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Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 1997.10a
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    • pp.134-152
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    • 1997
  • Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

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Analysis of Factors Affecting the Length of Stay in Children(Aged 0 to 12) with Injuries: Centering Around the Data from the Korea National Hospital Discharge In-Depth Injury Surveys (어린이(0-12세) 손상환자의 재원일수에 미치는 요인분석: 퇴원손상심층자료를 중심으로)

  • Lee Chae Kyung
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.3
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    • pp.137-143
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    • 2023
  • This study was conducted to analyze factors affecting the length of stay in children with injuries by determining relationships between length of stay and characteristics of children(aged 0 to 12) with injuries. 7,804 patients aged 0 to 12 who participated in the Korea Nation Hospital Discharge In-Depth Injury Surveys, got a diagnosis of sequelae of injuries and of other consequences of external causes(S00-T98), and were discharged between 1 January 2016 and 31 December 2020 were investigated. A frequency analysis, independent samples t-test, and ANOVA were performed. Also, to identify factors affecting the length of stay, a regression analysis was performed. The average length of stay for the patients investigated in this study was 5.5 days. The length of stay for school-age children(aged 7 to 12) and children who had either public or private coverage was higher than that for preschoolers(aged 0 to 6) and children who didn't have public or private coverage, respectively. The length of stay for children admitted to a hospital in a rural area(Jeolla-do or Gyeongsang-do) was higher than that for children admitted to a hospital in a metropolitan area and the length of stay for children admitted to a hospital that had 100-299 hospital beds was relatively long. However, children who first visited a hospital for outpatient care stayed relatively short in hospital and children who had been burned or injured in traffic crashes stayed relatively long in hospital. Children who got a secondary diagnosis and had a principal procedure or who died after being discharged were in hospital for a long time. The findings of this study shall be useful, as they identified characteristics related to the length of stay for Korean children with injuries and factors that determine the length of stay for those children by analyzing the national dataset, or more specifically, the data from the Korea National Hospital Discharge In-Depth Injury Surveys. The risk of child injuries can be easily reduced by taking actions to prevent them and providing safety education programs. The present study has provided essential baseline data for the provision of aggressive care for child injuries and the establishment of a range of policies for child injury prevention.

MicroRNA Profile in the Helicobacter pylori-infected Gastric Epithelial Cells (Helicobacter pylori 감염 위상피세포에서 MicroRNA 발현 변화)

  • Chang Whan Kim;Sung Soo Kim;Tae Ho Kim;Woo Chul Chung;Jae Kwang Kim
    • Journal of Digestive Cancer Research
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    • v.5 no.2
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    • pp.105-112
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    • 2017
  • Background: The expression of miRNAs in response to Helicobacter pylori infection has not been well explored. The aims of this study were to evaluate the H. pylori associated miRNAs in the gastric epithelial cells. Methods: We investigated gastric epithelial cell-line (HS3C) exposed H. pylori over 3 months and AGS cell-line (AGS) exposed H. pylori for 6 hour. After the extraction of miRNA from these cell-lines, microarray and real time PCR were performed to confirm the alteration of expression. Results: All 12 miRNAs chosen for real-time PCR are based on the result of microarray and their potential functions related to H. pylori infection. miR-21, miR-221, miR-222 were upregulated in the H. pylori infected AGS cell for 6 hours and HS3C cells. miR-99b, miR-200b, miR-203b and miR-373 were downregulated in the H. pylori infected AGS cell for 6 hours and HS3C cells. miR-23a, miR-23b, miR-125b, miR-141 and miR-155 were upregulated in HS3C cell line but not in H. pylori infected AGS cell for 6 hours. Conclusion: miR-21, miR-99b, miR-125b, miR-200b, miR-203b, miR-221, miR-222, and miR-373 are supposed to be related with oncogenesis of H. pylori infection. Further studies are needed for the evaluation of the function of these confirmed miRNAs.

