• Title/Summary/Keyword: subsequent high dose

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Chronologic chrnge of serum IgG antibody response in chickens reinfected with Cryptosporidium baileyi (닭와포자충 재감염닭의 혈청1gG 항체가 추이)

  • Lee, Jae-Gu;Kim, Hyeon-Cheol;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.255-258
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    • 1996
  • Eight 2-day-old SPF Chickens were each inoculated Orally With 3 Sing1e dose Of 5 × 105 oocysts of Cryptosporinium boilevi. and immunoglobulin G (IgG) antibody responses were chronologically measured by indirect immunofluorescent antibody (IFA) assay. Anti-C. bcileyi IgG antibody levels remained high (1 : 106.67 to 1:512.00) for at least 4 months with 330 days of a detectable period. Ten days after the negative conversion, each chicken was re-challenged with 1 × 107 oocysts of the same species. Subsequent infection in 340-day-old individuals caused sudden elevated IgG antibody levels and the titer peaked on day 28 postchallenge inoculation (PCI), at 1:1.024 with a 65 days of detection period. Chickens in primary infection showed oocyst shedding profiles. but did not exhibit any oocyst shedding before or after experimental reinfection.

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Black soybean anthocyanins attenuate inflammatory responses by suppressing reactive oxygen species production and mitogen activated protein kinases signaling in lipopolysaccharide-stimulated macrophages

  • Kim, Jin Nam;Han, Sung Nim;Ha, Tae Joung;Kim, Hye-Kyeong
    • Nutrition Research and Practice
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    • v.11 no.5
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    • pp.357-364
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    • 2017
  • BACKGROUND/OBJECTIVES: Oxidative stress is closely related with inflammation and development of many diseases. Black soybean seed coat contains high amount of anthocyanins, which are well-known for free radical scavenging activities. This study investigated inflammatory response and action mechanism of black soybean anthocyanins with regard to antioxidant activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. MATERIALS/METHODS: RAW 264.7 cells were treated with anthocyanins extracted from black soybean seed coats in a concentration range of 12.5 to $100{\mu}g/mL$. The production of reactive oxygen species (ROS), secretion of pro-inflammatory mediators and cytokines, and the signaling in the mitogen activated protein kinases (MAPKs) pathway were examined. RESULTS: Black soybean anthocyanins significantly decreased LPS-stimulated production of ROS, inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$, and pro-inflammatory cytokines, including tumor necrosis factor ${\alpha}$ and interleukin-6, in a dose-dependent manner without cytotoxicity (P < 0.001). Black soybean anthocyanins downregulated the expression of inducible NO synthase and cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells (P < 0.001). Moreover, black soybean anthocyanins inhibited LPS-induced phosphorylation of MAPKs, including extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 (P < 0.001). CONCLUSION: These results suggest that black soybean anthocyanins exert anti-inflammatory activity by inhibiting ROS generation and subsequent MAPKs signaling, thereby inhibiting inflammatory responses.

Role of Intracellular Taurine in Monensin-induced $Na^+,\;Ca^{++}$ Accumulation and Mechanical Dysfunction in Isolated Rat Hearts

  • Kim, Young-Hoon;Park, Jong-Wan;Kim, Myung-Suk
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.5
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    • pp.537-546
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    • 1997
  • It has been postulated that the intracellular taurine is co-transported with $Na^+$down a concentration gradient and prevents the intracellular accumulation of sodium. It is therefore, expected that an elevated level of intracellular taurine prevents the sodium-promoted calcium influx to protect the cellular damages associated with sodium and calcium overload. In the present study, we evaluated the effects of intra- and extracellular taurine on the myocardial $Na^+$and$Ca^{++}$ contents and the cardiac functions in isolated rat hearts which were loaded with sodium by monensin, a $Na^+-ionophore$. Monensin caused a dose-dependent increase in intracellular $Na^+$ accompanied with a subsequent increase in intracellular $Ca^{++}$ and a mechanical dysfunction. In this monensin-treated heart, myocardial taurine content was decreased with a concomittent increase in the release of taurine. The monensin-induced increases in intracellular $Na^+$, $Ca^{++}$ and depression of cardiac function were prevented in the hearts of which taurine content had been increased by high-taurine diet. Conversely, in the hearts of which taurine concentration gradient had been decreased by addition of taurine in the perfusate, the monensin-induced increases in $Na^+$, $Ca^{++}$ and functional depression were accelerated. These results suggest that taurine, depending on the intra-extracellular concentration gradient, can affect intracellular sodium and calcium concentrations, and that an increased intracellular taurine may play a role in protection of myocardial dysfunction associated with the sodium and calcium overload.

