• 한국수의병리학회지
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• 제2권1호
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• pp.1-8
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• 1998

Chickens at 3-weeks of age were inoculated with a highly virulent strain (SH/92) of Infectious Bursal Disease Virus(IBDV) through ocular and cloacal routes. The infected chickens were killed at 6, 12, 24, 48, 72, and 96 hrs post inoculation (PI) and Bursa of Fabricius(BF) were collected. The sizes of bursal follicules in infected chickens decreased at 48 to 96 hrs PI. Histologically the cellular changes were first evident at 12 hrs PI and characterized by condensation of nuclear chromatin of bursal lymphocytes indicating apoptosis. By 24 hrs PI apoptotic lymphocytes dramatically increased. In addition infiltration of heterophils were also seen in the follicles and in the interfollicular connective tissues. At 48 hrs PI, cystic cavities were observed in the follicles. As the infection advanced the bursal follicles showed atrophy accompanied by disappearance of heterophils and reduction in number of lymphocytes in the cystic cavities which was replaced by proteineous materials. The nuclei of most affected lymphocyte stained positively with the in situ end labeling for apoptosis. Electron microscopy showed viral particles with crystalline array in the lymphocytes of BF infected with IBOV. These results indicated that SH/92 IBDV infection in chickens caused increased apoptosis in the BF.

• Environmental Analysis Health and Toxicology
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• 제21권2호
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• pp.127-138
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• 2006

In urban areas, diesel exhaust particles (DEP) are probably a major component of particulate matters, especially in Korea where drive many diesel vehicles. The aim of this study was to investigate genotoxic effects of DEP using single ceil gel electrophoresis. In order to evaluate the mechanisms of DEP genotoxicity, the rat microsome mediated and DNA repair enzyme treated comet assays together with conventional comet assay were performed. Total diesel particles (DEPT) was collected without site fractionation from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEPT revealed DNA damage itself in NIH/3T3 cells. The level of DNA breaks plus oxidative DNA lesions and microsome mediated DNA damage was assessed by modified single cell gel eletrophoresis. DEPT was able to induce oxidative DNA damage as well as microsome mediated DNA damage. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor. reduced DNA damage in the presence of S-9 mixture. $DEP_T$ is the sources of oxidative stress, but antioxidants can significantly reduce oxidative DNA dmage. And $DEP_T$ may contain indirect mutagens which can be inhibited by CYP1A1 inhibitors. The ethanol extracts of the mixed vegetables (BV) or the mixed fruits (BF) were evaluated for their in vitro antigenotoxic effects. BV and BF showed potent Inhibitory effects against DEPT induced DNA damage with oxidative DNA lesions and in the prescence of S-9 mixture. These results indicate that BV and BF could prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P4501A1 in cell culture.

• Biomolecules & Therapeutics
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• 제23권4호
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• pp.367-373
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• 2015

Cordyceps species including Cordyceps bassiana are a notable anti-cancer dietary supplement. Previously, we identified several compounds with anti-cancer activity from the butanol fraction (Cb-BF) of Cordyceps bassiana. To expand the structural value of Cb-BF-derived anti-cancer drugs, we employed various chemical moieties to produce a novel Cb-BF-derived chemical derivative, KTH-13-amine-monophenyl [4-isopropyl-2-(1-phenylethyl) aniline (KTH-13-AMP)], which we tested for anti-cancer activity. KTH-13-AMP suppressed the proliferation of MDA-MB-231, HeLa, and C6 glioma cells. KTH-13-AMP also dose-dependently induced morphological changes in C6 glioma cells and time-dependently increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, the levels of the active full-length forms of caspase-3 and caspase-9 were increased. In contrast, the levels of total forms of caspases-3, caspase-8, caspase-9, and Bcl-2 were decreased in KTH-13-AMP treated-cells. We also confirmed that the phosphorylation of STAT3, Src, and PI3K/p85, which is linked to cell survival, was diminished by treatment with KTH-13-AMP. Therefore, these results strongly suggest that this compound can be used to guide the development of an anti-cancer drug or serve as a lead compound in forming another strong anti-proliferative agent.

