• Title/Summary/Keyword: E6/E7 mRNA

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First Report of Allantophomopsiella pseudotsugae Isolated from Soil in Korea

  • Wajihi, Ally Hassan;Lee, Seung-Yeol;Das, Kallol;Eom, Ahn-Heum;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.29-34
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    • 2019
  • A fungal isolate designated 17E029 was isolated from a soil sample in Jeju, Korea. The strain was similar to other Allantophomopsiella species in its morphological characteristics such as grey mycelia, conidiophore, and conidia sizes. The isolate produced aerial mycelia, which appeared grey on the reverse side of the media surfaces and turned black on the front side of the colonies. The conidiophores emanating from the hyphae were hyaline, grey, aseptate, branched, and $6.7{\sim}9.2{\times}1.8{\sim}2.5{\mu}m$. Conidiogenous cells were ovoid to subcylindrical, discrete, guttulate, and hyaline. Conidia were hyaline, aseptate, smooth, guttulate, oval to subcylindrical, irregular in shape, and $6.0{\sim}7.8{\times}3.0{\sim}3.4{\mu}m$. The strain was confirmed based on phylogenetic analysis of the closest related organism, A. pseudotsugae CBS 288.37, using the partial 28S, internal transcribed spacer rDNA regions, and partial RNA polymerase II second largest subunit locus (RPB2) gene sequences along with its culture characteristics. Therefore, morphological observations and phylogenetic analysis revealed that strain 17E029 is similar to the previously identified A. pseudotsugae. Hence, this species was described as A. pseudotsugae strain 17E029, which is a new record in Korea.

Inhibitory Effect of Brassica rapa in Ovalbumin-Stimulated Experimental Asthmic Mice

  • Bae, Eun-Ah;Min, Seung-Won;Kim, Nam-Jae;Bang, Myung-Ho;Baek, Nam-In;Han, Eun-Joo;Chung, Hae-Gon;Kim, Dong-Hyun
    • Natural Product Sciences
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    • v.13 no.2
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    • pp.114-117
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    • 2007
  • To evaluate antiallergic effect of Brassica rapa Metzg (BR, Family Cruciferae), which is widely used as a food source, its anti-asthmic effect was investigated. BR reduced IgE level in the blood of ovalbumin (OVA)-induced asthmic mice. BR also inhibited IgE and ploinflammatory cytokine IL-6 productions in trachea of OVA-induced mice, but did not inhibit IL-4 production. BR did not inhibit mRNA expression of proinflammatory cytokines IL-6 and IL-4 in IgE-induced RBL-2H3 cells. BR and its isolated components 10-undecenoic acid 2-methoxy methyl ester and galactosyl diglyceride potently inhibited the NO production of RAW264.7cells induced by lipopolysaccharide. These findings suggest that the anti-asthmic effect of BR may be due to the inhibition of the biosynthesis of proinflarnmatory cytokine IL-6 in macrophage and BR can improve IgE-induced allergic disease asthma.

Immune stimulating effects of Astragalus membranaceus and Zanthoxylum schinifolium 1:1 mixture in Raw264.7 cells (Raw264.7 세포에서 황기와 산초 1:1 혼합물의 면역 증진 효과)

  • Il Je Cho;Yeong Eun Yu;Sang Min Lee;Eun Ok Kim;Joon Heum Park;Sea Kwang Ku
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.519-526
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    • 2023
  • Present study explored immunostimulatory effects of Astragalus membranaceus and Zanthoxylum schinifolium 1:1 (w:w) mixture (AZM-1:1) in Raw264.7 cells, mouse macrophage derived cells. Treatment with 100-400 ㎍/mL of AZM-1:1 in Raw264.7 cells significantly increased nitric oxide production in parallel with inducible nitric oxide synthase mRNA expression without affecting cytotoxicity. In addition, AZM-1:1 dose-dependently increased prostaglandin E2 production in conditioned medium along with cyclooxygenase-2 mRNA induction. Moreover, AZM-1:1 induces the transcription of tumor necrosis factor-α, interleukin-1β, interleukin-6, and monocyte chemoattractant protein-1. Immunoblot analyses revealed that AZM-1:1 significantly increased the phosphorylation of mitogen-activated protein kinases, provoked phosphorylation-mediated degradation of inhibitory-κBα, and phosphorylated p65. Furthermore, treatment with AZM-1:1 promoted phagocytosis of Escherichia coli particle labeled with green fluorescence. Taken together, AZM-1:1 may be a promising nutraceutical for stimulation the innate immune system, including macrophages.

