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The Effect of Storage Temperature on the Viability and Bioluminescence of Photobacterium phosphoreum (저장 온도에 따른 Photobacterium phosphoreum의 Viability와 Bioluminescene)

  • Kim, Hyeon-Suk;Jeong, Seong-Je;Jeong, Gye-Hun;Jeon, Eok-Han
    • KSBB Journal
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    • v.14 no.5
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    • pp.523-527
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    • 1999
  • A naturally luminescent bacterium, Photobacterium phosphoreum was stored in 2.5% NaCl solution at 2$0^{\circ}C$, 4$^{\circ}C$, -2$0^{\circ}C$ and -7$0^{\circ}C$ for 30 days. In vivo luminescence and concentrations of total and culturable cells were determined by luminometer, spectrophotometer and dilution plate counting, respectively. When stored at 4$^{\circ}C$ and 2$0^{\circ}C$, concentrations of cells were rapidly decreased as a result of cell lysis, leading to adrop in turbidity and cultured counts. The bioluminescence of cells stored at 4$^{\circ}C$ was maintained until 12 days while those of cells starved at other temperatures decreased to background level within 3 days. Following incubation of stored cells in fresh liquid medium, activities of viable cells increased throughout storage period excepting cells stored at 2$0^{\circ}C$. Changes in bioluminescence intensity following addition of 2.5% NaCl solution markedly showed in cells stored at -2$0^{\circ}C$ and -7$0^{\circ}C$ and increased to maximum 8 fold.

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Inorganic sulfur reduces the motility and invasion of MDA-MB-231 human breast cancer cells

  • Kim, Jin-Joo;Ha, Hwa-Ae;Kim, Hee-Sun;Kim, Woo-Kyoung
    • Nutrition Research and Practice
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    • v.5 no.5
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    • pp.375-380
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    • 2011
  • This study investigated the effects of inorganic sulfur on metastasis in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured in the absence or presence of various concentrations (12.5, 25, or 50 ${\mu}mol$/L) of inorganic sulfur. Cell motility, invasion, and the activity and mRNA expression of matrix metalloproteases (MMPs) were examined. Numbers of viable MDA-MB-231 cells did not differ by inorganic sulfur treatment from 0 to 50 ${\mu}mol$/L within 48 h. Inorganic sulfur significantly decreased cell motility and invasion in the MDA-MB-231 cells in a dose-dependent manner (P<0.05), as determined using a Boyden chamber assay and a Matrigel chamber. The activities of MMP-2 and MMP-9 were significantly reduced by inorganic sulfur in a dose-dependent manner (P<0.05). The inorganic sulfur also significantly inhibited MMP-2 and MMP-9 expression in the cells (P<0.05). These data suggest that inorganic sulfur can suppress cancer cell motility and invasion by inhibiting MMP-2 and MMP-9 activity and gene expression in MDA-MB-231 cells.

Fermentation Characteristics of Set-Type Yoghurt from Milk Added with Mugwort Extract (쑥 추출물이 첨가된 Set-Type Yoghurt의 발효 특성)

  • 배인휴;홍기룡;오동환;박정로;최성희
    • Food Science of Animal Resources
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    • v.20 no.1
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    • pp.21-29
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    • 2000
  • This study was carried out to investigate the fermentation characteristics and storage of set-type yoghurt added mugwort extracts(AME) such as pH, growth of lactic acid bacteria, number of viable cells, viscosity, and sensory characteristics during 24 hours fermentation and 15 days storage. Addition of mugwort extracts was grown rapidly of lactic acid bacteria rather than that of control and also 4 or 8% AME groups were grown similar to control. The drop of AME pH of broth was less compared with control during incubation of lactic acid bacteria. The growth of lactic acid bacteria during incubation of AME yoghurt was not different of viable cell count between AME group and control in beginning time, but the viable cell count of AME groups were increased depended opon addition quantity of AME in ending time. Addition of mugwort extracts was not affect on pH change during yoghurt fermentation and increased a lactic acid bacteria number as well as no effect of yoghurt fermentation in ending time. The viscosity of yoghurt was almost not changed 3 hours after yoghurt mix and increased rapidly 6 hours after yoghurt mix. Although control and 0.5% AME group showed maximum viscosity at 18 hours of fermentation, 1 and 2% AME group showed linear increase until 24 hours of fermentation. Mugwort did not affect pH and viable cel number of lactic acid bacteria during 15 days storage 24 hours after fermentation. Sensory evaluation of the AME yoghurt showed that flavour, texture and acid taste were not affected by addition of mugwort. However, the appearance and taste were dropped by addition of mugwort.

