• Title/Summary/Keyword: In vitro inoculation

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Synthesis and Antitumor Activity of 3'-hydroxydaunorubicin and 3'-hydroxydoxorubicin Derivatives Substituted with 4'-fluorine or 4'-azide in Sugar Moiety (당부분에서 4'-플루오린 또는 4'-아자이드로 치환된 3'-히드록시다우노루비신과 3'-히드록시독소루비신 유도체의 합성과 항암활성)

  • Ok, Kwang-Dae;Park, Jeong-Bae;Kim, Moon-Sung;Jung, Dong-Yoon;Yang, Junn-Ick
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.117-125
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    • 1996
  • 3'-Deamino-3-hydroxy-4'-fluoro- or 3'-deamino-3'-hydroxy-4'-azido-daunorubicin(6,8) and -doxorubicin(7,9) have been synthesized, respectively. Compounds 7,8 and 9 were mo re cytotoxic than daunorubicin(1) and doxorubicin(2) against L1210 murine leukemic cell in vitro. When administered intraperitoneally for 9 days starting 1 day after tumor inoculation, compounds 7(T/C 605%) and 9(T/C 488%) showed significant antitumor activity for ip-inoculuated L1210 murine leukemia at wide range of doses.

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Development of Cryptosporidium parvum in cell culture (세포배양에서 Cryptosporidium parvum의 발육)

  • Kim, Bo-sook;Joo, Hoo-don;Wee, Sung-hwan;Kim, Tae-jong
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.317-326
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    • 1995
  • The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

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Resistance Induction by Salicylic Acid Formulation in Cassava Plant against Fusarium solani

  • Saengchan, Chanon;Phansak, Piyaporn;Thumanu, Kanjana;Siriwong, Supatcharee;Le Thanh, Toan;Sangpueak, Rungthip;Thepbandit, Wannaporn;Papathoti, Narendra Kumar;Buensanteai, Natthiya
    • The Plant Pathology Journal
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    • v.38 no.3
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    • pp.212-219
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    • 2022
  • Fusarium root rot caused by the soil-borne fungus Fusarium solani is one of the most important fungal diseases of cassava in Thailand, resulting in high yield losses of more than 80%. This study aimed to investigate if the exogenous application of salicylic acid formulations (Zacha) can induce resistance in cassava against Fusarium root rot and observe the biochemical changes in induced cassava leaf tissues through synchrotron radiation based on Fourier-transform infrared (SR-FTIR) microspectroscopy. We demonstrated that the application of Zacha11 prototype formulations could induce resistance against Fusarium root rot in cassava. The in vitro experimental results showed that Zacha11 prototype formulations inhibited the growth of F. solani at approximately 34.83%. Furthermore, a significant reduction in the disease severity of Fusarium root rot disease at 60 days after challenge inoculation was observed in cassava plants treated with Zacha11 at a concentration of 500 ppm (9.0%). Population densities of F. solani were determined at 7 days after inoculation. Treatment of the Zacha11 at a concentration of 500 ppm resulted in reduced populations compared with the distilled water control and differences among treatment means at each assay date. Moreover, the SR-FTIR spectral changes of Zacha11-treated epidermal tissues of leaves had higher integral areas of lipids, lignins, and pectins (1,770-1,700/cm), amide I (1,700-1,600/cm), amide II (1,600-1,500/cm), hemicellulose, lignin (1,300-1,200/cm), and cellulose (1,155/cm). Therefore, alteration in defensive carbohydrates, lipids, and proteins contributed to generate barriers against Fusarium invasion in cassava roots, leading to lower the root rot disease severity.

Enhancement of Nutritional Quality of Italian Ryegrass Mediated Silage by Supplemented with Lactic Acid Bacteria and Chlorella

