• Title/Summary/Keyword: In vitro Development

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Effect of Probiotics Lactobacillus and Bifidobacterium on Gut-Derived Lipopolysaccharides and Inflammatory Cytokines: An In Vitro Study Using a Human Colonic Microbiota Model

  • Rodes, Laetitia;Khan, Afshan;Paul, Arghya;Coussa-Charley, Michael;Marinescu, Daniel;Tomaro-Duchesneau, Catherine;Shao, Wei;Kahouli, Imen;Prakash, Satya
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.518-526
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    • 2013
  • Gut-derived lipopolysaccharides (LPS) are critical to the development and progression of chronic low-grade inflammation and metabolic diseases. In this study, the effects of probiotics Lactobacillus and Bifidobacterium on gut-derived lipopolysaccharide and inflammatory cytokine concentrations were evaluated using a human colonic microbiota model. Lactobacillus reuteri, L. rhamnosus, L. plantarum, Bifidobacterium animalis, B. bifidum, B. longum, and B. longum subsp. infantis were identified from the literature for their anti-inflammatory potential. Each bacterial culture was administered daily to a human colonic microbiota model during 14 days. Colonic lipopolysaccharides, and Gram-positive and negative bacteria were quantified. RAW 264.7 macrophage cells were stimulated with supernatant from the human colonic microbiota model. Concentrations of TNF-${\alpha}$, IL-$1{\beta}$, and IL-4 cytokines were measured. Lipopolysaccharide concentrations were significantly reduced with the administration of B. bifidum ($-46.45{\pm}5.65%$), L. rhamnosus ($-30.40{\pm}5.08%$), B. longum ($-42.50{\pm}1.28%$), and B. longum subsp. infantis ($-68.85{\pm}5.32%$) (p < 0.05). Cell counts of Gram-negative and positive bacteria were distinctly affected by the probiotic administered. There was a probiotic strain-specific effect on immunomodulatory responses of RAW 264.7 macrophage cells. B. longum subsp. infantis demonstrated higher capacities to reduce TNF-${\alpha}$ concentrations ($-69.41{\pm}2.78%$; p < 0.05) and to increase IL-4 concentrations ($+16.50{\pm}0.59%$; p < 0.05). Colonic lipopolysaccharides were significantly correlated with TNF-${\alpha}$ and IL-$1{\beta}$ concentrations (p < 0.05). These findings suggest that specific probiotic bacteria, such as B. longum subsp. infantis, might decrease colonic lipopolysaccharide concentrations, which might reduce the proinflammatory tone. This study has noteworthy applications in the field of biotherapeutics for the prevention and/or treatment of inflammatory and metabolic diseases.

Effects of Equilibration Time, Precooling and Straw Loading Method on Survival of Mouse Embryos Frozen by Vitrification (생쥐 수정란의 유리화 동결보존에 있어서 동결전 처리방법에 관한 연구)

  • Gong, Il-Geun;Lee, Eun-Bong;Gang, Dae-Jin;Park, Chung-Saeng
    • Korean Journal of Animal Reproduction
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    • v.15 no.1
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    • pp.49-57
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    • 1991
  • This study was carried out to investigate the effect of equilibration time, precooling and straw loading method on the post-thaw survival rates of mouse embryos cryopreserved by vitrification method. The effect of the vitrification procedure on the damage of the embryos was also examined by the straining of nuclei using Hoechst 33342. The results obtained were summarized as follows; 1. The equilibration in Medium-1 for 10 minutes was considered to be the optimum equilibration time. Embryos equilibrated in Medium-1 for 10 minutes(81.0%) showed significantly(P<0.05) higher survival rates than those equilibrated for 5 minutes(40.0%) or 15 minutes(74.1%). 2. The survival rate of embryos cryopreserved using the double Medium-2 column(81.0%) was significantly(P<0.01) higher than that using the single Medium-2 column, whish was considered to be due to the double Medium-2 column method being more reliable for preventing contamination of diluent solution of 1M sucrose. 3. The survival rate of compacted morula stage embryos cryopreserved with the precooled and non-precooled Medium-2 was not significantly(P<0.05) different. 4. The number of blastomeres of embryos at late blastocyst stage after culture of mouse morulae for 24 to 28hours was significantly(P<0.05) decreased by freezing embryos using vitrification(53.3${\pm}$1.6 vs 41.4${\pm}$1.5), which was considered to be due to the damage of embryos during vitrification and the delay of embryo development by handling in vitro. 5. The vitrification procedure is considered to be a very simple and efficient method for cryopreservation of embryos at early developmental stage.