In Vitro Immune-Enhancing Activity of Ovotransferrin from Egg White via MAPK Signaling Pathways in RAW 264.7 Macrophages

  • Lee, Jae Hoon;Ahn, Dong Uk;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.38 no.6
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    • pp.1226-1236
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    • 2018
  • Ovotransferrin (OTF) is a well-known protein of the transferrin family with strong iron chelating activity, resulting in its antimicrobial activity. Furthermore, OTF is known to have antioxidant, anticancer, and antihypertensive activities. However, there have been few studies about the immune-enhancing activity of OTF. In current study, we investigated the immune-enhancing activity of OTF using the murine macrophage cells in vitro. The effect of OTF on production of pro-inflammatory mediators and cytokines were determined using Griess assay and quantitative real-time PCR. Using Neutral Red uptake assay, we confirmed the effect of OTF on phagocytic activity of macrophages. Ovotransferrin significantly increased the production of nitric oxide (NO) and secretion of inducible nitric oxide synthase (iNOS) mRNA with no cytotoxic activity. Ovotransferrin (2 mg/mL) stimulated NO production up to $31.9{\pm}3.5{\mu}M$. Ovotransferrin significantly increased the mRNA expression levels of pro-inflammatory cytokines which are tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), Interleukin-$1{\beta}$ (IL-$1{\beta}$), and IL-6: OTF (2 mg/mL) treatment increased the secretion of mRNA for TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 by 22.20-, 37.91-, and 6.17-fold of the negative control, respectively. The phagocytic activity of macrophages was also increased by OTF treatment significantly compared with negative control. Also, OTF treatment increased phosphorylation level of MAPK signaling pathways. These results indicated that OTF has immune-enhancing activity by activating RAW 264.7 macrophages via MAPK pathways.

Expression patterns of TRα and CRABPII genes in Chinese cashmere goat skin during prenatal development

  • Zhong, Tao;Zhao, Wei;Zhou, Zhongqiang;Li, Li;Wang, Linjie;Li, Hua;Zhang, Hongping
    • Journal of Animal Science and Technology
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    • v.57 no.8
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    • pp.28.1-28.7
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    • 2015
  • Background: The physiologic characteristics of the cashmere trait and many of the differentially expressed genes relevant to hair cycling have been extensively studied, whereas genes involved in the prenatal development of hair follicles have been poorly investigated in cashmere goats. The aim of this study, therefore, was to quantify the time-course changes in the expressions of $TR{\alpha}$ and CRABPII genes in the fetal skin of Chinese cashmere goats at the multiple embryonic days (E70, E75, E80, E90, E100, E120 and E130) using real-time quantitative PCR (RT-qPCR). Results: RT-qPCR showed that $TR{\alpha}$ was expressed at E70 with relatively high level and then slightly decreased (E75, E80, and E90). The highest expression of $TR{\alpha}$ mRNA was revealed at E130 (P > 0.05). The expression pattern of CRABPII mRNA showed an 'up-down-up' trend, which revealed a significantly highest expression at E75 (P < 0.05) and was down-regulated during E80 to E120 (P < 0.05) and mildly increased at E130, subsequently. Conclusion: This study demonstrated that $TR{\alpha}$ and CRABPII genes expressed in different levels during prenatal development of cashmere. The present study will be helpful to provide the comprehensive understanding of $TR{\alpha}$ and CRABPII genes expressions during cashmere formation and lay the ground for further studies on their roles in regulation of cashmere growth in goats.

Anti-inflammatory Effects of the Fruits of Foeniculum vulgare in Lipopolysaccharide-stimulated Macrophages (대식세포에서 LPS로 유도된 염증에 대한 회향 열매의 항염 효과)

  • Yang, In Jun;Yu, Hak Yin;Lee, Dong-Ung;Shin, Heung Mook
    • Journal of Life Science
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    • v.24 no.9
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    • pp.981-987
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    • 2014
  • Foeniculum vulgare has long been prescribed in traditional medicine for the treatment of inflammation diseases. In this study, we aimed to investigate the inhibitory effects of the fruits of F. vulgare on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells under non-cytotoxic ($100{\mu}g/ml$) conditions. The 80% methanol extract was subsequently partitioned successively with hexane, methylene chloride, ethyl acetate, and n-butanol, and the fractions so obtained were also examined for their anti-inflammatory effects. Among them, the hexane, methylene chloride, and ethyl acetate fractions inhibited nitric oxide (NO) and prostaglandin E2 (PGE2) production in LPS stimulated macrophages. The methylene chloride and ethyl acetate fractions also suppressed the productions of interleukin $(IL)-1{\beta}$ and IL-6 by down-regulating their mRNA levels in LPS stimulated RAW 264.7 cells. Furthermore, the ethyl acetate fraction strongly suppressed tumor necrosis factor (TNF)-${\alpha}$ at the protein and mRNA levels in LPS stimulated RAW 264.7 cells. These observations suggest that the anti-inflammatory actions of F. vulgare are due to inhibitions of the productions of NO, PGE2, and pro-inflammatory cytokines.