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Differential Growth Response and Gene Expression in Relation to Capsidiol Biosynthesis of Red Pepper Plant and Cultured Cells by γ-Ray and UV Stress (방사선과 자외선에 대한 고추 식물체 및 배양세포의 생장반응과 Capsidiol 생합성 유전자의 발현 차이)

  • An, Jung-Hee;Kim, Jae-Sung;Jeong, Jeong-Hag;Oh, Sei-Myoung;Kwon, Soon-Tae
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.201-206
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    • 2003
  • Differential responses of red pepper plant and cultured cells to enhanced ${\gamma}$-ray($^{60}$ Co) and ultraviolet(UV) stress were investigated. In seed treatment, 1 Gy of ${\gamma}$-ray increased seedling dry weight up to 19.1%, but 50 Gy treatment markedly ingibited seed germination and subsequent growth of seedling. UV treatment to seed did not change the germination ability of seeds and the growth of seedlings regardless of duration of UV treatment until 24 hrs. In case of UV treatment to seedlings, plant injury was seriously progressed even after the seedlings were returned to no UV condition, and eventually all the leaves showed chlorosis by the stress. However, progress of plant injury by ${\gamma}$-ray stress slower than that caused by UV stress, and even at the high dose of ${\gamma}$-ray 50 Gy, did not caused the cholrosis of stressed plant leaf. Amount of electrolytes leakage from plant leaf by UV treatment for 24hrs was increased up to 28.8 folds in comparison with untreated control, whereas that of 50 Gy of ${\gamma}$-ray was increased only 1.2 folds. UV stress induced the production of capsidiol, antimicrobial phytoalexin, by activation of gene expression involved in capsidiol biosynthesis, such as sesquiterpene cyclase and cyclase and cytochrome P450 hydroxylase in the leaf and cultured cell, but ${\gamma}$-ray stress induced neither the production of capsidiol nor expression of the genes.

Human Neutrophil Elastase Inhibitory Alkaloids from Chelidonium majus L.

  • Kim, Jeong Yoon;Lee, Ji Hye;Song, Yeong Hun;Jeong, Won Min;Tan, Xuefei;Uddin, Zia;Park, Ki Hun
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.281-285
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    • 2015
  • Human neutrophil elastase (HNE) represents a good therapeutic target for the treatment of inflammatory diseases as well as invasion of microorganism. The methanol extract of a aerial part of Chelidonium majus L. showed high activity against the neutrophil elastase with an $IC_{50}$ value of $100{\mu}g/mL$. Due to its potency, subsequent bioactivity-guided fractionation of methanol extract led to six alkaloids (1-6), which were identified as dihydrosanguinarine (1), (s)-stylopine (2), arnottianamide (3), (+)-chelidonine (4), spallidamine (5), and N-trans-feruloyltyramine (6). Among of them, three alkaloids (2, 5, and 6) inhibited HNE in a dose-dependent manner with $IC_{50}$ ranging between 11.6 and $51.0{\mu}M$. Lineweaver-Burk and Dixon plots, and their secondary replots showed that alkaloids (2, 5, and 6) were mixed inhibitors of HNE. The analysis of $K_I$ and $K_{IS}$ value proved that all inhibitors (2, 5, and 6) had reversible mixed type I mechanism.

Apoptotic Effects of Sipimikwanjung-tang of Sasang Constitutional Medicine in Human Hepatoblastoma Cells

  • Song Seung-Yun;Bae Young-Chun;Lee Sang-Min;Kim Kyung-Yo;Joo Jong-Cheon;Ko Ki-Duk;Park Soo-Jeong;Lee Kyung-Sung;Choi Yong-Seok;Kim Jong-Yeol
    • The Journal of Korean Medicine
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    • v.26 no.1 s.61
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    • pp.46-58
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    • 2005
  • Objective : This study on Sipimikwanjung-tang was undertaken to evaluate its antioxidant capacities and antiperoxidation activities in rat liver tissues. Sipimikwanjung-tang which has been one of the prescriptions in sasang constitutional medicine is usually applied for the therapy of various liver diseases. It is elucidated that Sipimikwanjung-tang has antioxidants on liver tissue of rat and the cytotoxic effects on human hepatoblastoma Hep G2 cells. Methods: Sipimikwanjung-tang extract in antioxidant effects of Hep G2 cells is evaluated by MTT assay, DAPI staining, DNA fragmentation assays and FACS can analysis. Results: Sipimikwanjung-tang induced apoptosis in Hep G2 cells, and induced G1 and G2M arrest of the cell cycle as well as a significant increase in PARP and caspase-3 activity. It induced an increase in $H_2O_2$ generation and the subsequent $NF-{\kappa}B$ activation and also induced cell apoptosis through the caspase-3-dependent pathways in the low concentration of Sipimikwanjung-tang extracts. However, the high dose of Sipimikwanjung-tang extract in Hep G2 cells inhibited $TGF-{\beta}l-induced$ apoptosis via increase in cellular $H_2O_2$, formation and $NF-{\kappa}B$ activation in human hepatoblastoma Hep G2 cells. Conclusion: From this study, the possibility that Sipimikwanjung-tang extracts apply to antioxidant and apoptotic treatment of disease is revealed.

