• Title/Summary/Keyword: Molecular-level understanding

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Molecular detection of Kudoa septempunctata (Myxozoa: Multivalvulida) in sea water and marine invertebrates

  • Paari, Alagesan;Jeon, Chan-Hyeok;Choi, Hye-Sung;Jung, Sung-Hee;Kim, Jeong-Ho
    • Fisheries and Aquatic Sciences
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    • v.20 no.8
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    • pp.16.1-16.8
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    • 2017
  • The exportation of cultured olive flounder (Paralichthys olivaceus) in Korea has been recently decreasing due to the infections with a myxozoan parasite Kudoa septempunctata, and there is a strong demand for strict food safety management because the food poisoning associated with consumption of raw olive flounder harbouring K. septempunctata has been frequently reported in Japan. The life cycle and infection dynamics of K. septempunctata in aquatic environment are currently unknown, which hamper establishment of effective control methods. We investigated sea water and marine invertebrates collected from olive flounder farms for detecting K. septempunctata by DNA-based analysis, to elucidate infection dynamics of K. septempunctata in aquaculture farms. In addition, live marine polychaetes were collected and maintained in well plates to find any possible actinosporean state of K. septempunctata. The level of K. septempunctata DNA in rearing water fluctuated during the sampling period but the DNA was not detected in summer (June-July in farm A and August in farm B). K. septempunctata DNA was also detected in the polychaetes Naineris laevigata intestinal samples, showing decreased pattern of 40 to 0%. No actinosporean stage of K. septempunctata was observed in the polychaetes by microscopy. The absence of K. septempunctata DNA in rearing water of fish farm and the polychaetes N. laevigata intestinal samples during late spring and early summer indicate that the infection may not occur during this period. N. laevigata was suspected as the possible alternate invertebrate host of K. septempunctata, but the actinosporean stage was not found by well plate method and further studies will be necessary. This research provides important baseline information for understanding the infection dynamics of K. septempunctata in olive flounder farms and further establishment of control strategies.

Comparison of Anti-cancer Potentials of Water Extracts of Bigihwan, Daechilgithang and Mokwhyangbinranghwan in Human Hepatocellular Carcinoma Cells (인체 간암세포에서 비기환(肥氣丸), 대칠기탕(大七氣湯) 및 목향빈랑환(木香檳榔丸) 열수 추출물의 항암 활성 비교)

  • Kim, Min Yeong;Lee, Hyesook;Hong, Su Hyun;Park, Cheol;Choi, Yung Hyun
    • Herbal Formula Science
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    • v.28 no.1
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    • pp.15-27
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    • 2020
  • Objectives : We selected three herb-combined remedies, Bigihwan (BGH), Daechilgitang (DCGT) and Mokwhyangbinranghwan (MHBRH), through Donguibogam text-mining analysis, and evaluated their anti-cancer effects on human hepatocellular carcinoma Hep3B cells. Methods : Cytotoxicity was assessed by an MTT assay. Apoptosis rate, autophagy and ROS level were detected by flow cytometry. The autophagy was also observed by Cyto-ID staining fluorescence microscopy. The expression of autophagy, mitophagy and pexophagy regulatory proteins was detected by Western blot analysis. Results : BGH showed the strongest effect among the three prescriptions in inhibiting Hep3B cell viability, which was associated with the induction of apoptosis and autophagy. Autophagy blockers improved cell viability and reduced apoptosis after BGH and DCGT treatments, indicating that autophagy by these prescriptions enhanced Hep3B cells against their cytotoxicity. However, MHBRH enhanced the reduction of cell viability and apoptosis by autophagy blockers. Induction of autophagy by BGH treatment was associated with mitophagy due to mitochondrial dysfunction than DCGT and MHBRH-treated cells. In addition, induction of apoptosis by BGH treatment was ROS-dependent and showed the possibility of pexophagy involvement. Conclusion : Although further studies need to be conducted to study the efficacy and mechanism of accurate anticancer activity, the present results will serve as important sources of understanding the mechanism of action of herbal remedies prescribed for liver disease as documented in Donguibogam.

TNF-induced genes and Proteins

  • 이태호
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.11a
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    • pp.17-20
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    • 1993
  • As a step toward a more complete understanding of the molecular actions of TNF, we prepared a cDNA library from TNF-treated human FS-4 fibroblasts and used differential hybridization to identify cDNA clones corresponding to mRNAs enriched in TNF-treated eells. In Quiescent FS-4 cells n induces an increase in the level of some mRNAs within 20 to 30 min. Some of these immediate-early response mRNAs are elevated only transiently for about 30 to 120 min, e. g., c-fos and c-myc (Lin and Vilcek,1987) or the transcription factor IRF-1 (Fujita et al.1989). Such immediate-early gene products may be important for the activation of other genes, but their transient induction suggests that they are not the actual effector molecules responsible for the phenotypic changes induced by TNF. We chose a 3-h incubation with W because we were seeking cDNAs corresponding to messages that are more stably elevated after TNF treatment. Indeed, the results shown in Figure 8 and 9 indicate that all of the mRNAs corresponding to the eight TSG cDNAs isolated remained significantly elevated after 16h of continuous treatment with TNF, and their kinetics of induction were clearly different from those of the immediate-early response mRNAs such as c-fos, c-myc or IRF-1. Nevertheless, only the induction of TSG-21 (collagenase) and TSG-27 (stromelysin) nNAs was completely inhibited by cycloheximide and the induction of TSG-37 (metallothionein-II) was reduced in the presence of this inhibitor of protein synthesis. Induction of the other five TSG mRNAs by TNF was completelyresistant to cycloheximide, suggest ins that no protein intermediate is needed for the upregulation of these mRNAs.

