• Title/Summary/Keyword: In Vitro Development

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Development of an Environmental Friend Pellet Coating Oil for Replacement of Antibiotics for Ruminant Aninmal (천연물질을 이용한 반추동물 항생제 대체용 친환경 펠렛코팅오일의 개발)

  • Choi, Bitna;Song, Wan-Sun;Choo, Byung Kil;Cho, Sangbuem;Ham, Young-Joo;Kim, Nam Hyung;Yang, Kyoung Hee;Kim, Young Jun;Lee, Hong-Gu;Choi, Nag-Jin
    • Korean Journal of Organic Agriculture
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    • v.23 no.2
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    • pp.373-390
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    • 2015
  • This study investigated the effect of coated feed using mixed oil on rumen fermentation characteristics. Two experiments were conducted based on materials that were mixed. First, cashew nut and soybean oils were mixed with white mineral oil. And second, different plant extracts were mixed with white mineral oil. At first experiment, inclusion levels of mixed oil on diet (0.03%, 0.1%, 0.5%) were applied as variables. A coated diet was fermented with rumen inoculum according to in vitro rumen fermentation and its parameters were investigated. In the result of first experiment, no negative effects on rumen pH were found. Significantly decreased dry matter digestibility was detected at 0.5% treatment (P<0.05). Total gas productions in control and 0.03% were significantly greater than those of others (P<0.05). Significantly reduced methane productions were found in all treatments compared to the control (P<0.05). Inclusion of mixed oil did not affect on ammonia production. Total volatile fatty acid production was also not influenced by coating with mixed oils. Rumen fermentation parameters were greatly changed according to introduced plant extracts at second experiment. The significantly lowest and greatest ammonia productions were found at treatments with Ixeris dentata and Plantago asiatica, respectively (P<0.05). The significantly greatest acetate and propionate productions were detected at treatments with Crucuma longa and Zizyphus vulgaris 1, respectively (P<0.05). All treatments, except Chrysanthemum idicum, Euyale ferox seed, Moringa leaf and fruit and Zizyphus vulgaris 1, showed significantly increased total volatile fatty acid production compared to the control (P<0.05). Only Paeonia lactiflora showed significantly lesser gas production than the control (P<0.05). In methane production, Ceramium, Zizyphus, Paeonia, Agrimonia, Torilis, Mugwort, Foeniculum, Euphorbia, Taraxacum, Artemisia, Momordica, Curcuma and Moringa reduced methane significantly compared to the control (P<0.05).

Occurrence of Bunch Rot Disease Caused by Aspergillus tubingensis on Shine Muscat Grape (Aspergillus tubingensis에 의한 샤인머스켓 포도송이썩음병(가칭)의 발생)

  • Kim, Young Soo;Kwon, Hyeok Tae;Hong, Seung-Beom;Jeon, Yongho
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.220-225
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    • 2019
  • During the year 2018, the symptoms of bunch rot on Shine Muscat (Vitis vinifera L.) were observed in Kimcheon-si, Gyeongbuk province in Korea. The disease appears on the Shine Muscat as a black rot due to prolific fungal sporulation after it has invaded into the Shine Muscat which look completely empty and dryness. Colonies of these fungi are present on the Shine Muscat skin from fruit setting and increase in amount from early season to harvest, while become peak at ripening stage. To isolate the causal agent, small fragments (2 to 3 mm) of decayed tissue from the lesion margin were placed onto potato dextrose agar (PDA) plates. Fungal colonies on PDA produced dense white aerial mycelium and then covered with dark black conidial heads. These heads were large and radiate, and vesicles were globose (2.12-32.0×2.0-3.1 ㎛). Based on morphological and cultural characteristics, this fungus was identified as Aspergillus tubingensis. To confirm its identity, the internal transcribed spacer, β-tubulin, and RNA polymerase II was sequenced for molecular identification. BLAST search indicated 99% identity with A. tubingensis. The pathogenicity test on healthy grape of Shine Muscat produced bunch rot, as the original symptoms. To select effective fungicides for the control of brunch rot, an in vitro antifungal activity of seven fungicides were evaluated against the growth of A. tubingensis. Five fungicides (dipenoconazole, tebuconazole, metconazole, iminoctadine, and captan) exhibited significantly strong suppression of the mycelial growth of A. tubingensis.