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Association Between Psychiatric Medications and Urinary Incontinence (정신과 약물과 요실금의 연관성)

  • Jaejong Lee;SeungYun Lee;Hyeran Ko;Su Im Jin;Young Kyung Moon;Kayoung Song
    • Korean Journal of Psychosomatic Medicine
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    • v.31 no.2
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    • pp.63-71
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    • 2023
  • Urinary incontinence (UI), affecting 3%-11% of males and 25%-45% of females globally, is expected to rise with an aging population. It significantly impacts mental health, causing depression, stress, and reduced quality of life. UI can exacerbate psychiatric conditions, affecting treatment compliance and effectiveness. It is categorized into transient and chronic types. Transient UI, often reversible, is caused by factors summarized in the acronym DIAPPERS: Delirium, Infection, Atrophic urethritis/vaginitis, Psychological disorders, Pharmaceuticals, Excess urine output, Restricted mobility, Stool impaction. Chronic UI includes stress, urge, mixed, overflow, functional, and persistent incontinence. Drug-induced UI, a transient form, is frequently seen in psychiatric treatment. Antipsychotics, antidepressants, and other psychiatric medications can cause UI through various mechanisms like affecting bladder muscle tone, altering nerve reflexes, and inducing other conditions like diabetes or epilepsy. Specific drugs like lithium and valproic acid have also been linked to UI, though mechanisms are not always clear. Managing UI in psychiatric patients requires careful monitoring of urinary symptoms and judicious medication management. If a drug is identified as the cause, options include discontinuing, reducing, or adjusting the dosage. In cases where medication continuation is necessary, additional treatments like desmopressin, oxybutynin, trihexyphenidyl, or amitriptyline may be considered.

Attenuation of Oxidative Stress-Induced HepG2 Cellular Damage by Cirsiumjaponicum Root Extract (HepG2 세포에서 대계 추출물에 의한 산화적 스트레스 유발 세포 손상의 억제)

  • Da Jung Ha;Seohwi Kim;Byunwoo Son;Myungho Jin;Sungwoo Cho;Sang Hoon Hong;Yung Hyun Choi;Sang Eun Park
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1002-1014
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    • 2023
  • The root of Cirsium japonicum var. maackii (Maxim.) has long been used in traditional medicine to prevent the onset and progression of various diseases and has been reported to exert a wide range of physiological effects, including antioxidant activity. However, research on its effects on hepatocytes remains scarce. This study used the human hepatocellular carcinoma HepG2 cell line to investigate the antioxidant activity of ethanol extract of C. japonicum root (EECJ) on hepatocytes. Hydrogen peroxide (H2O2) was used to mimic oxidative stress. The results showed that EECJ significantly reverted the decrease in cell viability and suppressed the release of lactate dehydrogenase in HepG2 cells treated with H2O2. Moreover, an analysis of changes in cell morphology, flow cytometry, and microtubule-associated protein light chain 3 (LC3) expression showed that EECJ significantly inhibited HepG2 cell autophagy induced by H2O2. Furthermore, it attenuated H2O2-induced apoptosis and cell cycle disruption by blocking intracellular reactive oxygen species and mitochondrial superoxide production, indicating strong antioxidant activity. EECJ also restored the decreased levels of intracellular glutathione (GSH) and enhanced the expression and activity of superoxide dismutase and GSH peroxidase in H2O2-treated HepG2 cells. Although an analysis of the components contained in EECJ and in vivo validation using animal models are needed, these findings indicate that EECJ is a promising candidate for the prevention and treatment of oxidative stress-induced liver cell damage.

The Role of Ref-1 in the Differentiation Process of Monocytic THP-1 Cells (단핵구세포주 THP-1의 분화과정에서 Ref-1의 역할)

  • Da Sol Kim;Kang Mi Kim;Koanhoi Kim;Young Chul Park
    • Journal of Life Science
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    • v.34 no.4
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    • pp.271-278
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    • 2024
  • Redox factor (Ref)-1, a ubiquitously expressed protein, acts as a modulator of redox-sensitive tran- scription factors and as an endonuclease in the repair pathway of damaged DNA. However, the function of Ref-1 in the differentiation of monocytes into macrophages has not been defined. In this study, we investigated the effects of Ref-1 on the monocyte differentiation process using the human monocytic cell line THP-1. The differentiation agent PMA increased cell adhesion over time and showed a sig- nificant increase in phagocytic function but decreased the intracellular amount of Ref-1. Ref-1 inhibitor E3330 and Ref-1 knockdown using the siRNA technique reduced cell adhesion and the expression of differentiation markers, such as CD14, ICAM-1, and CD11b, by PMA stimulation. This means that the role of Ref-1 is absolutely necessary in the initial process of differentiating THP-1 cells stimulated by PMA. Next, the distribution of Ref-1 was examined in the cytoplasm and nucleus of THP-1 cells stimulated with PMA. Surprisingly, PMA stimulation resulted in the rapid translocation of Ref-1 to the nucleus. To prove that movement of Ref-1 to the nucleus is required for monocyte differentiation, a Ref-1 vector with the nuclear localization sequence (NLS) deleted was used. As a result, overexpression of ∆NLS Ref-1, which restricted movement to the nucleus, suppressed the expression of differentiation markers and notably reduced phagocytic function in PMA-stimulated THP-1 cells. In conclusion, these data suggest that the differentiation of monocytic THP-1 cells requires Ref-1 nuclear translocation during the initial process of biochemical events following stimulation from PMA.