EFFECTS OF SUBSTANCE P ON COLLAGEN PRODUCTION IN HUMAN PERIODONTAL LIGAMENT CELLS (치주인대 세포의 교원질 생성에 대한 Substance P의 효과)

  • CHUN, Jun-Yeung;Choi, Je-Yong;Kyung, Hee-Moon;Sung, Jae-Hyun
    • The korean journal of orthodontics
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    • v.26 no.1 s.54
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    • pp.83-94
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    • 1996
  • Substance P is one of the neuropeptide which presents highly in tension site of periodontal ligament during the orthodontic tooth movement. It has bnn also hon as one of the neuropeptides which cause neurogenic inflammation in various tissues and organs. However, there is no report about the effect of substance P on major extracellular matrix protein, collagen production. The purpose of this study was to evaluate the collagen production by substance P in human periodontal ligament cell. The collagenase-digestion method was used to evaluate collagen production and also used Northern blot hybridization for the evaluation of collagen mRNA level. This study also Included in terms of prostanglandins and gelatinase production with respect to collagen production. For the collagen degradation, zymography was used to estimate denatured collagen degradation. Dose-dependent effect of substance P on noncollagen protein, collagen, and percent collagen was that substance P increased noncollagen protein synthesis, but decreased collagen sytnsis. So the percent collagen, which determined by relative collagen production against total protein production, w3s decreased from $7\%\;to\;3.6\%$. This inhibitory effect of substance P on collagen production was disappeared when cells were treated concomitantly with indomethacin. It means that substance P-induced inhibitory effect on collagen production was due at least in part to the production of prostaglandins. To evaluate whether substance P-induced inhibitory effect on collagen production is correspond to the steady-state levels of procollagen mRNA, Northern blot hybridization was performed and it showed that substance P has no effect on the steady-slate level of ${\alpha}1(I)$ procollagen mRNA. It means that the inhibitory effect of substance P on collagen production was due to the change of a certain mechanism after posttranscription. In this context, gelatinase production by substance P in periodontal ligament cells was evaluated by zymography. Zymogram showed that substance P has no effect on gelatinase production in periodontal ligament cells. To explore wheter substance P-induced inhibitory effect on collagen production is selevtive in periodontal ligament cells or not, MC3T3-E1 cells which originated from mouse calvaria was used. It showed that substance P has no effect on collagen production in MC3T3-E1 cells. Taken together, substance P inhibits collagen production in human periodontal ligament cells. This effect was not due to the change of the steady-state level of procollagen mRNA and gelatinase production, but due at least in part to the change of prostaglandins production.

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Development of Probiotic Dairy Product for the Normalization of Microbial Flora in Korean Infants (한국인 영유아 장내균총 정상화를 위한 프로바이오틱 유제품의 개발)

  • Kim, Min-Kyung;Choi, A-Ri;Han, Gi-Sung;Jeong, Seok-Geun;Chae, Hyun-Seok;Jang, Ae-Ra;Seol, Kuk-Hwan;Oh, Mi-Hwa;Kim, Dong-Hun;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.290-295
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    • 2011
  • This study was conducted to develop an effective probiotic dairy product to normalize the microbial flora in Korean infants. A total of 2,200 colonies were isolated from 25 Korean neonates, and 16S rRNA of 348 isolates was analyzed. Approximately 40% of the lactic-acid producing bacterial isolates were Enterococcus faecalis, and 34.2% of them were strains similar to XR7 in the GenBank database. The fastest growing strain in MRS broth was registered as 91532 by the KACC. The selected strain was freeze-dried and utilized to ferment a milk-containing rice soup, tarakjuk. Microbiological, physico-chemical, and sensory characteristics of the fermented tarakjuk were compared with fermented milk and tarakjuk. E. faecalis KACC 91532 increased from $6.14{\pm}0.19$ to $7.36{\pm}0.13$ Log CFU/mL, and can be useful as a probiotic, as described in the Standards for Functional Health Foods.