  • Vijayakumar, Mayakrishnan;Ilavenil, Soundarrajan;Arasu, Mariadhas Valan;Jung, Min-Woong;Park, Hyung Soo;Kim, Ji Hea;Lim, Young Cheol;Choi, Ki Choon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.3
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    • pp.169-173
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    • 2014
  • The aim of present study was to improve the quality of silage using lactic acid bacteria (LAB) and chlorella as a supplement. Italian ryegrass (IRG) mediated silage was prepared with lactic acid bacteria (L. plantarum) and different concentration of chlorella. We analyzed the nutritional profiles such as crude protein (CP), acid detergent fiber (ADF) neutral detergent fiber (NDF), total digestible nutrient (TDN) and in-vitro dry matter digestibility (IVDMD), microbial counts and fermentative acids such as lactic acid, acetic acid and butyric acid in the control and experimental silage after three months. It shows increased crude protein content and also maintains the rest of nutritional values as compared with control silage. LAB inoculation with chlorella as supplementation slightly reduced the pH of the silage. In addition, it increased the fermentative acids production as compared with control silage and inhibits the undesired microbial growth especially fungi in the silage. Therefore, we suggest that LAB inoculation and chlorella supplementation to the IRG mediated silage could be improved the nutritional quality of the silage which is an intrinsic feature for the application in the preparation of animal feeds and functional foods.

Changes in fermentation pattern and quality of Italian ryegrass (Lolium multiflorum Lam.) silage by wilting and inoculant treatments

  • Liu, Chang;Zhao, Guo Qiang;Wei, Sheng Nan;Kim, Hak Jin;Li, Yan Fen;Kim, Jong Geun
    • Animal Bioscience
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    • v.34 no.1
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    • pp.48-55
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    • 2021
  • Objective: This study was conducted to investigate the effects of wilting and microbial inoculant treatment on the fermentation pattern and quality of Italian ryegrass silage. Methods: Italian ryegrass was harvested at heading stage and ensiled into vinyl bags (20 cm×30 cm) for 60d. Italian ryegrass was ensiled with 4 treatments (NWNA, no-wilting noadditive; NWA, no-wilting with additive; WNA, wilting no-additive; WA, wilting with additive) in 3 replications, wilting time was 5 hours and additives were treated with 106 cfu/g of Lactobacillus plantarum. The silages samples were collected at 1, 2, 3, 5, 10, 20, 30, 45, and 60 days after ensiling and analyzed for the ensiling quality and characteristics of fermentation patterns. Results: Wilting treatment resulted in lower crude protein and in vitro dry matter digestibility and there were no significant differences in acid detergent fiber (ADF), total digestible nutrient (TDN), water-soluble carbohydrate (WSC), ammonia content, and pH (p>0.05). However, wilting treatment resulted in higher ADF and neutral detergent fiber content of Italian ryegrass silage (p<0.05), and the WNA treatment showed the lowest TDN and in vitro dry matter digestibility. The pH of the silage was higher in the wilting group (WNA and WA) and lower in the additive treatment group. Meanwhile, the decrease in pH occurred sharply between the 3-5th day of storage. The ammonia nitrogen content was significantly lower in the additive treatment (p<0.05), and wilting had no effect. As fermentation progressed, the lactic and acetic acid contents were increased and showed the highest content at 30 days of storage. Conclusion: The wilting treatment did not significantly improve the silage fermentation, but the inoculant treatment improved the fermentation patterns and quality of the silage. So, inoculation before ensiling is recommended when preparing high quality of Italian ryegrass silage, and when wilting, it is recommended to combine inoculation for making high quality silage.

Fungicide Spray Program to Reduce Application in Anthracnose of Strawberry (살균제 살포횟수 감소를 위한 딸기 탄저병 방제프로그램)

  • Nam, Myeong-Hyeon;Kim, Hyeon-Suk;Nam, Yun-Gyu;Peres, N.A.;Kim, Hong-Gi
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.295-301
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    • 2011
  • The effect of various fungicides on anthracnose of strawberry, caused by Colletotrichum gloeosporioides, was determined in vitro, and in greenhouse and field trials. The $EC_{50}$ values of benomyl were clearly different between two sensitive and resistant groups of isolates. Iminoctadine tris had lower $EC_{50}$ values than mancozeb and propineb as protective fungicides and the response of mancozeb, propineb and azoxystrobin was variable depending on the isolate. In the greenhouse, pre- and post- inoculation fungicide applications significantly reduced disease compared to the non-treated control. Propineb, mancozeb and azoxystrobin were effective in controlling the disease when applied prior to inoculation. Metconazole and prochloraz-Mn treatments as ergosterol biosynthesis-inhibiting fungicides had the lowest incidence of anthracnose. In the nursery field trials in 2009 and 2010, the reduced fungicide spray program provided similar levels of disease control compared to the calendar-based applications with captan. A reduced spray program based on efficacious fungicides such as prochloraz-Mn will be useful for strawberry growers and provide more options for controlling anthracnose in Korea.

ppGalNAc T1 as a Potential Novel Marker for Human Bladder Cancer

  • Ding, Ming-Xia;Wang, Hai-Feng;Wang, Jian-Song;Zhan, Hui;Zuo, Yi-Gang;Yang, De-Lin;Liu, Jing-Yu;Wang, Wei;Ke, Chang-Xing;Yan, Ru-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5653-5657
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    • 2012
  • Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.

