• Title/Summary/Keyword: In vivo development

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Antioxidant and Anti-Cholesterol Activities of Standardized Ecklonia Stolonifera Extract (표준화된 곰피추출물의 항산화 활성 및 콜레스테롤 개선 효과)

  • Han, Xionggao;Kim, Woo-Hyeok;Choi, Sun-Il;Men, Xiao;Lee, Se-jeong;Jin, Heegu;Oh, Hyun-Ji;Kang, Dahye;Kim, HyungBin;Lee, Boo-Yong;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.36 no.4
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    • pp.353-362
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    • 2021
  • Ecklonia stolonifera, which belongs to the family Laminariaceae, is an edible perennial brown marine alga that is widely distributed, and is rich in polyphenols, including dieckol. Here, we investigated the radical scavenging activities of E. stolonifera extract (ESE) using various in vitro models. We further evaluated the effect of ESE on the cholesterol secretion inhibition activity in HepG2 cells, as well as the hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase activity. Our results showed that the total phenol, total flavonoid, and dieckol contents of ESE were 9.64±0.04 mg GAE/g, 2.72±0.08 mg RE/g and 27.42±0.66 mg/g, respectively. The antioxidant activity of ESE increased in a dose-dependent manner. In addition, the ESE inhibited cholesterol secretion from HepG2 cells with anti-HMG-CoA reductase activity. These results suggested that ESE possesses antioxidant and anti-cholesterol activities, and can therefore be used as a preclinical bioresource for development of health functional foods.

Gaseous signal molecule SO2 regulates autophagy through PI3K/AKT pathway inhibits cardiomyocyte apoptosis and improves myocardial fibrosis in rats with type II diabetes

  • Zhao, Junxiong;Wu, Qian;Yang, Ting;Nie, Liangui;Liu, Shengquan;Zhou, Jia;Chen, Jian;Jiang, Zhentao;Xiao, Ting;Yang, Jun;Chu, Chun
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.6
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    • pp.541-556
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    • 2022
  • Myocardial fibrosis is a key link in the occurrence and development of diabetic cardiomyopathy. Its etiology is complex, and the effect of drugs is not good. Cardiomyocyte apoptosis is an important cause of myocardial fibrosis. The purpose of this study was to investigate the effect of gaseous signal molecule sulfur dioxide (SO2) on diabetic myocardial fibrosis and its internal regulatory mechanism. Masson and TUNEL staining, Western-blot, transmission electron microscopy, RT-qPCR, immunofluorescence staining, and flow cytometry were used in the study, and the interstitial collagen deposition, autophagy, apoptosis, and changes in phosphatidylinositol 3-kinase (PI3K)/AKT pathways were evaluated from in vivo and in vitro experiments. The results showed that diabetic myocardial fibrosis was accompanied by cardiomyocyte apoptosis and down-regulation of endogenous SO2-producing enzyme aspartate aminotransferase (AAT)1/2. However, exogenous SO2 donors could up-regulate AAT1/2, reduce apoptosis of cardiomyocytes induced by diabetic rats or high glucose, inhibit phosphorylation of PI3K/AKT protein, up-regulate autophagy, and reduce interstitial collagen deposition. In conclusion, the results of this study suggest that the gaseous signal molecule SO2 can inhibit the PI3K/AKT pathway to promote cytoprotective autophagy and inhibit cardiomyocyte apoptosis to improve myocardial fibrosis in diabetic rats. The results of this study are expected to provide new targets and intervention strategies for the prevention and treatment of diabetic cardiomyopathy.

