• Title/Summary/Keyword: RNA polymerase II

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Species Diversity of Endophytic Fungi Isolated from Taxus cuspidata Inhabiting Mt. Hallasan, Korea

  • Jae-Eui Cha;Hyeok Park;Ahn-Heum Eom
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.419-428
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    • 2023
  • Several endophytic fungal strains were isolated from Taxus cuspidata and identified by molecular analysis of the internal transcribed spacer and RNA polymerase II second largest subunit. This study aimed to determine the relative abundance and compare the species diversity of endophytic fungal communities within needle leaves and twigs. We identified a total of 49 endophytic fungal species. Notably, two species, Trichoderma dingleyae and Xylaria cubensis, were discovered to be previously unrecorded in Korea. The fungal communities in both plant tissues demonstrated distinct species composition. Differences were observed in the relative abundance and species diversity index between needle leaves and twigs. Our findings suggest that the host plant tissues influence the species diversity of endophytic fungal communities.

Upregulation of Renal Renin-Angiotensin System in Rats with Adriamycin-Induced Nephrosis

  • Kim, Soo-Wan;Lee, Jong-Un;Han, Sang-Woong;Ryu, Jun-Ho;Oh, Yoon-Wha;Kim, Nam-Ho;Choi, Ki-Chul;Kim, Ho-Jung
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.2
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    • pp.127-130
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    • 2002
  • The present study was aimed to investigate whether the adriamycin-induced nephrosis is associated with an altered regulation of local renin-angiotensin system (RAS) in the kidney. Rats were subjected to a single injection of adriamycin (2 mg/kg body weight, IV) and kept for 6 weeks to allow the development of nephrosis. They were then divided into two groups, and supplied with and without cilazapril, an angiotensin converting enzyme (ACE) inhibitor, in drinking water (100 mg/l) for additional 6 weeks. Another group without adriamycin-treatment served as control. The mRNA expression of renin, ACE, type 1 and type 2 angiotensin II receptors (AT1R, AT2R), and transforming growth factor $(TGF)-{\beta}1$ was determined in the cortex of the kidney by reverse transcription-polymerase chain reaction. Adriamycin treatment resulted in heavy proteinuria. Accordingly, the mRNA expression of renin, ACE, and AT1R was increased in the renal cortex, while that of AT2R was decreased. Co-treatment with cilazapril attenuated the degree of proteinuria. While not affecting the altered expression of renin, cilazapril decreased the expression of ACE to the control level. Cilazapril further increased the expression of AT1R, while it restored the decreased expression of AT2R. The expression of $(TGF)-{\beta}1$ was increased by the treatment with adriamycin, which was abolished by cilazapril. An altered expression of local RAS components may be causally related with the development of adriamycin-induced nephrosis, in which AT1R is for and AT2R is against the development of nephrosis.

Dronedarone Attenuates Ang II-Induced Myocardial Hypertrophy Through Regulating SIRT1/FOXO3/PKIA Axis

  • Cheng Chen;Song Hu;Heng-Jing Hu;Zhi-Xuan Liu;Xin-Teng Wu;Tao Zou;Hua Su
    • Korean Circulation Journal
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    • v.54 no.4
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    • pp.172-186
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    • 2024
  • Background and Objectives: Long-term pathological myocardial hypertrophy (MH) seriously affects the normal function of the heart. Dronedarone was reported to attenuate left ventricular hypertrophy of mice. However, the molecular regulatory mechanism of dronedarone in MH is unclear. Methods: Angiotensin II (Ang II) was used to induce cell hypertrophy of H9C2 cells. Transverse aortic constriction (TAC) surgery was performed to establish a rat model of MH. Cell size was evaluated using crystal violet staining and rhodamine phalloidin staining. Reverse transcription quantitative polymerase chain reaction and western blot were performed to detect the mRNA and protein expressions of genes. JASPAR and luciferase activity were conducted to predict and validate interaction between forkhead box O3 (FOXO3) and protein kinase inhibitor alpha (PKIA) promoter. Results: Ang II treatment induced cell hypertrophy and inhibited sirtuin 1 (SIRT1) expression, which were reversed by dronedarone. SIRT1 overexpression or PKIA overexpression enhanced dronedarone-mediated suppression of cell hypertrophy in Ang II-induced H9C2 cells. Mechanistically, SIRT1 elevated FOXO3 expression through SIRT1- mediated deacetylation of FOXO3 and FOXO3 upregulated PKIA expression through interacting with PKIA promoter. Moreover, SIRT1 silencing compromised dronedarone-mediated suppression of cell hypertrophy, while PKIA upregulation abolished the influences of SIRT1 silencing. More importantly, dronedarone improved TAC surgery-induced MH and impairment of cardiac function of rats via affecting SIRT1/FOXO3/PKIA axis. Conclusions: Dronedarone alleviated MH through mediating SIRT1/FOXO3/PKIA axis, which provide more evidences for dronedarone against MH.