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Influence of Hwanhonsan Extract against Chemically Induced and Xenografted Mice Tumor (환혼산(還魂散)이 실험적(實驗的)으로 유발(誘發)한 종양(腫瘍)에 미치는 영향(影響))

  • Song, Hyo-Won;Ryu, Do-Gon;Cho, Dong-Ki;Um, Sang-Sub;Kang, Sung-Do;Go, Jeoin-Soo;Sung, Yeun-Kyung;Yun, Young-Gap;Cho, Nam-Su;Lee, Chun-Woo;Kang, Soon-Soo
    • Journal of Oriental Physiology
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    • v.14 no.2 s.20
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    • pp.229-237
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    • 1999
  • Hwanhonsan has been used for curing tumor as a Oriental medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Hwanhonsan extract against cancer, and to study some mechanisms responsible for its effect. Some kind of tumors were induced by the typical application of 3-methylcholanthrene(MCA) or by the implantation of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S180 cells) and FasII cells. Treatment of the Hwanhonsan extract(daily 1 mg/mouse, i.p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 hrs. Against squamous cell carcinoma induced by MCA, Hwanhonsan decreased. not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Hwanhonsan also significantly suppressed the development of 3LL cells and S180 cells implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Hwanhonsan extract into TBM. However, when tumor was induced by FsaII cells implantation, the growth of implanted cells in mice was delayed by the water extract of Hwanhonsan until 7 days and then rapid growth ensued. In vitro treatment of Hwanhonsan extract had no inhibitory effect on the tumor induced by some kind of cell lines such as A431 cells strain but it significantly inhibited the proliferation of 3LL cells, S180 cells. These results suggested that Hwanhonsan extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells.

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Pancreatic Lipase Inhibitory and Antioxidant Activities of Zingiber officinale Extracts (생강 추출물의 pancreatic lipase 저해 및 항산화 활성)

  • Bae, Jong-Sup;Kim, Tae-Hoon
    • Food Science and Preservation
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    • v.18 no.3
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    • pp.390-396
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    • 2011
  • Ginger (Zingiber officinale) is a well-known herb that is widely consumed as spice for the flavoring of foods. As part of our continuing search for bioactive materials, the in vitro pancreatic lipase inhibition and antioxidant properties of an aqueous ethanolic extract of Z. officinale were investigated. The total phenolic content was determined using a spectrophotometric method. The antioxidant efficacies of the extract was studied with radical scavenging assays using DPPH and $ABTS^+$ radicals. Further more, the antiobesity effect of the extract was evaluated by porcine pancreatic lipase assay. In particularly, the pancreatic lipase inhibitory activity of the ethyl acetate (EtOAc)-soluble portion from Z. officinale was significantly higher than that of the other solvent-soluble portions. The results suggest that Z. officinale may have therapeutic potential that may be useful in development of an anti-obesity agent or its precursors.

Development of Solid Self-nanoemulsifying Drug Delivery Systems of Ticagrelor Using Porous Carriers (다공성의 캐리어를 이용한 티카그렐러 함유 고형의 자가 나노유화 약물전달시스템 개발)