L-AHG-mediated Suppression of M1 Polarization and Pro-inflammatory Signaling Pathways in LPS-stimulated RAW264.7 Macrophages (LPS에 의해 자극된 RAW264.7 대식세포에서 L-AHG에 의한 M1 분극화 및 친염증 신호 경로의 억제)

  • Won Young Jang;Shin Young Park;Ki Youn Kim;Do Youn Jun;Young-Seuk Bae;Young Ho Kim
    • Journal of Life Science
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    • v.34 no.7
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    • pp.443-452
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    • 2024
  • This study aimed to examine the influence of 3,6-anhydroxygalactose (L-AHG) on the pro-inflammatory M1 polarization and pro-inflammatory responses observed in the RAW264.7 mouse macrophage cell line following stimulation with lipopolysaccharides (LPS). L-AHG exhibited a significant and dose-dependent inhibition of inducible nitric oxide synthase (iNOS) expression, a hallmark of M1 polarization, and subsequent NO production in LPS-stimulated RAW264.7 cells. Furthermore, the LPS-induced upregulation of cyclooxygenase-2 (COX-2), which drives the production of prostaglandin E2, an inflammatory mediator, was also inhibited by L-AHG. L-AHG did not affect the LPS-triggered Toll-like receptor 4 (TLR4)-mediated pro-inflammatory signaling pathway, which culminated in the activation of transforming growth factor-β-activated kinase 1 (TAK1). However, it was observed to inhibit the generation of reactive oxugen species (ROS) in a dose-dependent manner, as well as the TAK1-driven activation of JNK and p38 MAPK. Given that the active p38 MAPK is known to contribute to the assembly of active nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which catalyzes the intracellular generation of pro-inflammatory ROS in LPS-stimulated macrophages, the dose-dependent reduction in the LPS-induced ROS generation by L-AHG may be mainly due to the prevention of TAK1-driven activation of p38 MAPK. Together, these results demonstrate that the L-AHG-mediated inhibition of the TAK1-JNK/p38 MAPK activation phase of the pro-inflammatory signaling pathway in LPS-stimulated RAW264.7 cells by L-AHG represents a promising mechanism for suppressing M1 polarization and pro-inflammatory responses in macrophages.

Establishment of Test Conditions and Interlaboratory Comparison Study of Neuro-2a Assay for Saxitoxin Detection (Saxitoxin 검출을 위한 Neuro-2a 시험법 조건 확립 및 실험실 간 변동성 비교 연구)

  • Youngjin Kim;Jooree Seo;Jun Kim;Jeong-In Park;Jong Hee Kim;Hyun Park;Young-Seok Han;Youn-Jung Kim
    • Journal of Marine Life Science
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    • v.9 no.1
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    • pp.9-21
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    • 2024
  • Paralytic shellfish poisoning (PSP) including Saxitoxin (STX) is caused by harmful algae, and poisoning occurs when the contaminated seafood is consumed. The mouse bioassay (MBA), a standard test method for detecting PSP, is being sanctioned in many countries due to its low detection limit and the animal concerns. An alternative to the MBA is the Neuro-2a cell-based assay. This study aimed to establish various test conditions for Neuro-2a assay, including cell density, culture conditions, and STX treatment conditions, to suit the domestic laboratory environment. As a result, the initial cell density was set to 40,000 cells/well and the incubation time to 24 hours. Additionally, the concentration of Ouabain and Veratridine (O/V) was set to 500/50 μM, at which most cells died. In this study, we identified eight concentrations of STX, ranging from 368 to 47,056 fg/μl, which produced an S-shaped dose-response curve when treated with O/V. Through inter-laboratory variability comparison of the Neuro-2a assay, we established five Quality Control Criteria to verify the appropriateness of the experiments and six Data Criteria (Top and Bottom OD, EC50, EC20, Hill slop, and R2 of graph) to determine the reliability of the experimental data. The Neuro-2a assay conducted under the established conditions showed an EC50 value of approximately 1,800~3,500 fg/μl. The intra- & inter-lab variability comparison results showed that the coefficients of variation (CVs) for the Quality Control and Data values ranged from 1.98% to 29.15%, confirming the reproducibility of the experiments. This study presented Quality Control Criteria and Data Criteria to assess the appropriateness of the experiments and confirmed the excellent repeatability and reproducibility of the Neuro-2a assay. To apply the Neuro-2a assay as an alternative method for detecting PSP in domestic seafood, it is essential to establish a toxin extraction method from seafood and toxin quantification methods, and perform correlation analysis with MBA and instrumental analysis methods.