Efficient Anti-Tumor Immunotherapy Using Tumor Epitope-Coated Biodegradable Nanoparticles Combined With Polyinosinic-Polycytidylic Acid and an Anti-PD1 Monoclonal Antibody

  • Sang-Hyun Kim;Ji-Hyun Park;Sun-Jae Lee;Hee-Sung Lee;Jae-Kyung Jung;Young-Ran Lee;Hyun-Il Cho;Jeong-Ki Kim;Kyungjae Kim;Chan-Su Park;Chong-Kil Lee
    • IMMUNE NETWORK
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    • v.22 no.5
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    • pp.42.1-42.20
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    • 2022
  • Vaccination with tumor peptide epitopes associated with MHC class I molecules is an attractive approach directed at inducing tumor-specific CTLs. However, challenges remain in improving the therapeutic efficacy of peptide epitope vaccines, including the low immunogenicity of peptide epitopes and insufficient stimulation of innate immune components in vivo. To overcome this, we aimed to develop and test an innovative strategy that elicits potent CTL responses against tumor epitopes. The essential feature of this strategy is vaccination using tumor epitope-loaded nanoparticles (NPs) in combination with polyinosinic-polycytidylic acid (poly-IC) and anti-PD1 mAb. Carboxylated NPs were prepared using poly(lactic-co-glycolic acid) and poly(ethylene/maleic anhydride), covalently conjugated with anti-H-2Kb mAbs, and then attached to H-2Kb molecules isolated from the tumor mass (H-2b). Native peptides associated with the H-2Kb molecules of H-2Kb-attached NPs were exchanged with tumor peptide epitopes. Tumor peptide epitope-loaded NPs efficiently induced tumor-specific CTLs when used to immunize tumor-bearing mice as well as normal mice. This activity of the NPs significantly was increased when co-administered with poly-IC. Accordingly, the NPs exerted significant anti-tumor effects in mice implanted with EG7-OVA thymoma or B16-F10 melanoma, and the anti-tumor activity of the NPs was significantly increased when applied in combination with poly-IC. The most potent anti-tumor activity was observed when the NPs were co-administered with both poly-IC and anti-PD1 mAb. Immunization with tumor epitope-loaded NPs in combination with poly-IC and anti-PD1 mAb in tumor-bearing mice can be a powerful means to induce tumor-specific CTLs with therapeutic anti-tumor activity.

HPV 16 E6/E7 Transgenic Mice Have Hyperkeratosis and Modulated Antioxidant Enzyme Activities

  • Kim, Myoung-Ok;Lee, Eun-Ju;Kim, Sung-Hyun;Park, Jun-Hong;Cho, Kyoungin;Jung, Boo-Kyung;Kim, Hee-Chul;Hwnag, Sol-Ha;Kim, Sun-Jung;Ryoo, Zae-Young
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.106-106
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    • 2003
  • Human papillomavirus type 16(HPV16) has been known to the major factor for the development of uterine cervical carcinomas. We have extended these studies to investigate the in vivo activities of HPV-16 E6/E7 when expressed in squamous epithelia of transgenic mice. Grossly, hK14HPV16E6/E7 transgenic mice had multiple phenotypes, including wrinkled skin that was apparent prior to the appearance of hair on neonates, thickened ears, and loss of hair in adults. In the transgenic mice, the wrinkled skin phenotype on the body and legs died at the age of 3-4 weeks. Histological analysis of demonstrated that E6/E7 causes epidermal hyperplasia in multiple transgenic lineages with high penetrance. This epithelial hyperplasia was characterized by an expansion of the proliferating compartment and an expansion of the keratinocyte and was associated with hyperkeratosis. These transgenic mice expressed E6/E7 transgene mainly in skin, heart, pancreas and kidney. Hyperplasia was found at the skin. The enzyme activities of GR, GPx and CuZnSOD were measured from the transgene cause keratinocyte at the skin. The specific enzyme activities were significantly higher in transgenic mice skin compared to the normal mice skin. Thus these transgenic mice may be useful for the develpment of antioxidant enzymes or other therapies for HPV-associated hyperkeratosis.