Immune-Enhancing Effects of Polysaccharides Isolated from Ascidian (Halocynthia roretzi) Tunic (우렁쉥이(Halocynthia roretzi) 껍질로부터 분리된 다당류의 면역증강 효과)

  • Lee Dae-Hoon;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.673-680
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    • 2015
  • In this study, the immune-enhancing effects of purified polysaccharides from ascidian (Halocynthia roretzi) tunic were investigated. Crude polysaccharides (AP) were isolated by enzyme extraction (neutrase, $60^{\circ}C$, 15h), ethanol precipitation, and lyophilization. In addition, crude polysaccharides were further fractionated into unabsorbed fractions (APF-I, fraction No. 11~17) and absorbed fractions (APF-II, fraction No. 22~37) by DEAE-sepharose CL-6B column chromatography in order to isolate immune regulating polysaccharide. The major constituents in APF-I and APF-II were total sugar (66.62% and 27.03%), uronic acid (47.53% and 15.87%), hexosamine (16.62% and 46.79%), and protein (2.43% and 4.94%), respectively. APF-I increased production of nitric oxide (NO) and cytokines, such as tumor necrosis factor-alpha (TNF-${\alpha}$) and interleukin (IL)-6 in a dose-dependent manner. The mRNA expression levels of inducible NO synthetase, cyclooxygenase-2, TNF-${\alpha}$, and IL-6 were markedly increased as determined by polymerase chain reaction analysis. The above data led us to conclude that macrophage activation of purified polysaccharides was higher than that of crude polysaccharides. The polysaccharides isolated from ascidian tunic investigated herein are useful as natural immune enhancing agents.

Characterization of Fusarium solani Causing Fusarium Root Rot of Lisianthus in Korea (꽃도라지 뿌리썩음병을 일으키는 Fusarium solani 의 특성)

  • Choi, Hyo-Won;Hong, Sung Kee;Lee, Young Kee;Kim, Jeomsoon;Lee, Jae Guem;Kim, Hyo Won;Kang, Eun Hye;Lee, Eun Hyeong
    • The Korean Journal of Mycology
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    • v.45 no.1
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    • pp.74-82
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    • 2017
  • Lisianthus (Eustoma grandiflorum) is a flowering ornamental plant used widely in Korea. In 2015, wilting, damping-off, stunting, and root rot symptoms were observed in lisianthus plants of Yeoju and Gimhae, Korea. Affected seedlings appeared yellow and showed poor development of root systems in the field and in nursery boxes. Furthermore, affected plants were yellow, stunted, and died at approximately 2-3 months after transplanting. Fusarium species were consistently isolated from the basal stems of diseased plants. Nine isolates were identified as Fusarium solani based on morphological characteristics. Macroconidia of isolates were relatively wide, straight-to-slightly curved, and microconidia formed in false heads on long monophialides. Abundant chlamydospores were produced at the middle or tips of hyphae. To confirm this identification, a molecular analysis of the translation elongation factor 1 alpha (TEF) and RNA polymerase II subunit (RPB2) genes was conducted. The sequences of TEF and RPB2 showed 99.2-99.9% and 98.0-98.1% similarity, respectively, to those of reference F. solani strains in NCBI GenBank. Pathogenicity was tested using root dipping inoculation of healthy lisianthus seedlings. Symptoms were observed within 7 days of inoculation only in inoculated plants. This is the first report of F. solani causing Fusarium root rot on lisianthus in Korea.