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Characterization of Novel Trichoderma asperellum Isolates to Select Effective Biocontrol Agents Against Tomato Fusarium Wilt

  • El_Komy, Mahmoud H.;Saleh, Amgad A.;Eranthodi, Anas;Molan, Younes Y.
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.50-60
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    • 2015
  • The use of novel isolates of Trichoderma with efficient antagonistic capacity against Fusarium oxysporum f. sp. lycopersici (FOL) is a promising alternative strategy to pesticides for tomato wilt management. We evaluated the antagonistic activity of 30 isolates of T. asperellum against 4 different isolates of FOL. The production of extracellular cell wall degrading enzymes of the antagonistic isolates was also measured. The random amplified polymorphic DNA (RAPD) method was applied to assess the genetic variability among the T. asperellum isolates. All of the T. asperellum isolates significantly reduced the mycelial growth of FOL isolates but the amount of growth reduction varied significantly as well. There was a correlation between the antagonistic capacity of T. asperellum isolates towards FOL and their lytic enzyme production. Isolates showing high levels of chitinase and ${\beta}$-1,3-glucanase activities strongly inhibited the growth of FOL isolates. RAPD analysis showed a high level of genetic variation among T. asperellum isolates. The UPGMA dendrogram revealed that T. asperellum isolates could not be grouped by their antagonistic behavior or lytic enzymes production. Six isolates of T. asperellum were highly antagonistic towards FOL and potentially could be used in commercial agriculture to control tomato wilt. Our results are consistent with the conclusion that understanding the genetic variation within Trichoderma isolates and their biochemical capabilities are required for the selection of effective indigenous fungal strains for the use as biocontrol agents.

Conifer Somatic Embryogenesis : New Knowledge in Plant Biology and Breakthrough in Tree Biotechnology (침엽수(針葉樹) 체세포(體細胞) 배발생(胚發生)을 이용(利用)한 임목생물공학(林木生物工學))

  • Bozhkov, P.V.;Park, Y.G.
    • Journal of Korean Society of Forest Science
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    • v.85 no.4
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    • pp.667-679
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    • 1996
  • Clonal forestry and reforestation programmes are especially interested now in development and application of controllable biotechnological systems based on the production of conifer somatic embryos in bioreactors with their following drilling and/or storage in the form of "artificial seeds". Modern achievements in conifer somatic embryogenesis has guided the development not only of biotechnological systems in forestry, but also of basic research in conifer embryology, cell and molecular biology. At the present time, the level of development of applied research on conifer somatic embryogenesis is well ahead our understanding of this complex phenomenon. The "bottleneck" situation in relation between basic and applied sciences will eventually lead to the appearance of "weak points" in biotechnological systems. In the present review, the major advances and the most pressing problems in the application of conifer somatic embryogenesis both to forest biotechnology and to basic research are in the focus of attention.

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The Effects of Problem Solving Strategy and Paired Think-Aloud Problem Solving on High School Students' Chemistry Problem Solving (문제 해결 전략과 해결자.청취자 활동이 고등학생의 화학 문제 해결에 미치는 효과)

  • Jeon, Kyung-Moon;Noh, Tae-Hee
    • Journal of The Korean Association For Science Education
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    • v.21 no.2
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    • pp.289-298
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    • 2001
  • The effect of the instructional approach that asked students to check their problem-solving processes through a paired think-aloud problem solving after presenting molecular-level pictures and a four stage-problem solving strategy was investigated. Four high school classes (N = 191) were randomly assigned to St group (using Strategy individually), SL group (Solver Listener), St-SL group (using Strategy-Solver Listener), and control group. Although the test scores of the St-SL group on strategy performing ability were significantly higher than those of the control group, there was not significant difference for the scores in the multiple-choice algorithmic problems. Regarding the subcategories of strategy performing ability test, students' ability of understanding given of problems and deriving the proper physical quantity was improved, but their ability of setting up subgoals and reviewing their solving process was very low. The preference to the strategy of the St-SL group was more positive than that of the St group.