• The Korean Journal of Physiology and Pharmacology
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• 제20권3호
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• pp.253-259
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• 2016

Previously, we found that KTH-13 isolated from the butanol fraction of Cordyceps bassiana (Cb-BF) displayed anti-cancer activity. To improve its antiproliferative activity and production yield, we employed a total synthetic approach and derivatized KTH-13 to obtain chemical analogs. In this study, one KTH-13 derivative, 4-(tert-butyl)-2,6-bis(1-phenylethyl)phenol (KTH-13-t-Bu), was selected to test its anti-cancer activity. KTH-13-t-Bu diminished the proliferation of C6 glioma, MDA-MB-231, LoVo, and HCT-15 cells. KTH-13-t-Bu induced morphological changes in C6 glioma cells in a dose-dependent manner. KTH-13-t-Bu also increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, KTH-13-t-Bu increased the levels of cleaved caspase-3 and -9. In contrast, KTH-13-t-Bu upregulated the levels of pro- and cleaved forms of caspase-3, -8, and -9 and Bcl- 2. Phospho-STAT3, phospho-Src, and phospho-AKT levels were also diminished by KTH13-t-Bu treatment. Therefore, these results strongly suggest that KTH-13-t-Bu can be considered a novel anti-cancer drug displaying pro-apoptotic activity.

• Animal Bioscience
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• 제37권8호
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• pp.1440-1451
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• 2024

Objective: This study aimed to develop and evaluate the effectiveness of a water-soluble microencapsulation method for probiotic strains using gum Arabic (GA) and skim milk (SKM) over a three-month storage period following processing. Methods: Four strains of Pediococcus acidilactici (BYF26, BYF20, BF9, and BF14) that were typical lactic acid bacteria (LAB) isolated from the chicken gut were mixed with different ratios of GA and SKM as coating agents before spray drying at an inlet temperature 140℃. After processing, the survivability and probiotic qualities of the strains were assessed from two weeks to three months of storage at varied temperatures, and de-encapsulation was performed to confirm the soluble properties. Finally, the antibacterial activity of the probiotics was assessed under simulated gastrointestinal conditions. Results: As shown by scanning electron microscopy, spray-drying produced a spherical, white-yellow powder. The encapsulation efficacy (percent) was greatest for a coating containing a combination of 30% gum Arabic: 30% skim milk (w/v) (GA:SKM30) compared to lower concentrations of the two ingredients (p<0.05). Coating with GA:SKM30 (w/v) significantly enhanced (p<0.05) BYF26 survival under simulated gastrointestinal conditions (pH 2.5 to 3) and maintained higher survival rates compared to non-encapsulated cells under an artificial intestinal juices condition of pH 6. De-encapsulation tests indicated that the encapsulated powder dissolved in water while keeping viable cell counts within the effective range of 10⁶ for 6 hours. In addition, following three months storage at 4℃, microencapsulation of BYF26 in GA:SKM30 maintained both the number of viable cells (p<0.05) and the preparation's antibacterial efficacy against pathogenic bacteria, specifically strains of Salmonella. Conclusion: Our prototype water-soluble probiotic microencapsulation GA:SKM30 effectively maintains LAB characteristics and survival rates, demonstrating its potential for use in preserving probiotic strains that can be used in chickens and potentially in other livestock.