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Soybean Sprout Rot Caused by Colletotrichum species (Colletotrichum species에 의한 콩나물 부패)

  • Kim, Yong-Ki;Ryu, Jae-Ki;Ryu, Jae-Dang;Lee, Sang-Yeop;Lee, Seong-Don
    • Research in Plant Disease
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    • v.8 no.3
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    • pp.175-178
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    • 2002
  • Two novel casual agents of soybean sprout rot occurred at soybean sprouts cultivated under structure in Suwon area in 1997 were isolated and their pathogenicity was tested in vivo. An isolate formed crowed, black acervuli which were oval to elongated with numerous black, needlelike, intermixed long and short setae, 65~110$\times$3.5~6.6 ${\mu}{\textrm}{m}$. Conidia were curved, lunate, unicellular and hyliane and measured 21.5~22.5$\times$3.5~4.0 ${\mu}{\textrm}{m}$. The other isolate produced conidia with straight and cylindrical, and measured 14.0~17.5$\times$3.5~4.5 ${\mu}{\textrm}{m}$. Apressorium size was measured 6.3~8.5$\times$4.5~5.0 ${\mu}{\textrm}{m}$. The agents were identified as Colletotrichum truncatum and C. gloeosporioides based on their morphological characteristics. There was a large difference in pathogenicity between two isolates. C. gloeosporioides caused dark brownish discoloration of whole plants. It showed high pathogenicity with severe disease development. Meanwhile C. gloeosporiodes caused light brown spots on cotyledon and its pathogenicity was not strong. The soybean sprout rot occurred by the two Colletotrichum species was firstly reported in soy-bean sprout in Korea, and we suggest it as “Colletotrichum rot of soybean sprout”.

Tetrahydrobiopterin Inhibits PDGF-stimulated Migration and Proliferation in Rat Aortic Smooth Muscle Cells via the Nitric Oxide Synthase-independent Pathway

  • Jiang, Xiaowen;Kim, Bo-Kyung;Lin, Haiyue;Lee, Chang-Kwon;Kim, Jung-Hwan;Kang, Hyun;Lee, Pil-Young;Jung, Seung-Hyo;Lee, Hwan-Myung;Won, Kyung-Jong
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.3
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    • pp.177-183
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    • 2010
  • Tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthase (NOS) activity, is known to play important roles in modulating both NO and superoxide production during vascular diseases such as atherosclerosis. However, the role of BH4 in functions of vascular smooth muscle cells is not fully known. In this study, we tested the effects of BH4 and dihydrobiopterin (BH2), a BH4 precursor, on migration and proliferation in response to platelet-derived growth factor-BB (PDGF-BB) in rat aortic smooth muscle cells (RASMCs). Cell migration and proliferation were measured using a Boyden chamber and a 5-bromo-2'-deoxyuridine incorporation assay, respectively, and these results were confirmed with an ex vivo aortic sprout assay. Cell viability was examined by 2,3-bis [2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assays. BH4 and BH2 decreased PDGF-BBinduced cell migration and proliferation in a dose-dependent manner. The inhibition of cell migration and proliferation by BH4 and BH2 was not affected by pretreatment with $N^G$-nitro-L-arginine methyl ester, a NOS inhibitor. Moreover, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by BH4 and BH2. Cell viability was not inhibited by BH4 and BH2 treatment. The present results suggest that BH4 and BH2 may inhibit PDGF-stimulated RASMC migration and proliferation via the NOS-independent pathway. Therefore, BH4 and its derivative could be useful for the development of a candidate molecule with an NO-independent anti-atherosclerotic function.

Development of Wound-treatment Formulation Using Plant Growth Regulators for Wound Healing of Some Tree Species (수피상처치료를 위한 식물생장조절물질 기반 상처유합제 개발)

  • Min, Hojung;Kim, Byeongkwan;Cha, Byeongjin
    • The Korean Journal of Pesticide Science
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    • v.20 no.2
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    • pp.83-92
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    • 2016
  • New tree wound-treatment formulations (WTF) were developed in this study. Stimulating effects of plant growth regulating substance on callus formation were evaluated in vivo twig disc culture of Zelkova serrata, Ziziphus jujuba, Pinus densiflora, Ginkgo biloba. and Aesculus turbinata. Based on the evaluation, WTF were prepared with 2,4-D, NAA, and IBA using xanthan gum as an extending agent. WTF were pasted on $2{\times}10cm$ artificial wound on the stem and the effects were evaluated 12 weeks later with the thickness of new callus-tissues. The effects varied with tree species and the growth regulators. In Z. serrata and A. turbinata, 2 mg/L of 2,4-D was the best WTF. In P. koraiensis and G. biloba, 8 mg/L of NAA and 1 mg/L of IBA were the best, respectively. Callus formation was quite lower in G. biloba compared to other species. Stimulating effect of thiophanate-methyl paste was excellent only in Z. serrata and lower than certain WTF in other species. Vaseline, which was used generally as an WTF, remained longer on the wound and causing decay. From the results, it could be recommended that 2,4-D, NAA and IBA were the best WTF for Z. serrata/A. turbinata, P. koraiensis and G. biloba, respectively.