  • Choi, Hyung Joo;Kim, Kyeong Soo
    • Journal of Life Science
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    • v.31 no.5
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    • pp.502-510
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    • 2021
  • The objective of this study was to develop a novel ticagrelor-loaded self-nanoemulsifying drug delivery system with an enhanced solubility and dissolution rate. Numerous oils and surfactants were screened, then medium chain triglyceride (MCT) oil and the surfactants polyoxyethylene sorbitan monooleate (Tween 80) and Labrafil M1944CS were selected for the preparation of the ticagrelor-loaded self-nanoemulsifying drug delivery system. A pseudo-ternary phase diagram was constructed to detect the nanoemulsion region. Of the various formulations tested, the liquid SNEDDS, composed of MCT (oil), Tween 80 (surfactant), and Labrafil M1944CS (cosurfactant) at a weight ratio of 20/70/10 produced the smallest emulsion droplet size (around 20.56±0.70 nm). Then, particle size, polydispersity, and zeta potential were measured using drugs containing liquid SNEDDS. The selected ticagrelor-loaded liquid SNEDDS was spray-dried to convert it into a ticagrelor-loaded solid SNEDDS with a suitable inert carrier, such as silicon dioxide, calcium silicate, or magnesium aluminometasilicate. The solid SNEDDS was characterized by scanning electron microscopy, transmission electron microscopy, and in vitro dissolution studies. SEM, PXRD, and DSC results suggested that amorphous ticagrelor was present in the solid SNEDDS. Also, the solid SNEDDS significantly increased the dissolution rate of ticagrelor. In particular, the emulsion particle size and the polydispersity index of the solid SNEDDS using silicon dioxide (SS1) as a carrier was the smallest among the evaluated solid SNEDDS, and the flowability and compressibility result of the SS1 was the most suitable for the manufacturing of solid dosage forms. Therefore, solid SNEDDS using silicon dioxide (SS1) could be a potential nano-sized drug delivery system for the poorly water-soluble drug ticagrelor.

Application of Lactococcus lactis HY7803 into Soybean Fermentation for Production of Glutamic Acid (글루탐산 생산을 위한 Lactococcus lactis HY7803 균주의 대두 발효 적용)

  • Jungmin Lee;Sojeong Heo;Jihoon Choi;Eunji Pyo;Myounghee Lee;Sangick Shin;Jaehwan Lee;Junglyoul Lee;Do-Won Jeong
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.53-58
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    • 2023
  • The glutamic acid producing strain for development savory taste enhancing foods was identified the possibility through application into soybean fermentation. To check the effects on glutamic acid production during soybean fermentation, Lactococcus lactis HY7803 was introduced as a starter. The soybean samples were analyzed on days 0, 7, 14 and 21. The numbers of bacteria decreased gradually, while the content of amino-type nitrogen increased during fermentation in the soybean with L. lactis HY7803. Glutamic acid content in soybeans with L. lactis HY7803 increased from 114.99 ± 9.37 pmol/ul on day 0 to 138.14 ± 1.76 pmol/ul on day 21, showing an overall higher amino acid content than soybeans without L. lactis HY7803 and similar content to soybeans with Aspergillus oryzae SNU-G. It was clearly distinguished through principal component analysis. Consequently, our results indicate that L. lactis HY7803 is available as a fungus replacement and may be a good starter strain for enhancing savory taste in vitro as well as soybean fermentation.

Studies on Offspring Production by Transfer of Bisected Demi-Embryos in Mice (생쥐에 있어서 양분분할구(兩分分割球) 이식(移植)에 의한 새끼생산(生産)에 관한 연구(硏究))

  • Suh, Tae Kwang;Park, Hang Kyun
    • Current Research on Agriculture and Life Sciences
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    • v.6
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    • pp.145-156
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    • 1988
  • This study was conducted to investigate the effect of embryo stage on bisection rates of embryos, on development of separated demi-embryos and on subsequent development to full term following transfer of demi-embryos to recipients. The results obtained in this study are summarized as follows. The 2-, 4-, 8-cell and morula embryos were mostly obtained at 36-42, 48-54, 62-66 and 72-78 hours after injection of hCG, respectively, and the average number of embryos recovered per head was 17.0. The bisection rate of 8-cell embryos was 83.6%, which was significantly higher than that of morula embryos, 65.5%. But the development of morula demi-embryos to blastocyst after in-vitro culture was better than that of 8-cell demi-embryos and the rate was 76.5% and 60.9%, respectively. When the 2, 10 and 15 cultured demi-embryos or intact embryos were transferred to each recipient mouse, respectively. the highest pregnancy rate was obtained when 15 embryos were transferred. And the overall offspring production rate of intact embryos was higher than that of demi-embryos.