Resveratrol Upregulates p21, Cell Cycle Regulator, in Gaucher Disease Cells (Gaucher병에서 resveratrol의 세포주기 조절자 p21을 통한 세포보호 효과 연구)

  • Kim, Dong-Hyun;Heo, Tae-Hwe;Kim, June-Bum;Kim, Sung-Jo
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1281-1286
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    • 2010
  • Gaucher disease (GD) is caused by glucocerebrosidase functional deficiency and the most prevalent lysosomal storage disorder (LSD), with an incidence of about 1 in 20,000 new births. Resveratrol, one kind of phytoalexin, is a produced naturally by several plants and has anti-tumor, anti-aging, anti-inflammatory and neuro-protective effects. In this paper we provide the cellular protective effect of resveratrol in both type I and type II Gaucher disease cells. Resveratrol treatment did not show any significant change in the p21 and p53 mRNA expression level, however expression level of the p21 protein, a cell cycle arrest factor, shows significant increment in both types of Gaucher disease cells. These cell cycle arrest patterns were confirmed by both MTT assay measurement and microscopy detection. In comparison, expression level of poly ADP ribose polymerase (PARP), an apoptosis indicator protein, was significantly decreased in both type I and II Gaucher disease cells after treatment with resveratrol. This result indicates that resveratrol relievescellular apoptotic stress fromtype I and II Gaucher disease cells. Therefore, we demonstrate that resveratrol inhibits cell proliferation via p21 activity and activates cellular repair systems for Gaucher disease cells. Our results provide at least one of the molecular mechanisms of Gaucher disease and may allow the verification of potential drug targets for therapeutic trials.

Healing Effect of Cordyceps Militaris Extract Complex on Collagen II-Induced Arthritis Rats (콜라겐 유도 관절염 동물 모델에 대한 동충하초 복합추출물의 치료 효과)

  • Oh, Seung-Joon;Lee, Eun-Jung
    • Journal of Korean Medicine Rehabilitation
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    • v.32 no.3
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    • pp.1-11
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    • 2022
  • Objectives This study was designed to evaluate the healing effect of Cordyceps Militaris (CM) on collagen II-induced arthritis rats. Methods Sprague-Dawley rats were randomly divided into 6 groups (normal, control, positive control, CM with low/medium/high dosage each). Type II collagen mixed with complete Freund's adjuvant (with 1:1 v/v) was injected subcutaneously, and the mixture was injected in a same manner one week after the first injection to boost arthritis. Arthritis index, paw thickness and von Frey test were conducted to observe physical changes. hematoxylin and eosin (H&E) staining was performed to observe knee cartilage. The levels of messenger RNA (mRNA) expressions of interleukin (IL)-1𝛽, IL-6, tumor necrosis factor-alpha (TNF-𝛼) in spleen were assessed by real-time polymerase chain reaction. Results Rheumatoid arthritis is an autoimmune disease that occurs on multiple joints and can lead to temporary shape change of bones or organ failure in severe cases. Here, we aimed to determine the effect of CM extract on rheumatoid arthritis by measuring paw thickness, arthritis index, conducting von Frey test and H&E staining, and evaluating the level of IL-1𝛽, IL-6, TNF-𝛼. As a result, paw thickness, arthritis index significantly decreased in low concentration group, hind leg became less sensitive in all expermental groups. Also, histological analysis showed that the damage of knee cartilage was prevented in all experimental groups. The level of mRNA of IL-1𝛽, IL-6, and TNF-𝛼 in spleen was analyzed to decide the effectiveness of CM extract. IL-1𝛽 did not show significant change, but IL-6 and TNF-𝛼 showed significant decrease in at least one of the experimental groups. Conclusions CM showed protective effect on knee tissue destruction and improved the physical conditions of the leg involving arthritis. Also, it showed that CM has anti-inflammatory effect on specific cytokines inducing rheumatoid arthritis. In conclusion, this study demonstrated that the therapeutic potential of CM for the treatment rheumatoid arthritis, and set the foundation for the further studies.