• 한국식품영양과학회지
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• 제44권3호
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• pp.356-362
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• 2015

제주산 손바닥 선인장의 엽상경을 열수와 70% 에탄올로 추출한 후 분획하여 총 페놀 및 총 플라보노이드 함량과 항산화 활성 및 항염증 활성을 측정하였다. 총 페놀 함량은 에틸 아세테이트 분획물이 784 mg GAE/g으로 가장 높았고 그 다음으로 부탄올 분획물(452), 헥산 분획물(220) 순이었다. 총 플라보노이드 함량도 에틸아세테이트 분획물이 214 mg GE/g으로 가장 높았고 헥산 분획물(113), 부탄올 분획물(76) 순이었다. 에틸아세테이트 분획물, 부탄올 분획물, 헥산 분획물의 DPPH 라디칼 소거능($IC_{50}$)은 각각 103, 359, $469{\mu}g/mL$였고, ABTS 라디칼 소거능($IC_{50}$)도 각각 105, 379, $605{\mu}g/mL$로 에틸아세테이트 분획물이 가장 높았다. 유해산소 억제 능력(ORAC)은 에틸아세테이트 분획물이 $391{\mu}M$ TE로 가장 높은 활성을 보였고 부탄올 분획물(117), 헥산 분획물(64) 순이었지만, superoxide anion 소거 활성($IC_{50}$, ${\mu}g/mL$)은 에틸아세테이트 분획물(40), 부탄올 분획물(69), 70% 에탄올(98) 순이었다. RAW264.7 세포에 의한 NO 생성 저해능($IC_{50}$, ${\mu}g/mL$)은 헥산 분획물(62), 에틸아세테이트 분획물(104), 부탄올 분획물(465) 순으로 높은 활성을 보였다. 세포독성 대비 NO 생성 저해 활성을 나타내는 SI 지수는 에틸아세테이트 분획물이 4.63으로 가장 높았고, 다음으로는 헥산 분획물(3.37), 부탄올 분획물(2.14), 증류수 추출물(1.66) 순으로, 양성 대조군으로 사용한 quercetin(6.25)보다는 낮았다. 결론적으로 제주산 손바닥 선인장 엽상경의 추출물과 분획물 중에서 에틸아세테이트 분획물이 항산화 활성과 항염 활성이 가장 높았으며, 이는 총 페놀과 총 플라보노이드 함량에 기인하는 것으로 추정되었다.

• Journal of Microbiology and Biotechnology
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• 제19권3호
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• pp.331-337
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• 2009

Interleukin-13 (IL-13) has been proposed as a therapeutic target for bronchial asthma as it plays crucial roles in the pathogenesis of the disease. We developed an in vitro test system measuring transcriptional downregulatory activities on IL-13 as a primary screening method to select drug candidates from natural products. The promoter region of IL-13 (-2,048 to +1) was cloned into the upstream of a luciferase gene in the plasmid pGL4.14 containing the hygromycin resistance gene as a selection marker, generating pGL4.14-IL-13. The EL-4 thymoma and RBL-2H3 mast cells transiently expressing this plasmid highly produced the luciferase activities by responding to PI (PMA and ionomycin) stimulation up to 8-fold and 13-fold compared with the control, respectively, whereas cyclosporin A, a well-known antiasthmatic agent, significantly downregulated the activities. The BF1 clone of RBL-2H3 cells constitutively expressing pGL4.14-IL-13 was established by selecting surviving cells under a constant lethal dose of hygromycin treatment. The feasibility of this system was evaluated by measuring the downregulatory activities of 354 natural products on the IL-13 promoter using the BF1 clone. An extract from Morus bombycis (named TBRC 156) significantly inhibited PI-induced luciferase activities and IL-13 mRNA expression, but not the protein expression. Fisetin (named TBRC 353) inhibited not only PI-induced luciferase activities and mRNA expression, but also the IL-13 protein secretion, whereas myricetin (named TBRC 354) could not suppress the IL-13 expression at all. Our data indicated that this in vitro test system is able to discriminate the effects on IL-13 expression, and furthermore, that it might be suitable as a simple and time-saving primary screening system to select antiasthmatic agents by measuring transcriptional activities of the IL-13 promoter.