Neuroprotection of Recombinant Human Erythropoietin Via Modulation of N-methyl-D-aspartate Receptors in Neonatal Rats with Hypoxic-ischemic Brain Injury (신생 백서의 저산소성 허혈성 뇌손상에서 NMDA receptor 조절을 통한 유전자 재조합 인 에리스로포이에틴의 신경보호)

  • Jang, Yoon-Jung;Seo, Eok-Su;Kim, Woo-Taek
    • Neonatal Medicine
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    • v.16 no.2
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    • pp.221-233
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    • 2009
  • Purpose: Erythropoietin (EPO) has neuroprotective effects in many animal models of brain injury, including hypoxic-ischemic (HI) encephalopathy, trauma, and excitotoxicity. Current studies have demonstrated the neuroprotective effects of EPO, but limited data are available for the neonatal periods. Here in we investigated whether recombinant human EPO (rHuEPO) can protect the developing rat brain from HI injury via modulation of NMDA receptors. Methods: In an in vitro model, embryonic cortical neuronal cell cultures from Sprague-Dawley (SD) rats at 19-days gestation were established. The cultured cells were divided into five groups: normoxia (N), hypoxia (H), and 1, 10, and 100 IU/mL rHuEPO-treated (H+E1, H+ E10, and H+E100) groups. To estimate cell viability and growth, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was done. In an in vivo model, left carotid artery ligation was performed on 7-day-old SD rat pups. The animals were divided into six groups; normoxia control (NC), normoxia Sham-operated (NS), hypoxia-ischemia only (H), hypoxia-ischemia+vehicle (HV), hypoxia-ischemia+rHuEPO before a HI injury (HE-B), and hypoxia-ischemia+rHuEPO after a HI injury (HE-A). The morphologic changes following brain injuries were noted using hematoxylin and eosin (H/E) staining. Real-time PCR using primers of subunits of NMDA receptors (NR1, NR2A, NR2B, NR2C and NR2D) mRNA were performed. Results: Cell viability in the H group was decreased to less than 60% of that in the N group. In the H+E1 and H+E10 groups, cell viability was increased to >80% of the N group, but cell viability in the H+E100 group did not recover. The percentage of the left hemisphere area compared the to the right hemisphere area were 98.9% in the NC group, 99.1% in the NS group, 57.1% in the H group, 57.0% in the HV group, 87.6% in the HE-B group, and 91.6% in the HE-A group. Real-time PCR analysis of the expressions of subunits of NMDA receptors mRNAs in the in vitro and in vivo neonatal HI brain injuries generally revealed that the expression in the H group was decreased compared to the N group and the expressions in the rHuEPO-treated groups was increased compared to the H group. Conclusion: rHuEPO has neuroprotective property in perinatal HI brain injury via modulation of N-methyl-D-aspartate receptors.

Role of Kupffer Cells in Vasoregulatory Gene Expression During Endotoxemia

  • Kim, Tae-Hoon;Lee, Sun-Mee
    • Biomolecules & Therapeutics
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    • v.16 no.4
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    • pp.306-311
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    • 2008
  • Although hepatic microcirculatory dysfunction occurs during endotoxemia, the mechanism responsible for this remains unclear. Since Kupffer cells provide signals that regulate hepatic response in inflammation, this study was designed to investigate the role of Kupffer cells in the imbalance in the expression of vasoactive mediators. Endotoxemia was induced by intraperitoneal E. coli endotoxin (LPS, 1 mg/kg body weight). Kupffer cells were inactivated with gadolinium chloride ($GdCl_3$, 7.5 mg/kg body weight, intravenously) 2 days prior to LPS exposure. Liver samples were taken 6 h following LPS exposure for RT-PCR analysis of mRNA for genes of interest: endothelin (ET-1), its receptors $ET_A$ and $ET_B$, inducible nitric oxide synthase (iNOS), heme oxygenase (HO-1), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$). mRNA levels for iNOS and TNF-$\alpha$ were significantly increased 31.8-fold and 26.7-fold in LPS-treated animals, respectively. This increase was markedly attenuated by $GdCl_3$, HO-1 expression significantly increased in LPS-treated animals, with no significant difference between saline and $GdCl_3$ groups. ET-1 was increased by LPS. mRNA levels for $ET_A$ receptor showed no change, whereas $ET_B$ transcripts increased in LPS-treated animals. The increase in $ET_B$ transcripts was potentiated by $GdCl_3$. We conclude that activation of Kupffer cells plays an important role in the imbalanced hepatic vasoregulatory gene expression induced by endotoxin.