Antibacterial Effects of Pseudomonas aeruginosa MB I-3 against Listonella anguillarum (어류 병원세균, Listonella anguillarum에 대한 Pseudomonas aeruginosa MB I-3의 항균 효과)

  • Lee, Su-Jung;Youn, I Na;Kim, Jin-Do;Lee, Jung Sick;Kim, Eunheui
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.17-24
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    • 2014
  • To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

The Immune Response of Mice Vaccinated with Japanese Encephalitis Vaccine, CJ50003 Produced in Vero Cells (베로 세포에서 생산된 2세대 일본뇌염 백신의 마우스에서의 면역원성)

  • ;;;;;;Kenneth H. Eckels
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.82-88
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    • 1999
  • In this study, to evaluate newly developed Japanese encephalitis (JE) vaccine candidate CJ50003, we assessed its immunogenicity along with a previously commercialized inactivated JE Biken vaccine. The CR0003 viral antigens produced in Vero cells were administered suhcutaneouly to mice either with alum-adjuvanled or free form. The ELISA titers and neutralizing (NEUV antibody titers accounting for major protective immunity in JE were determined. Mice given alum-adjuvanted vaccine had a 10 times higher antigen-specific NEUT antibody response than did those which {lad received free antigens. This NEUT antibody response was maintained until day 168 with NEUT titer more than 1:160. Even with the 0.5 ng of alum-adjuvanted antigen dose, NEUT titer was induced more than 1:10 which is considered as an evidence for seroconversion and protection. Thc mice immune sera had a similar rate of cross-reactivity against three different viral antigens, Nakayama-NlH, P3 and SA14; as determined by ELISA assay. In a mice challenge model, vaccination with the GI50003 conferred more protection than with commercialized Biken vaccine against Nakayama virus. These data demonstrated that CJ50003 vaccine candidate has an excellent prophylactic efficacy and implicated it has a strong potential for further development and commercialization.

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Normal Pregnancy of Mouse Embryos Transferred after Assisted Hatching by a 1.48$\mu\textrm{m}$ Diode Laser (1.48$\mu\textrm{m}$ Diode Laser로 보조 부화처리 후 이식된 생쥐배의 정상임신에 관한 연구)

  • 김은영;이봉경;남화경;이금실;윤산현;박세필;정길생;임진호
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.287-292
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    • 1998
  • The objective of this study was to test whether ZP drilling using a 1.48$\mu$m diode laser beam on mouse IVF embryos becomes effective the hatching and normal in vivo development, as a preliminary test for obtaining the additional proof that the 1.48$\mu$m diode laser could be used safely for human applications. The results obtained in this experiment were as follows: when the hatched rates of mouse embryos by laser ZP drilling according to the embryonic stage were examined until 72 hr (in case of blast tocyst: day 4 after IVF) or 120 hr (in case of 4-cell: day 2 after IVF) after treatment, the d data of laser drilled blastocysts (81.8%) was significantly higher than those of control (hatching blastocyst: day 4 after IVF) (54.2%) and laser drilled 4-cell embryos (45.5%) (p<0.05). When the effect of laser drilling on implantation rates following embryo transfer in day 3 synchronized pseudopregnant recipients was examined, the l laser drilled group (48.7%) was slightly higher than that of control group (43.6%). In addition, when the several pregnant mice delivered in two groups were analysed their chromosomal normality and tested reproductive ability, all p pups were presented normal chromosomal number (n=40) and showed normal growth and reproductive ability. Therefore, these results dem-onstrated that ZP drilling using a 1.48$\mu$m diode l laser can increase the embryo hatching and ind duce the normal pregnancy of mouse embryos.

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