Involvement of Lysosome Membrane Permeabilization and Reactive Oxygen Species Production in the Necrosis Induced by Chlamydia muridarum Infection in L929 Cells

  • Chen, Lixiang;Wang, Cong;Li, Shun;Yu, Xin;Liu, Xue;Ren, Rongrong;Liu, Wenwen;Zhou, Xiaojing;Zhang, Xiaonan;Zhou, Xiaohui
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.790-798
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    • 2016
  • Chlamydiae, obligate intracellular bacteria, are associated with a variety of human diseases. The chlamydial life cycle undergoes a biphasic development: replicative reticulate bodies (RBs) phase and infectious elementary bodies (EBs) phase. At the end of the chlamydial intracellular life cycle, EBs have to be released to the surrounded cells. Therefore, the interactions between Chlamydiae and cell death pathways could greatly influence the outcomes of Chlamydia infection. However, the underlying molecular mechanisms remain elusive. Here, we investigated host cell death after Chlamydia infection in vitro, in L929 cells, and showed that Chlamydia infection induces cell necrosis, as detected by the propidium iodide (PI)-Annexin V double-staining flow-cytometric assay and Lactate dehydrogenase (LDH) release assay. The production of reactive oxygen species (ROS), an important factor in induction of necrosis, was increased after Chlamydia infection, and inhibition of ROS with specific pharmacological inhibitors, diphenylene iodonium (DPI) or butylated hydroxyanisole (BHA), led to significant suppression of necrosis. Interestingly, live-cell imaging revealed that Chlamydia infection induced lysosome membrane permeabilization (LMP). When an inhibitor upstream of LMP, CA-074-Me, was added to cells, the production of ROS was reduced with concomitant inhibition of necrosis. Taken together, our results indicate that Chlamydia infection elicits the production of ROS, which is dependent on LMP at least partially, followed by induction of host-cell necrosis. To our best knowledge, this is the first live-cell-imaging observation of LMP post Chlamydia infection and report on the link of LMP to ROS to necrosis during Chlamydia infection.

Comparison among the Sperm Preparation Methods on the Human Spermatozoa (인간정자에 있어서 정자처리법의 비교)

  • Pang, Myung-Geol;Chung, Ku-Min;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Chang, Yoon-Seok;Lee, Jin-Yong;Lee, Sang-Hoon;Chung, Yung-Chai;Kim, Chang-Keun
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.2
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    • pp.107-115
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    • 1993
  • Procedures to separate motile. normal & motile and acrosome-reacted sperm with high efficiency have clinical application in Assisted Reproductive Technology in terms of increasing the probability of fertilization by a normal sperm and subsequent normal embryonic development. This study evaluated the effects of 10 sperm preparation techniques [Swim-up from a washed pellet (SU). Swim-up from semen (SO). Continuous Percoll Gradients I (PIC). Discontinuous Percoll Gradients I (PID). Continuous Percoll Gradients II(P II C). Discontinuous Percoll Gradients II(P II D), SpermPrep (SFC). Wang's tube (WT). Albumin Gradients (AG), Low temperature capacitation (LTC)] on motility (%), normal morphology (%), motile sperm recovery rate(%). morphologically normal & motile sperm recovery rate (%), true acrosome reaction (%) and fertilizing ability. A P II D proved to be an effective means of separating morphologically normal & motile sperm. Our results indicated the P II D has advantages as compared with other methods in terms of recovery rate. enhancement of motility and normal morphology. And a LTC seems to be an effective means of enhancing the true acrosome reaction and fertilizing ability. These results suggest that the combined method of LTC and P II D for separation of morphologically normal & motile sperm and acrosome reacted sperm may be a useful procedure for intrauterine insemination and in vitro fertilization in the management of male factor infertility as well as for isolation of subpopulation of sperm for basic research.

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Study on Epidermal Growth Factor (EGF) and Expression of EGF-Receptor (EGF-R) in Mouse IVF/IVC Embryo;II. Expression of EGF-R on the Inner Cell Mass (ICM) of Mouse IVF/IVC Blastocyst (체외생산된 생쥐배에 대한 EGF와 EGF-R 발현에 관한 연구;II. 체외생산된 생쥐 배반포기배 ICM세포에서의 EGF-R 발현)

  • Kim, E.Y.;Kim, M.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.1
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    • pp.21-26
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    • 1997
  • This study was undertaken to examine the expression of EGF-R protein on ICM obtained from mouse IVF/IVC blastocyst by immunosurgery and indirect immunofluorescence(IIF). ICM cells used for this experiment were obtained from immunosurgery of mouse blatstocysts produced at 96 h after IVF, and recovered ICMs were assayed for cell viability and expression of EGF-R. The results obtained in this experiment were summarized as follows: when blastocysts were exposed to rabbit anti-mouse serum (antiserum) for 15-30 min. and then transferred them to guinea pig serum (complement) for 15-60 min., recovery rates of isolated ICMs were 8.0-84.2%. Especially, the best recovery (84.2%) of ICMs was obtained when exposure time to antiserum and complement was 30 min. and 60 min., respectively. In addition, when viability of isolated ICMs after immunosurgery was assessed by live/dead staining method, in all groups viability (93.8-100.0%) of isolated ICMs were not damaged if separated from TE cell. Also, we detected the expression of EGF-R on ICM cell by IIF. Therefore, these results suggest that EGF-R expression on the ICMs can stimulate the higher usablity of exogenous EGF to improve the preimplantation embryo development in vitro.