Detection of 23S rRNA Mutation Associated with Clarithromycin Resistance in Children with Helicobacter pylori Infection (소아 Helicobacter pylori 감염에서 Clarithromycin 내성과 연관된 23S rRNA의 돌연변이)

  • Ko, Jae Sung;Yang, Hye Ran;Seo, Jeong Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.2
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    • pp.137-142
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    • 2004
  • Purpose: The resistance of H. pylori to clarithromycin is one of the major causes of eradication failure. In H. pylori, clarithromycin resistance is due to point mutation in 23S rRNA. The aims of this study were to investigate the mutation of 23S rRNA and to examine the association of cagA, vacA genotype and clarithromycin resistant genes. Methods: H. pylori DNA was extracted from antral biopsy specimens from 27 children with H. pylori infection. Specific polymerase chain reaction (PCR) assays were used for cagA and vacA. Mutations associated with clarithromycin resistance were detected by using PCR restriction fragment length polymorphism (RFLP) analysis of 23S rRNA gene. Results: A2143G mutation was detected in one case and A2144G in 4, indicating 18.5% were clarithromycin resistant. Among the total of 27, cagA was present in 25 (93%), vacA s1a/m1 in 6 (22%), s1a/m2 in 3 (11%), s1c/m1 in 16 (59%), and s1c/m2 in 1 (4%). All of the 5 clarithromycin resistant strains were cagA (+), among which 2 were s1a/m1 and 2 were s1c/m1. There was no relation between genotypes and clarithromycin resistant genes. Conclusion: Detection of H. pylori resistance to clarithromycin using PCR RFLP from biopsy specimens might be useful for the selection of antibiotics. Clarithromycin resistant genes are not associated with genotypes of cagA and vacA.

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Expression of DNA Methyltransferase Transcripts in The Oocytes and Preimplantation Embryos in Mouse (생쥐 난자와 착상전 초기배아에서 DNA 메틸전이효소 전사물의 발현)

  • 김종월;이양한;강승호;한성원;전일경;김성례;김문규
    • Development and Reproduction
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    • v.2 no.2
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    • pp.197-203
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    • 1998
  • DNA methylation seems to play an important regulatory role in gene expression and cell differentiation during postimplantation embryonic development. However, the significance of DNA methylation which is maintained by the DNA MTase during preimplantation embryonic development, is not fully understood. In order to study the role of DNA methylation in the preimplantation embryos, the expression of DNA MTase transcripts was monitored in the oocytes and preimplantation embryos. The mRNA of DNA MTase was detected in the oocytes and pleimplantation embryos. The relative mRNA levels of DNA MTase were high from the stages of GV-oocytes and pronuclear embryos, and thereafter decreased gradually. By the treatment of $\alpha$-amanitin, it was confirmed that the transcripts presented in pronuclear embryos was derived from the maternal genome. The presence of transcripts of DNA MTase in the oocytes and pronuclear embryos suggests that the maintenance of DNA methylation may be necessary and seems to play an important role in gene expression and cell differentiation during preimplantation embryonic develop-ment in mouse.

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Rapid Detection of Noroviruses in Fecal Samples and Shellfish by Nucleic Acid Sequence-based Amplification

  • Kou Xiaoxia;Wu Qingping;Zhang Jumei;Fan Hongying
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.403-408
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    • 2006
  • The purpose of this study was to determine the efficacy of a nucleic acid sequence-based amplification (NASBA) method of detecting noroviruses in artificially and naturally contaminated shellfish. We used 58 fecal samples that tested positive for noroviruses with electron microscopy (EM) to develop an NASBA assay for these viruses. Oligonucleotide primers targeting the polymerase coding region were used to amplify the viral RNA in an isothermal process that resulted in the accumulation of RNA amplicons. These amplicons were detected by hybridization with digoxigenin-labeled oligonucleotide probes that were highly specific for genogroup I (GI) and genogroup II (GII) of noroviruses. The expected band of 327bp appeared in denaturing agarose gel without any nonspecific band. The specific signal for each amplicon was obtained through Northern blotting in many repeats. All fecal samples of which 46(79.3%) belonged to GII and 12(20.6%) belonged to GI were positive for noroviruses by EM and by NASBA. Target RNA concentrations as low as 5pg/ml were detected in fecal specimens using NASBA. When the assay was applied to artificially contaminated shellfish, the sensitivity to nucleic acid was 100pg/1.5g shellfish tissue. The potential use of this assay was also confirmed in naturally contaminated shellfish collected from different ponds in Guangzhou city of China, of which 24 (18.76%) out of 128 samples were positive for noroviruses; of these, 19 (79.6%) belonged to GII and 5 (20.4%) belonged to GI. The NASBA assay provided a more rapid and efficient way of detecting noroviruses in fecal samples and demonstrated its potential for detecting noroviruses in food and environmental samples with high specificity and sensitivity.