Effects of using different roughages in the total mixed ration inoculated with or without coculture of Lactobacillus acidophilus and Bacillus subtilis on in vitro rumen fermentation and microbial population

  • Miguel, Michelle;Mamuad, Lovelia;Ramos, Sonny;Ku, Min Jung;Jeong, Chang Dae;Kim, Seon Ho;Cho, Yong Il;Lee, Sang Suk
    • Animal Bioscience
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    • v.34 no.4
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    • pp.642-651
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    • 2021
  • Objective: This study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of Lactobacillus acidophilus (L. acidophilus) and Bacillus subtilis (B. subtilis) on in vitro rumen fermentation and microbial population. Methods: Three TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of L. acidophilus and B. subtilis). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for in vitro rumen fermentation to determine total gas production, pH, ammonianitrogen (NH3-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems. Results: Changes in the fermentation (pH, lactate, acetate, propionate, and NH3-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For in vitro rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets. Conclusion: The use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of L. acidophilus and B. subtilis, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during in vitro rumen fermentation experiment.

Antitumor and Immunomodulatory Activities of Platycodon grandiflorum (길경(桔梗)이 면역활성(免疫活性) 및 항암(抗癌)에 미치는 영향)

  • Lee, Ji-Yung;Lee, Jin-Moo;Lee, Chang-Hoon;Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.4
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    • pp.10-19
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    • 2010
  • Methods: Intravenous administration of Platycodon grandiflorum was performed 2 days before tumor inoculation, then mice were killed 14 days after tumor inoculation, then number of tumor colonies were counted. Methanol extracts of Platycodon grandiflorum was added to colon26-M3.1 carcinoma cells, L5178Y-R lymphoma cells and Hela cells, and then cell growth was counted. To observe the immunomodulating effects of Platycodon grandiflorum, production of IL-6, IL-10, IL-12 and TNF-$\alpha$ were measured with ELISA assay and the cell growth of macrophage were also counted. Furthermore, antimetastatic experiment after depletion NK cells by injection of anti-asialo GM1 serum was also administered. Results: Intravenous administration of Platycodon grandiflorum significantly inhibited metastasis of colon26-M3.1 carcinoma cells. In an in vitro cytotoxicity analysis, Platycodon grandiflorum affected tumor cell growth above specific concentration. As compared with control, the production of IL-6, IL-10, IL-12 and TNF-$\alpha$ were incresed. And depletion NK cell completly abolished the inhibitory effect of metastasis. Conclusion: Platycodon grandiflorum appears to have considerable activity on immunomodulating effects and inhibit the metastasis of tumor. Further evaluation is needed for settling this.

Study on Environmental Risk Assessment for Potential Effect of Genetically Modified Nicotiana benthamiana Expressing ZGMMV Coat Protein Gene

  • Kim, Tae-Sung;Yu, Min-Su;Koh, Kong-Suk;Oh, Kyoung-Hee;Ahn, Hong-Il;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.353-359
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    • 2006
  • Transgenic Nicotiana benthamiana plants harboring the coat protein(CP) gene of Zucchini green mottle mosaic virus(ZGMMV) were chosen as a model host for the environmental risk assessment of genetically modified plants with virus resistance. This study was focused on whether new virus type may arise during serial inoculation of one point CP mutant of ZGMMV on the transgenic plants. In vitro transcripts derived from the non-functional CP mutant were inoculated onto the virus-tolerant and -susceptible transgenic N. benthamiana plants. Any notable viral symptoms that could arise on the inoculated transgenic host plants were not detected, even though the inoculation experiment was repeated a total of ten times. This result suggests that potential risk associated with the CP-expressiing transgenic plants may not be significant. However, cautions must be taken as it does not guarantee environmental safety of these CP-mediated virus-resistant plants, considering the limited number of the transgenic plants tested in this study. Further study at a larger scale is needed to evaluate the environmental risk that might be associated with the CP-mediated virus resistant plant.