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Chitinase을 생산하는 곤충병원미생물 Metarhizium anisopliae HY-2(KCTC 0156BP)의 토양해충 생물검정

  • Seo, Eun-Yeong;Son, Gwang-Hui;Sin, Dong-Ha;Kim, Gi-Deok;Park, Du-Sang;Park, Ho-Yong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.469-472
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    • 2002
  • Solid state fermentation was performed for the production of entomopathogenic fungus Metarhizium anisopliae HY-2 using wheat bran media containing rice bran. Fungal growth in a solid state fermentation system was estimated by viable cell count, spore count, and mycelial biomass. It was used chemical method measuring N-acetyl-glucosamine (chitin) content for estimating of mycelial biomass. In static flask culture, viable cell reached 2.40 ${\times}$ $10^8$ cfu/g at 23 days of culture at $27^{\circ}C$ and then mycelial biomass was 41.59 mg/g. Specific growth rate(${\mu}$ max) was 0.0418 $h^{-1}$ between 3 and 9 days when estimated by viable cell count and was 0.00976 $h^{-1}$ between 9 and 17 days when N-acetylglucosamine content was measured. Viable cells reached 1.12 ${\times}$ $10^8$ cfu/g in polypropylene-bag at 28 days of culture at $27^{\circ}C$. Formulated microbial pesticide containing M. anisopliae HY-2 were tested their bio-activity against Chestnut Brown Chafer (Adoretus tenuimaculatus). The protection rate of the liquid culture showed 13 ${\sim}$ 26 % with 1st to 3rd instar, and spore suspension of M. anisopliae HY-2 showed 56 ${\sim}$ 64%. Conidia produced by large scale solid-state fermentation showed 20 ${\sim}$ 27 % activity 60 ${\sim}$ 64 % with M. anisopliae HY-2.

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Improvement of Shelf-life of Yakju by Membrane Filtration (Membrane Filtration에 의한 약주의 저장성 증진)

  • Kang, Mi-Young;Park, Young-Seo;Mok, Chul-Kyoon;Chang, Hak-Gil
    • Korean Journal of Food Science and Technology
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    • v.30 no.5
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    • pp.1134-1139
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    • 1998
  • Quality changes of filter-sterilized Yakju were examined by filtration of Yakju through membranes followed by storing at $25^{\circ}C$ for 50 days. To evaluate quality changes of filter-sterilized Yakju, pH, titratable acidity, turbidity, and viable cell numbers of total bacteria, lactic acid bacteria, and yeast were measured. Titratable acidity, turbidity and viable cell numbers of non-sterilized Yakju increased, but pH profile decreased during the storage. In filter-sterilized Yakju, titratable acidity and turbidity did not change, while viable cells of total bacteria, lactic acid bacteria, and yeast were not detected during the storage. Addition of chitosan at the concentration of 0.1% (w/v) decreased the viable cell numbers significantly showing similar pH and titratable acidity profiles with non-sterilized Yakju.

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Development of Reverse Transcriptase-Polymerase Chain Reaction of fimA Gene to Detect Viable Salmonella in Milk (우유 내 활력있는 Salmonella를 검출하기 위한 fimA 유전자의 역전사중합효소 연쇄반응의 개발)

  • Choi, S.H.;Lee, S.B.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.841-848
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    • 2004
  • Rapid detection of viable Salmonella in pasteurized milk is important to protect public health from food poisoning. Reverse transcriptase-polymerase chain reaction(RT-PCR) is recognized as a molecular genetical method to differentiate between live and dead bacteria The RT-PCR in this study was designed to detect specifically viable Salmonella in milk by using the primers whose nucleotide sequences were determined based on fimA gene which encodes the submit of type 1 fimbriae. Treatment of RNA preparation with RNase-free DNase was adequate enough to destroy DNA, which may otherwise be amplified in the RT PCR Seven strains of Salmonella were detected in the RT-PCR but Escherichia coli, Shigella sonnei, Citrobacter freundii, and Klebsiella pneumoniae were not. $10^7/ml$ and $10^6/ml$ of dead Salmonella which were heat-treated in milk were detectable by using the RT-PCR but $10^5{\sim}10/ml$ of the dead bacteria were not. The sensitivity of the RT-PCR in detecting viable Salmonella was 100 cells/ml.

