions : According to the results, we thought that CG showed anti-inflammatory and antioxidant activities on the RAW 264.7 cells in mouse macrophage. Therefore, this research is expected to provide the fundamental data about the natural material analysis of relating to the anti-inflammation and antioxidant.

A Study on the Verification of Physiological Activities of Ternstroemia kwangtungensis Merr. Extract (베트남 후피향 추출물의 생리활성 검증에 대한 연구)

  • Jung, Su-Hyun;Yeom, Hyeon-Ji;Oh, Min-Jeong;Lee, Jin-Young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.1
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    • pp.39-46
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    • 2022
  • In this study, we tried to find out the value of using Vietnamese Ternstroemia kwangtungensis Merr. (TK) extracts as a functional material. As a result of measuring the level of Nrf2 expression of TK, 50% ethanol extract TK leaf showed the highest activity at a concentration of 200 ㎍/mL. In order to confirm the stability of the extract over time at room temperature, UPLC was performed, and stability was maintained because the patterns of the UV spectrum pattern were the same. In order to confirm the antioxidant efficacy of TK, the activity increased as the concentration increased as a result of measuring electron donating and ABTS+ radical scavenging ability. At 1,000 ㎍/mL, they showed effects of 94.6% and 90.8%, respectively. As a result of measuring the tyrosinase inhibitory activity showed an inhibitory ability of 24.4% at a concentration of 1,000 ㎍/mL. In addition, as a result of examining the elastase inhibitory activity of the extract was increased in a concentration-dependent manner, and it was confirmed that the inhibitory activity was 42.7% at 1,000 ㎍/mL. From these results, it was confirmed that the TK extract was valuable as a natural material for cosmetics.

Comparison and evaluation of 89Zr-labeled trastuzumab and Thio-trastuzumab : a potential immuno-PET probe for HER2-positive carcinomas

  • Un Chol Shin;Seoku Bae;Suk-man Kim;Min-Woo Lee;Han Sang Jin;Hyun Park;Kyo Chul Lee;Jung Young Kim;Ji Woong Lee
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.7 no.2
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    • pp.105-111
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    • 2021
  • 89Zr is a positron-emitting radioisotope, which has known as well-suited radioisotope for use in a monoclonal antibody-based imaging agent for immuno-PET. The purpose of this study was to quantitatively evaluate the diagnostic ability of general trastuzumab and thio-trastuzumab as HER2 positive receptors based on Df hexadentate iron chelator. Desferrioxamine-p-SCN (Df-Bz-NCS) and desferroixamine-maleimide (Df-Mal) were purchased from Macrocyclics (Dallas, TX, USA). The trastuzumab was purchased from Roche (Schweiz), and thio-trastuzumab was obtained from professor Hyo-Jeong Hong group (Kangwon National University). The radioisotope 89Zr was produced by domestic purification system and KIRAMS using medical cyclotron (50 MeV, Scantronix). The conjugates of Df-trastuzumab and Df-thio-trastuzumab were prepared with Df-Bz-NCS and Df-Mal under basic aqueous solution (pH 8-9) at room temperature, respectively. The conjugates purified by PD-10 column were mixed with dried 89Zr chloride. 89Zr-labeled conjugates were purified and concentrated by Amicon ultra centrifugal filter. The preparation step and time of 89Zr-labeled conjugates was shorted as 4 steps within 2 hours. 89Zr-labeled conjugates showed the highly radiochemical purity of over 98%, and were very stable until 7 days by the analysis of radio-ITLC method. Each radio-labeled conjugates were also exhibited the highly stability in both PBS buffer and mouse serum. Immuno-PET imaging of 89Zr-labeled conjugates in mice bearing gastric cancer xenograft tumors with HER2 expression showed high tumor uptake in the NCI-N87 HER2-expressing. However, 89Zr-Df-Mal-thio-trastuzumab showed a relatively lower tumor-to-background ratio than 89Zr-Df-Bz-trastuzumab, as well as whole-body distribution. In the results, 89Zr-Df-Bz-trastuzumab was evaluated to have a relatively higher HER2 diagnostic ability than 89Zr-Df-Mal-thio-trastuzumab.

Cardioprotective Potential of Gracilaria Verrucosa Extract in Myocardial Infarction-Induced Heart Failure Model (심근 경색 유발 심부전 모델에서 강리 추출물의 심장 보호 가능성)

  • Youn Jae Jang;Hye Yoom Kim;Jung Joo Yoon;Byung Hyuk Han;Je Kuk Yu;Nam Geun Cho;Ho Sub Lee;Dae Gill Kang
    • Herbal Formula Science
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    • v.31 no.3
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    • pp.157-169
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    • 2023
  • Gracilaria Verrucosa (GV), a seaweed used in traditional Korean medicine, was studied for its effects on MI-induced heart failure in rats. MI is caused by a blocked coronary artery, leading to severe cardiac dysfunction. The study used a rat model to assess cardiac changes over time and evaluate the impact of GV on heart failure. Ischemia was induced through LAD ligation surgery, and the extent of ischemic area was measured as a prognostic factor. GV extract administration significantly improved cardiac morphology and reduced cardiac weight compared to the MI group. GV treatment also improved cardiac function, as evidenced by positive effects on chamber dilation during MI-induced heart failure. Parameters such as ejection fraction (EF) and fractional shortening (FS) were measured. The MI group showed decreased EF and FS compared to the sham group, while these parameters improved in the GV group. GV treatment also reduced levels of LDH, CPK, and CK-MB in the serum, indicating reduced myocardial damage. Histological analysis revealed that GV treatment attenuated cardiac hypertrophy and fibrosis, with reduced collagen deposition in the myocardium. Immunohistochemistry analysis showed suppressed expression of TGF-β1 and collagen 1, involved in fibrosis. In conclusion, GV showed potential in improving cardiac function in a rat model of MI-induced heart failure. It alleviated myocardial damage, attenuated cardiac hypertrophy and fibrosis, and suppressed fibrotic markers. Further studies are needed to explore its clinical efficacy and underlying mechanisms in cardiac diseases beyond animal models.