In Vitro Anti-aging and Hair Follicle Dermal Papilla Cells Activation Effects of Usnea diffracta Vain Extract (송라 추출물의 세포 수준에서 항노화 및 모유두세포 활성화 효과)

  • Min Jeong Kim;Won Yeoung Choi;Hyun Woo Shim;Eun Jin Shin;Jung No Lee;Sung Min Park;Hwa Sun Ryu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.1
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    • pp.37-48
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    • 2024
  • Songla (Usnea diffracta Vain.) is one of the lichens belonging to the genus Usnea, and pharmacological activities such as antioxidant, antimicrobial, anti-inflammatory, anti-tumor and cardiovascular protection have been reported in previous studies, but its efficacy in skin and hair is not well known. In this study, the effect of Usnea diffracta extract (UDE) on anti-aging and dermal papilla cell proliferation was verified in vitro. As a result of the experiment, it was confirmed that the UDE significantly reduced the expression of MMP-1 and the activity of MAPKs (ERK, p38, JNK) and AP-1 (c-Fos, c-Jun), which were increased by UVA in HDFn. In addition, the UDE significantly increased the proliferation of HFDPC and significantly increased the mRNA expression of VEGF and KGF, which are hair growth factors. Accordingly, the phosphorylation of ERK/CREB involved in hair proliferation and expression of growth factors was increased in a concentration-dependent manner. The main component represented by the main peak was separated and purified using Prep LC by concentrating the UDE, which was confirmed as diffractaic acid through NMR and Mess analysis. Isolated diffractaic acid significantly reduced the expression of MMP-1 increased by UVA in HDFn and increased the proliferation of HFDPC in a concentration-dependent manner. The result suggest that UDE proved its usability as a natural cosmetic material with anti-aging and dermal papilla cell activation effects.

Involvement of TLR4-JNK/NF-κB signaling pathway in RAW264.7 cell activation of Protaetia brevitarsis seulensis larvae extracts (흰점박이꽃무지 유충 추출물의 RAW264.7 세포 활성화에서 TLR4-JNK/NF-κB 신호전달 경로의 관여)

  • Ju-Hwi Park;Jongbeom Chae;Joon Ha Lee;Dongyup Hahn;Ju-Ock Nam
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.447-454
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    • 2023
  • In the environment in which humans live, there are various antigens that invade the human body and interfere with humans leading a healthy life, so the immune system recognizes the antigen then removes them through a complex mechanism. Macrophages are widely distributed immune cells involved in the innate immune system, and produce various immune modulators such as inducible nitric oxide synthase-induced nitric oxide, cyclooxygenase-2 induced prostaglandin E2 and proinflammatory cytokines such as tumor necrosis factor-alpha. On the other hand, Protaetia brevitarsis seulensis larvae are a type of edible insect that have emerged as an alternative to the future food supply problem. The immuno-modulatory effect through the activation of murine macrophage RAW264.7 cell via mitogen-activated protein kinases (MAPKs)/nuclear factor-kappa B (NF-κB) signaling pathways has been reported. Based on this report, in this study, we confirmed how the expression of immune modulators induced by Protaetia brevitarsis seulensis larvae extracts in RAW264.7 cells was changed by treatment with pharmacological inhibitors of toll-like receptor 4 (TLR4), MAPKs and NF-κB signaling pathways. As a result, reduction of immune modulators was confirmed in the c-Jun N-terminal kinase (JNK) inhibitor treatment group and NF-κB inhibitor treatment group among the Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell. Furthermore, in the TLR4 inhibitor-treated group, decreases in phosphorylation of JNK and NF-κB factors were confirmed in Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell, as well as decreases in immune modulators. This results suggest that Protaetia brevitarsis seulensis larvae activates RAW264.7 cells by the engagement of TLR4-JNK/NF-κB signaling pathway.