• 한국어병학회지
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• 제33권1호
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• pp.55-62
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• 2020

본 연구에서는 점액포자충에 의한 넙치의 여윔증 치료 후보물질을 탐색하기 위하여, Coxi-stop, Coxiclin 및 BLEAN80 등의 제품을 사용하여 in vitro 및 in vivo 실험을 실시하였다. 톨트라주릴이 주성분인 Coxi-stop의 경우, BF-2 cell을 사용한 in vitro 실험에서 점액포자충의 활성을 감소시키는 결과를 보였고, cell lysis와 같은 세포 독성 현상이 나타나지 않았다. In vivo 실험에서 Coxi-stop을 사용하여 약욕한 실험군은 대조군에 비하여 누적폐사율이 낮게 나타났으며, 이것은 톨트라주릴이 어류 기생충성 질병에 치료 효과가 있다는 기존의 보고와 유사한 결과이다. 본 연구에서는 톨트라주릴이 넙치의 여윔증 치료제로서 가능성이 있는 후보 물질이라는 것을 제시한다.

• 한국어병학회지
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• 제30권2호
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• pp.71-78
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• 2017

The glycoprotein of novirhabdoviruses is known to play a critical role in the determination of host specificity. Viral hemorrhagic septicemia viruses (VHSVs) in different genotypes have different glycoprotein sequences and show different preferences for specific cell lines. In this study, to know whether the glycoprotein is solely responsible for the host cell preference of VHSV, a recombinant VHSV expressing vesicular stomatitis virus (VSV) glycoprotein instead of VHSV IVa glycoprotein (rVHSV-VSV-G) was generated by reverse genetics and inoculated into several fish cell lines, then, cytopathic effect (CPE) and viral growth caused by rVHSV-VSV-G infection were compared with those caused by rVHSV-wild that was previously generated and has the same genomic sequence with wild-type VHSV except a few nucleotides. The plaque numbers of rVHSV-VSV-G were significantly higher in EPC, BF-2 and GF cells than those of rVHSV-wild. However, in HINAE cells (originated from olive flounder), rVHSV-VSV-G titer was significantly lower than rVHSV-wild titer, and both recombinant VHSVs were not grown well in CHSE-214 cells. Although statistical significances were detected in the titers between rVHSV-wild and rVHSV-VSV-G in several cell lines, the cell line-preference order of rVHSV-VSV-G was not different from that of rVHSV-wild. These results suggest that the replacement of VHSV glycoprotein may not completely change host cell preference, and other regions of VHSV might also involve in the determination of host cell preference.

• 한국어병학회지
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• 제23권3호
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• pp.335-342
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• 2010

2008년 3월 인도네시아로부터 수입된 paradise fish에서 대량폐사가 발생하여 병원체 검사를 실시한 결과, megalocytivirus 및 Mycobacterium sp.가 검출되었으며, CHSE-214세포에서 미동정 바이러스가 분리되었다. 본 연구에서는 미동정 바이러스의 특성을 조사하였으며, 또한 병원성 실험을 통해 paradise fish의 대량 폐사와의 연관성을 조사하였다. 분리 바이러스는 CHSE-214, BF-2, GF, SSN-1, FSP 및 FFN 세포에서 합포체의 세포변성효과를 나타내었고 IUdR 및 chloroform 처리에 감염성이 상실되지 않으며, 산 (pH 3), 알카리 (pH 11) 및 열 ($56^{\circ}C$, 30분)에 안정하였다. 핵산 분석 결과에서는 적어도 10개의 segment (0.7-3.6 kb)를 지닌 RNA 바이러스로 확인되었으며, 전자현미경 관찰 결과, 약 65 nm 크기로 인벨롭이 없으며 2중의 capsid를 지닌 바이러스가 세포질에서 관찰되었다. 이상의 결과는 Reoviridae family에 속하는 바이러스의 전형적인 특성과 일치하므로 paradise fish에서 분리된 바이러스는 reovirus로 확인되었다. Reovirus를 사용한 감염실험에서는 병원성이 나타나지 않아, reovirus는 paradise fish의 대량 폐사의 직접적인 원인이 아닐 것으로 사료되었다.