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Mucosal Immunization with Recombinant Adenovirus Encoding Soluble Globular Head of Hemagglutinin Protects Mice Against Lethal Influenza Virus Infection

  • Kim, Joo Young;Choi, Youngjoo;Nguyen, Huan H.;Song, Man Ki;Chang, Jun
    • IMMUNE NETWORK
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    • v.13 no.6
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    • pp.275-282
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    • 2013
  • Influenza virus is one of the major sources of respiratory tract infection. Due to antigenic drift in surface glycoproteins the virus causes annual epidemics with severe morbidity and mortality. Although hemagglutinin (HA) is one of the highly variable surface glycoproteins of the influenza virus, it remains the most attractive target for vaccine development against seasonal influenza infection because antibodies generated against HA provide virus neutralization and subsequent protection against the virus infection. Combination of recombinant adenovirus (rAd) vector-based vaccine and mucosal administration is a promising regimen for safe and effective vaccination against influenza. In this study, we constructed rAd encoding the globular head region of HA from A/Puerto Rico/8/34 virus as vaccine candidate. The rAd vaccine was engineered to express high level of the protein in secreted form. Intranasal or sublingual immunization of mice with the rAd-based vaccine candidates induced significant levels of sustained HA-specific mucosal IgA and IgG. When challenged with lethal dose of homologous virus, the vaccinated mice were completely protected from the infection. The results demonstrate that intranasal or sublingual vaccination with HA-encoding rAd elicits protective immunity against infection with homologous influenza virus. This finding underlines the potential of our recombinant adenovirus-based influenza vaccine candidate for both efficacy and rapid production.

Anti-inflammatory effect of Lycium barbarum on polarized human intestinal epithelial cells

  • Lee, So-Rok;Hwang, Hye-Jeong;Yoon, Ju-Gyeong;Bae, Eu-Young;Goo, Kyo-Suk;Cho, Sang-Joon;Cho, Jin Ah
    • Nutrition Research and Practice
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    • v.13 no.2
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    • pp.95-104
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    • 2019
  • BACKGROUND/OBJECTIVES: Inflammatory Bowel Disease (IBD) has rapidly escalated in Asia (including Korea) due to increasing westernized diet patterns subsequent to industrialization. Factors associated with endoplasmic reticulum (ER) stress are demonstrated to be one of the major causes of IBD. This study was conducted to investigate the effect of Lycium barbarum (L. barbarum) on ER stress. MATERIALS/METHODS: Mouse embryonic fibroblast (MEF) cell line and polarized Caco-2 human intestinal epithelial cells were treated with crude extract of the L. chinense fruit (LF). Paracellular permeability was measured to examine the effect of tight junction (TJ) integrity. The regulatory pathways of ER stress were evaluated in MEF knockout (KO) cell lines by qPCR for interleukin (IL) 6, IL8 and XBP1 spliced form (XBP1s). Immunoglobulin binding protein (BiP), XBP1s and CCAAT/enhancer-binding homologous protein (CHOP) expressions were measured by RT-PCR. Scanning Ion Conductance Microscopy (SICM) at high resolution was applied to observe morphological changes after treatments. RESULTS: Exposure to LF extract strengthened the TJ, both in the presence and absence of inflammation. In polarized Caco-2 pretreated with LF, induction in the expression of proinflammatory marker IL8 was not significant, whereas ER stress marker XBP1s expression was significantly increased. In wild type (wt) MEF cells, IL6, CHOP and XBP1 spliced form were dose-dependently induced when exposed to $12.5-50{\mu}g/mL$ extract. However, absence of XBP1 or $IRE1{\alpha}$ in MEF cells abolished this effect. CONCLUSION: Results of this study show that LF treatment enhances the barrier function and reduces inflammation and ER stress in an $IRE1{\alpha}$-XBP1-dependent manner. These results suggest the preventive effect of LF on healthy intestine, and the possibility of reducing the degree of inflammatory symptoms in IBD patients.