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Serum 8 Hydroxydeoxyguanosine and Cytotoxin Associated Gene A as Markers for Helicobacter pylori Infection

  • Yeniova, Abdullah Ozgur;Uzman, Metin;Kefeli, Ayse;Basyigit, Sebahat;Ata, Naim;Dal, Kursat;Guresci, Servet;Nazligul, Yasar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5199-5203
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    • 2015
  • Background: Helicobacter pylori (H.pylori) is associated with chronic gastritis, peptic ulcers, gastric adenocarcinomas and mucosa associated tissue lymphomas. Cytotoxin associated gene A (CagA) is one of the virulence factors of H.pylori. It is hypothesized that reactive oxygen species (ROS) play roles in H.pylori associated disease especially in development of gastric adenocarcinoma. Individuals infected with H.pylori bearing CagA produce more ROS than others. 8-hydroxydeoxyguanosine (8OHdG) is an in vitro marker of DNA damage and oxidative stress. The aim of this study was to investigate the relationship between 8OHdG level, H.pylori infection and CagA and alterations of serum 8OHdG level after H.pylori eradication. Materials and Methods: Patients admitted with dyspeptic complaints and upper gastrointestinal endoscopy were assessed. H.pylori was determined from histopathology of specimens. Serum 8OHdG levels of three groups (H.pylori negative, H. pylori positive CagA negative and H.pylori positive CagA positive) were compared. Patients with H.pylori infection received eradication therapy. Serum 8OHdG levels pretreatment and posttreatment were also compared. Results: In total, 129 patients (M/F, 57/72) were enrolled in the study. Serum 8OHdG level of H.pylori negative, H. pylori positive CagA negative and H.pylori positive CagA positive groups were significantly different ($5.77{\pm}1.35ng/ml$, $5.43{\pm}1.14ng/ml$ and $7.57{\pm}1.25ng/ml$ respectively, p=0.05). Furthermore, eradication therapy reduced serum 8OHdG level ($6.10{\pm}1.54ng/ml$ vs $5.55{\pm}1.23ng/ml$, p=0.05). Conclusions: Individuals infected with H.pylori bearing CagA strains have the highest serum 8OHdG level and eradication therapy decreases the serum 8OHdG level. To the best of our knowledge this is the first study that evaluated the effect of CagA virulence factor on serum 8OHdG level and the effect of eradication therapy on serum 8OHdG levels together. Eradication of CagA bearing H.pylori may prevent gastric adenocarcinoma by decreasing ROS. 8OHdG level may thus be a good marker for prevention from gastric adenocarcinoma.

Selection of Crabapple Pollinizers for 'Fuji' Apple through Physiological and Genetic Analysis (꽃사과 품종의 생리 및 유전적 분석을 통한 '후지' 사과의 수분수 선발)

  • Son, KwangMin;Choi, Dong Geun;Kwon, Soon-Il;Kim, Byung Oh;Choi, Cheol;Kang, In-Kyu
    • Journal of Bio-Environment Control
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    • v.22 no.2
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    • pp.116-122
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    • 2013
  • We investigated characteristics and self-incompatibility genotypes of 11 crabapple cultivars to introduce a new pollinizer of 'Fuji' apple tree in Korea. Flowering dates of eleven crabapples were two to seven days earlier than that of 'Fuji'. The rate of pollen germination in vitro was ranged from 85.6% to 98.0% except 'Virginia'. Controlled pollination treatment with each crabapples to 'Fuji' increased fruit set rate about 20.4% to 34.4%, the number of seed per fruit about 13.8% to 42.3% and fruit weight about 7.4% to 16.7% compared to open pollination. Tested crabapples were resistant to peach fruit moth, brown leaf spot and sooty blotch in general. A PCR amplification method using S-RNase primers was carry out in eleven crabapples. S-alleles, $S_3$, $S_5$, $S_9$, $S_{10}$, $S_{20}$, $S_{26} from six crabapples were determinated. Through sequencing analysis, $S_5$ ('Manchurian', 'Virginia') and $S_9$ ('Yantaishagou') showed 100% homologous to previous result. Based on our results, it was recommended that 'Manchurian', 'Hopa A', 'Hanyaehanakaidou', 'Spectabilis' could be promising pollinzers for 'Fuji' apple cultivar.