Artificial gametes from stem cells

  • Moreno, Inmaculada;Miguez-Forjan, Jose Manuel;Simon, Carlos
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.2
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    • pp.33-44
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    • 2015
  • The generation of artificial gametes is a real challenge for the scientific community today. In vitro development of human eggs and sperm will pave the way for the understanding of the complex process of human gametogenesis and will provide with human gametes for the study of infertility and the onset of some inherited disorders. However, the great promise of artificial gametes resides in their future application on reproductive treatments for all these people wishing to have genetically related children and for which gamete donation is now their unique option of parenthood. This is the case of infertile patients devoid of suitable gametes, same sex couples, singles and those fertile couples in a high risk of transmitting serious diseases to their progeny. In the search of the best method to obtain artificial gametes, many researchers have successfully obtained human germ cell-like cells from stem cells at different stages of differentiation. In the near future, this field will evolve to new methods providing not only viable but also functional and safe artificial germ cells. These artificial sperm and eggs should be able to recapitulate all the genetic and epigenetic processes needed for the correct gametogenesis, fertilization and embryogenesis leading to the birth of a healthy and fertile newborn.

Effects of Daturae Flos and Daturae Semen Extract on the Immunocyte Response in Mice (만타라화(曼陀羅花) 및 만타라자(曼陀羅子) 수추출물(水抽出物)이 마우스의 면역세포기능(免疫細胞機能)에 미치는 영향(影響))

  • Ko, Woon-Che;Song, Ho-Joon;Shin, Min-Kyo
    • Korean Journal of Pharmacognosy
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    • v.21 no.4
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    • pp.307-316
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    • 1990
  • This study was undertaken to test the effects of Daturae Flos(DF) and Daturae Semen(DS) on the cellular and humoral immune responses, and the functions of the cells involved in immunoinflammation. Both extracts decreased the activity of superoxide dismutase, and the decrease was greater in the mouse group which was treated with DS. Both extracts decreased the phagocytic activity as measured by assessing the number of the latex particle within the phagocyte after incubation of peritoneal macrophages with fluorochrome-labelled latex particle and decreased natural killer cell activity as measured by enumerating the viable YAC-1 cells after treatment of target cells with splenic natural killer cells. Both extracts also decreased the cell-mediated immunity in vivo as assessed by measuring the ear thickness after sensitization and challenge with dinitrofluorobenzene, however, had no effects on the humoral immune responses as measured by checking hemolysin and hemagglutinin titers after immunization with sheep red blood cells(SRBC). Extracts of Semen caused decrease in the number of rosette forming cells between the splenic cells and SRBC. The results of this study suggested that both Daturae extracts could depress the immunoinflammation by affecting the various cell types involved in inflammation.

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Globotriaosylceramide (Gb3) content in HeLa cells is correlated to Shiga toxin-induced cytotoxicity and Gb3 synthase expression

  • Shin, In-Sun;Ishii, Satoshi;Shin, Jong-Seo;Sung, Kyong-Il;Park, Byung-Sung;Jang, Hyun-Yong;Kim, Byong-Wan
    • BMB Reports
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    • v.42 no.5
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    • pp.310-314
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    • 2009
  • Globotriaosylceramide (Gb3) and globotetraosylceramide (Gb4) are the proposed functional receptors for Shiga toxin (Stx). To elucidate the effect of Gb3 content on Stx-induced cytotoxicity in HeLa cells, we cloned HeLa cells and determined the correlation between glycolipids content and Stx-induced cytotoxicity. The 29 HeLa cell clone (HLCC) lines used showed a wide range of sensitivity to Stx, compared to Gb3-rich cells which were more sensitive, showing as little as 20% viability to 100 pg/ml Stx. In contrast, Gb3-deficient cells proved resistant as they were more than 80% viable to 100 ng/ml Stx. Gb3 content in the HLCC lines corresponded with Stxs-induced cytotoxicity as well as Gb3 synthase expression, but no correlation with Gb4 content was noted. These data show that Gb3 content, which is regulated by the expression of Gb3 synthase, determines the sensitivity of HeLa cells toward Stx.

Methodologies for Cryopreservation of Mammalian Germline Cells and Tissues

  • Karmakar, Polash Chandra;Jung, Sang-Eun;Ryu, Buom-Yong
    • Reproductive and Developmental Biology
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    • v.41 no.2
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    • pp.41-50
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    • 2017
  • Until today, success in germline cells and tissue cryopreservation is limited mainly due to the poor understanding of the complex physiological processes can lead to cell damage during cryopreservation. Germline cells, from both male and female, have unique ability to differentiate into one or more cell lines and thus it becomes a crucial point to store them in subzero temperature with the minimal damage of their functional properties and maximum recovery of unchanged and viable cells when thawed. In the past three decades, a vast research has been performed using various different animal models which in fact have led to development of new methodologies and optimization of older one. However, successful use of animal model has provided the opportunity in research with human germline cells and tissues preservation, but not in all the cases. Therefore, the use of new cryo-protective chemicals and modified protocols have been often found in different groups of researchers based on the types, physical structures, utility and animal species of the specimens to be cryopreserved. This review discusses about the basics of different types of cryopreservation methodologies and commonly used optimized protocols and cryoprotectants for germline cells and tissues preservation.