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Effects of Gunleetang Gagambang Extract on Antitumoral Immunological Response and the Side Effect Induced by Antitumoral Agents (군리탕가감방(君理湯加減方)이 항종양(抗腫瘍) 면역반응(免疫反應)과 항암제로 유발(誘發)한 부작용(副作用)에 미치는 영향(影響))

  • Yui, Guyng-Tea;Moon, Suk-Jae;Moon, Goo;Won, Jin-Hee
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.71-87
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    • 1998
  • Even though appropriate immune response is necessary for the survival of the individual, excessive or insufficient immune Response might cause autoimmune or allergic disease. So the immune response must be controlled to the degree that is beneficial for the well being of the individual. This study was undertaken to know the effects of Gunleetang Gagambang on the immune system of the mouse. Gunleetang Gagambang has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Gunleetang Gagambang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene(MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S180 cells). Treatment of the Gunleetang Gagambang on water-extract(dailly 1mg/mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Gunleetang Gagambang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice(TBM). Gunleetang Gagambang on also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurrence-frequency and their size. and some developed tumors were regressed by the continuous treatment of Gunleetang Gagambang extract into TBM. In vitro, treatment of Gunleetang Gagambang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Gunleetang Gagambang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Gunleetang Gagambang administration to mice enhanced NK cells activities. These results demonstrated that Gunleetang Gagambang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Gunleetang Gagambang might be chiefly due to nonspecitie enhancement of NK cell activities and cell-mediated immune responses.

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Morphogenetic Responses according to Media and Growth Regulators in vitro Culture of Eight Populus spp. (배지(培地) 및 생장(生長)호르몬에 따른 기내배양(器內培養)된 포푸라속(屬) 8개(個) 수종(樹種)의 기관형성반응(器官形成反應))

  • Kim, Chi Moon;Kwon, Ki Won;Moon, Heung Kyu;Lee, Jae Soon
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.205-212
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    • 1987
  • To examine the morphogenetic response, stem segments of 8 Populus spp. and 3 different explants of P. nigra var. italica were cultured on MS (Murashige and Skoog 1962) and WPM (Woody Plant Medium) medium containing various phytohormones. The results obtained were as follows: 1. Shoot regeneration and development from stem segment of 8 Populus spp. showed a quite difference according to the section and the species. All of the species of Leuce and Tacamahaca section did not form adventitious buds, while most of explants showed axillary or dormant bud elongation after 4 weeks. But P. nigra var. italica of Aigeiros section showed a successful adventitious bud formation (mean 5.4 buds per explant). 2. Leaf, petiole, and internode segment of P. nigra var. italica showed a quite differences according to media and ex plants upon the morphogenetic response. Adventitious bud formation from leaf was more abundant and readily initiated on the abaxial side than on the adaxial side. Mean number of 103 adventitious buds per explant was obtained from abaxial side of leaf segment cultured on WPM medium containing $0.2mg/{\ell}$ BAP for 5 weeks. 3. 2,4-D (2,4-dichlorophenoxy acetic acid) supplemented to media appeared to be negative upon the adventitious bud formation of P. nigra var. italica, while it promoted callus formation from all explants. Especially, NAA (${\alpha}$-naphtalene acetic acid) or NAA combination with BAP (6-benzylaminopurine) promoted root regeneration from the all explant of P. nigra var. italica in this study.

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Systematic Identification of Hepatocellular Proteins Interacting with NS5A of the Hepatitis C Virus

  • Ahn, Ji-Won;Chung, Kyung-Sook;Kim, Dong-Uk;Won, Mi-Sun;Kim, Li-La;Kim, Kyung-Shin;Nam, Mi-Young;Choi, Shin-Jung;Kim, Hyoung-Chin;Yoon, Mi-Chung;Chae, Suhn-Kee;Hoe, Kwang-Lae
    • BMB Reports
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    • v.37 no.6
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    • pp.741-748
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    • 2004
  • The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.