Strategic construction of mRNA vaccine derived from conserved and experimentally validated epitopes of avian influenza type A virus: a reverse vaccinology approach

  • Leana Rich Herrera-Ong
    • Clinical and Experimental Vaccine Research
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    • v.12 no.2
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    • pp.156-171
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    • 2023
  • Purpose: The development of vaccines that confer protection against multiple avian influenza A (AIA) virus strains is necessary to prevent the emergence of highly infectious strains that may result in more severe outbreaks. Thus, this study applied reverse vaccinology approach in strategically constructing messenger RNA (mRNA) vaccine construct against avian influenza A (mVAIA) to induce cross-protection while targeting diverse AIA virulence factors. Materials and Methods: Immunoinformatics tools and databases were utilized to identify conserved experimentally validated AIA epitopes. CD8+ epitopes were docked with dominant chicken major histocompatibility complexes (MHCs) to evaluate complex formation. Conserved epitopes were adjoined in the optimized mVAIA sequence for efficient expression in Gallus gallus. Signal sequence for targeted secretory expression was included. Physicochemical properties, antigenicity, toxicity, and potential cross-reactivity were assessed. The tertiary structure of its protein sequence was modeled and validated in silico to investigate the accessibility of adjoined B-cell epitope. Potential immune responses were also simulated in C-ImmSim. Results: Eighteen experimentally validated epitopes were found conserved (Shannon index <2.0) in the study. These include one B-cell (SLLTEVETPIRNEWGCR) and 17 CD8+ epitopes, adjoined in a single mRNA construct. The CD8+ epitopes docked favorably with MHC peptidebinding groove, which were further supported by the acceptable ∆Gbind (-28.45 to -40.59 kJ/mol) and Kd (<1.00) values. The incorporated Sec/SPI (secretory/signal peptidase I) cleavage site was also recognized with a high probability (0.964814). Adjoined B-cell epitope was found within the disordered and accessible regions of the vaccine. Immune simulation results projected cytokine production, lymphocyte activation, and memory cell generation after the 1st dose of mVAIA. Conclusion: Results suggest that mVAIA possesses stability, safety, and immunogenicity. In vitro and in vivo confirmation in subsequent studies are anticipated.

The Effects of Majarol on the Blood Pressure and Heart Rate in Rats and Isolated Frog Heart (Majarine 유도체인 majarol의 흰쥐에 있어서 혈압 및 심박동수에 대한 작용과 적출 개구리 심장에 대한 작용)

  • Lee Jong-Hwoa;Park Young-Hyun;Cho Byung-Heon;Kim Yu-Jae;Kim Jong-Bae;Kim Chun-Sook;Cha Young-Deog;Kim Young-Suk
    • The Korean Journal of Pharmacology
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    • v.22 no.1 s.38
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    • pp.34-44
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    • 1986
  • In our previous studies, we had clarified many pharmacological effects of majarine: the bacteriostatic effect in vitro; the potentiation of hypnotic action of alcohol; hypotensive effect in rats and hypothermic effect in mice. This study was undertaken to search for a new antihypertensive drug. Red crystalline was obtained from majarine (which was extracted from Berberis koreana Palibin) by chemical methods. And this crystalline was identified as $C_{19}H_{16}NO_4$ contained one hydroxy group instead of methoxy group of majarine in isoquinoline ring and named 'Majarol' (5,6-Dihydro-9-hydroxy, 10-methoxybenzo-[g]-1,3-benzodioxolo [5,6-a] quinolizinium). We examined the effects of majarol on blood pressure and heart rate in urethane ancsthetized rats and the rate and amplitude of contraction of isolated frog heart. Several drugs: atropine sulfate, diphenhydramine chloride, hexamethonium bromide, phentolamine, epinephrine, propranolol and isoproterenol were used to clarify the mechanism of the hypotensive action of majarol. The results of experimints were as follows; 1. In low dose (0.5-2mg/kg, i.v.), majarol showed a typical transient hypotensive effect and slight decrease in heart rate. In high dose (5-10 mg/kg, i.v.), majarol showed a typical transient and a subsequent prolonged hypotensive effect and a significant prolonged decrease in heart rate was followed. 2. The hypotensive effects of majarol was not abolished by the pretreatments with atropine sulfate, hexamethonium bromide and diphenhydramine. The pretreatment with phentolamine inhibited significantly the hypotensive effects of majarol and the pretreatment wtih majarol blocked markedly the hypertensive effect of epinephrine. The positive chronotropic effect of isoproterenol was not blocked by the pretreatment with majarol. 3. In low dose, majarol increased the amplitude and decreased rate of contraction, but in high dose, majarol inhibited the amplitude and rate of contraction of isolated frog heart.

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