The Comparative Study of Immunomodulatory Effect by Glycyrrhiza New Varieties and Official Compendia (감초 신품종 및 약전 수재감초의 면역조절 효과 비교 연구)

  • Kang, Yun-Mi;Kim, Wonnam;Jin, Jong-Sik;Lee, Jong-Hyun;Chang, Jae Ki;Lee, Jeonghoon;An, Hyo-Jin
    • The Korea Journal of Herbology
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    • v.35 no.6
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    • pp.11-19
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    • 2020
  • Objective : The genus Glycyrrhiza has been used in food and traditional herbal medicine. Glycyrrhiza new varieties Wongam and Sinwongam have been developed by Korea Rural Development Administration and investigated to register on Korean Pharmacopoeia of the Ministry of Food and Drug Safety. The aim of this study is to investigate the immunomodulatory effect of Wongam and Sinwongam comparing with listed Glycyrrhiza species (Glycyrrhiza uralensis Fischer and G. glabra Linne) for evaluations about pharmacological effect of Glycyrrhiza new varieties. Methods : We studied the immunomodulatory effect of Wongam and Sinwongam compared with G. uralensis and G. glabra using THP-1 cell in vitro model. The cells were treated with phorbol 12-myristate 13-acetate (PMA) for differentiation and stimulated with lipopolysaccharides (LPS) to induce immune activation. We analyzed and compared the effects Glycyrrhiza new varieties and listed Glycyrrhiza species using nitric oxide (NO) assay, western blot, and reverse transcription-quantitative polymerase chain reaction analysis. 1) Results : Wongam and Sinwongam showed no cytotoxicity in THP-1 cells. Wongam and Sinwongam, and listed Glycyrrhiza species increased NO production, and cyclooxygenase (COX)-2 expression with or without LPS in differentiated THP-1 macrophages. Furthermore, Wongam and Sinwongam and listed Glycyrrhiza species upregulated the mRNA expressions of T helper type 1 (Th 1)-associated cytokines in LPS-stimulated THP-1 macrophages. Conclusion : These results indicated that Wongam and Sinwongam would have effect of enhancing immune response through the increase of NO and COX-2 expression, and activate Th1-associated cytokines. The findings of this study suggest the wide applicability of Glycyrrhiza new varieties.

Ginsenoside Re Inhibits Osteoclast Differentiation in Mouse Bone Marrow-Derived Macrophages and Zebrafish Scale Model

  • Park, Chan-Mi;Kim, Hye-Min;Kim, Dong Hyun;Han, Ho-Jin;Noh, Haneul;Jang, Jae-Hyuk;Park, Soo-Hyun;Chae, Han-Jung;Chae, Soo-Wan;Ryu, Eun Kyoung;Lee, Sangku;Liu, Kangdong;Liu, Haidan;Ahn, Jong-Seog;Kim, Young Ock;Kim, Bo-Yeon;Soung, Nak-Kyun
    • Molecules and Cells
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    • v.39 no.12
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    • pp.855-861
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    • 2016
  • Ginsenosides, which are the active materials of ginseng, have biological functions that include anti-osteoporotic effects. Aqueous ginseng extract inhibits osteoclast differentiation induced by receptor activator of NF-${\kappa}B$ ligand (RANKL). Aqueous ginseng extract produces chromatography peaks characteristic of ginsenosides. Among these peaks, ginsenoside Re is a major component. However, the preventive effects of ginsenoside Re against osteoclast differentiation are not known. We studied the effect of ginsenoside Re on osteoclast differentiation, RANKL-induced tartrate-resistant acid phosphatase (TRAP) activity, and formation of multinucleated osteoclasts in vitro. Ginsenoside Re hampered osteoclast differentiation in a dose-dependent manner. In an in vivo zebrafish model, aqueous ginseng extract and ginsenoside Re had anti-osteoclastogenesis effects. These findings suggest that both aqueous ginseng extract and ginsenoside Re prevent bone resorption by inhibiting osteoclast differentiation. Ginsenoside Re could be important for promoting bone health.

Effects of Medium Compositions and Plant Growth Regulators on in vitro Organogenesis in Cultured Explants of Platycodon grandiflorum Species

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Kim, Hye-Rim;Moon, Young-Ja;Yoon, Ki-Hong;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.3
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    • pp.259-274
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    • 2017
  • Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on 1/4 MS for P. grandiflorum with wild and green petals and on 1/8 MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on 1/4 MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, 1/4 MS supplemented with $1mg\;L^{-1}$ 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on $0.5mg{\cdot}L^{-1}$ thidiazuron (TDZ) from double petal explants grown on 1/8 MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.