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Comparative N-Linked Glycan Analysis of Wild-Type and α1,3-Galactosyltransferase Gene Knock-Out Pig Fibroblasts Using Mass Spectrometry Approaches

  • Park, Hae-Min;Kim, Yoon-Woo;Kim, Kyoung-Jin;Kim, Young June;Yang, Yung-Hun;Jin, Jang Mi;Kim, Young Hwan;Kim, Byung-Gee;Shim, Hosup;Kim, Yun-Gon
    • Molecules and Cells
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    • v.38 no.1
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    • pp.65-74
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    • 2015
  • Carbohydrate antigens expressed on pig cells are considered to be major barriers in pig-to-human xenotransplantation. Even after ${\alpha}1,3$-galactosyltransferase gene knock-out (GalT-KO) pigs are generated, potential non-Gal antigens are still existed. However, to the best of our knowledge there is no extensive study analyzing N-glycans expressed on the GalT-KO pig tissues or cells. Here, we identified and quantified totally 47 N-glycans from wild-type (WT) and GalT-KO pig fibroblasts using mass spectrometry. First, our results confirmed the absence of galactose-alpha-1,3-galactose (${\alpha}$-Gal) residue in the GalT-KO pig cells. Interestingly, we showed that the level of overall fucosylated N-glycans from GalT-KO pig fibroblasts is much higher than from WT pig fibroblasts. Moreover, the relative quantity of the N-glycolylneuraminic acid (NeuGc) antigen is slightly higher in the GalT-KO pigs. Thus, this study will contribute to a better understanding of cellular glycan alterations on GalT-KO pigs for successful xenotransplantation.

Identification of Proteomic Components Associated with Resistance to Fusarium Head Blight in Rye

  • Perlikowski, Dawid;Wisniewska, Halina;Goral, Tomasz;Ochodzki, Piotr;Majka, Maciej;Pawlowicz, Izabela;Belter, Jolanta;Kosmala, Arkadiusz
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.313-320
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    • 2019
  • Rye was used here to dissect molecular mechanisms of resistance to Fusarium head blight (FHB) and to go deeper with our understanding of that process in cereals. F. culmorum-damaged kernels of two lines different in their potential of resistance to FHB were analyzed using two-dimensional gel electrophoresis and mass spectrometry to identify resistance markers. The proteome profiling was accompanied by measurements of ${\alpha}-$ and ${\beta}-amylase$ activities and mycotoxin content. The proteomic studies indicated a total of 18 spots with clear differences in protein abundance between the more resistant and more susceptible rye lines after infection. Eight proteins were involved in carbohydrate metabolism of which six proteins showed a significantly higher abundance in the resistant line. The other proteins recognized here were involved in stress response and redox homeostasis. Three remaining proteins were associated with protease inhibition/resistance and lignin biosynthesis, revealing higher accumulation levels in the susceptible rye line. After inoculation, the activities of ${\alpha}-$ and ${\beta}-amylases$, higher in the susceptible line, were probably responsible for a higher level of starch decomposition after infection and a higher susceptibility to FHB. The presented results could be a good reference for further research to improve crop resistance to FHB.

Screening assay for tomato plants resistant to Fusarium oxysporum f. sp. lycopersici race 2 using the expression of the avr2 gene as a selection marker

  • Kim, Mi-Reu;Lee, Jeong Jin;Min, Jiyoung;Kim, Sun Ha;Kim, Dae-Gyu;Oh, Sang-Keun
    • Korean Journal of Agricultural Science
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    • v.48 no.1
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    • pp.151-161
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    • 2021
  • Fusarium wilt disease of tomato plants caused by Fusarium oxysporum f.sp. lycopersici (FOL race2) is one of the most important diseases of tomatoes worldwide. In the competition between tomato and FOL, the FOL can win by overcoming the immune system of tomato plants. Resistant interaction between the FOL race2 and tomato plants is controlled by avirulence genes (AVR2) in FOL and the corresponding resistance genes (I2) in tomato plants. In this study, 7 FOL isolates (KACC) were used to test their pathogenicity, and FOL race2 was selected because it is a broad problem in Korea. The Fol40044 isolates showed the most severe pathogenicity, and the avr2 gene was also isolated and identified. Moreover, to select resistance, 20 tomato varieties were inoculated with the Fol40044, and the degree of pathogenicity was evaluated by analyzing the expression of the avr2 gene. As a result, three resistant tomato varieties (PCNUF73, PCNUF101, PCNUF113) were selected, and the expression of the avr2 gene was much lower than that of the control Heinz cultivar. This result shows that the screening assay is very efficient when the avr2 gene is used as a marker to evaluate the expression level when selecting varieties resistant to tomato wilt disease. Based on these results, it is possible to isolate the I2 gene, which exhibits resistance and molecular biological interactions with the AVR2 gene from the three tomato-resistant varieties. The I2 gene provides breeders more opportunities for Fusarium disease resistance and may contribute to our understanding of their interactions with the FOL and host plant.

Semen evaluation: methodological advancements in sperm quality-specific fertility assessment - A review

  • Tanga, Bereket Molla;Qamar, Ahmad Yar;Raza, Sanan;Bang, Seonggyu;Fang, Xun;Yoon, Kiyoung;Cho, Jongki
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1253-1270
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    • 2021
  • Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.