• Title/Summary/Keyword: phosphate-free medium

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Production of Mushroom Mycelium (Agaricus campestris) in Shaking Culture (진탕배양법(振?培養法)에 의한 양송이 균사체(菌絲體)의 생산(生産))

  • Lee, Jeong-Sook;Lee, Su-Rae;Yu, Tai-Jong
    • Korean Journal of Food Science and Technology
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    • v.7 no.1
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    • pp.22-29
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    • 1975
  • Conditions for submerged culture of Agaricus campestris var. bisporus and the chemical composition of its mycelium were investigated. In shaking culture with TGY basal medium at $27{\sim}30^{\circ}C$, pH tended to increase upon culture period, mycelial growth was the highest on 12th day, with relatively high nitrogen content of 7% and sugar in the medium disappeared almost at the end of culture period. As a nitrogen source, ammonium phosphate (dibasic) gave relatively high mycelial yield and the addition of yeast extract gave rise to better results. As a carbon source, glucose was the best, fructose, maltose, lactose and sucrose gave the same results, and soluble starch was utilized slightly. Mushroom mycelium contained 48% of protein, 8 free amino acids including arginine, histidine, lysine, isoleucine, leucine, phenylalanine, proline, tyrosine and its protein consisted of most essential amino acids, with relatively high contents of lysine and threonine. Therefore, mushroom mycelium deserves to be a high quality protein food.

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Establishment of Suspension Culture System to Induce Somatic Embryo in Oplopanax elatus Nakai (현탁배양을 통한 땃두릅나무의 체세포배 유도)

  • Kim, Hee Young;Seong, Eun Soo;Lee, Jae Geun;Yoo, Ji Hye;Hwang, In Seong;Kim, Myong Jo;Lim, Jung Dae;Kim, Na Young;Yu, Chang Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.461-465
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    • 2012
  • This study was carried out to select the appropriate medium (especially, carbon and nitrogen source, potassium phosphate and pH) for somatic embryogenesis in order to develop the rapid mass production system in suspension culture of Oplopanax elatus Nakai. Direct somatic embryos were obtained from root explants in the hormone free suspension culture (MS). Combination of $NH_4NO_3$ and $KNO_3$ at the ratio of 1650 (mg/${\ell}$) : 1900 (mg/${\ell}$) obtained the better result to produce somatic embryo in suspension culture. MS medium supplemented with $170mg/{\ell}$ $KH_2PO_4$. The addition of 1 and 3% sucrose was effective for formation of embryogenic callus. Therefore, this report will be helped to improve the establishment for suspension culture in Oplopanax elatus Nakai.

Voltage Dependent N Type Calcium Channel in Mouse Egg Fertilization

  • Eum, Jin Hee;Park, Miseon;Yoon, Jung Ah;Yoon, Sook Young
    • Development and Reproduction
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    • v.24 no.4
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    • pp.297-306
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    • 2020
  • Repetitive changes in the intracellular calcium concentration ([Ca2+]i) triggers egg activation, including cortical granule exocytosis, resumption of second meiosis, block to polyspermy, and initiating embryonic development. [Ca2+]i oscillations that continue for several hours, are required for the early events of egg activation and possibly connected to further development to the blastocyst stage. The sources of Ca2+ ion elevation during [Ca2+]i oscillations are Ca2+ release from endoplasmic reticulum through inositol 1,4,5 tri-phosphate receptor and Ca2+ ion influx through Ca2+ channel on the plasma membrane. Ca2+ channels have been characterized into voltage-dependent Ca2+ channels (VDCCs), ligand-gated Ca2+ channel, and leak-channel. VDCCs expressed on muscle cell or neuron is specified into L, T, N, P, Q, and R type VDCs by their activation threshold or their sensitivity to peptide toxins isolated from cone snails and spiders. The present study was aimed to investigate the localization pattern of N and P/Q type voltage-dependent calcium channels in mouse eggs and the role in fertilization. [Ca2+]i oscillation was observed in a Ca2+ contained medium with sperm factor or adenophostin A injection but disappeared in Ca2+ free medium. Ca2+ influx was decreased by Lat A. N-VDCC specific inhibitor, ω-Conotoxin CVIIA induced abnormal [Ca2+]i oscillation profiles in SrCl2 treatment. N or P/Q type VDC were distributed on the plasma membrane in cortical cluster form, not in the cytoplasm. Ca2+ influx is essential for [Ca2+]i oscillation during mammalian fertilization. This Ca2+ influx might be controlled through the N or P/Q type VDCCs. Abnormal VDCCs expression of eggs could be tested in fertilization failure or low fertilization eggs in subfertility women.

Production and Characterization of an Anti-Angiogenic Agent front Saccharomyces cerevisiae K-7

  • Jeong, Seung-Chan;Lee, Dae-Hyoung;Lee, Jong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1904-1911
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    • 2006
  • The cell-free extracts of 250 yeasts were screened for their in vitro anti-angiogenic activity, to develop a new cancer metastasis inhibitor. Saccharomyces cerevisiae K-7 was selected as the producer of the anti-angiogenic agent, because it had the highest anti-angiogenic activity. The anti-angiogenic agent was produced maximally from hydrolysates of Saccharomyces cerevisiae K-7, when the yeast was cultured in yeast extract-peptone-dextrose medium at 30$^{\circ}C$ for 24 h, and cell-free extracts were than digested with pepsin for 4 h at 37$^{\circ}C$. The anti-angiogenic agent was further purified by ultrafiltration, Sephadex G-25 gel permeation chromatography and reverse-phase HPLC, and the anti-angiogenic activity of the final purified preparation was 72.7% at 10 $\mu$M/egg. The purified anti-angiogenic agent was found to originate from the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) molecule of Saccharomyces cerevisiae K-7, and its peptide sequence was Val-Ser-Trp-Tyr-Asp-Asn-Glu-Tyr-Gly-Tyr-Ser-Thr-Arg-Val-Val-Asp. In the MTT assay, the shape of the HT-l 080 cell was clearly changed to a circular type at 0.2 mM purified anti-angiogenic agent. This result indicated that the growth of the HT-I080 cell was significantly inhibited at 0.2 mM of the purified anti-angiogenic agent. The MMP activity of the treated HT-l080 cells was not affected, evidenced by the gelatin zymography, indicating that the anti-angiogenic mechanism of the purified anti-angiogenic agent is not mediated through MMP activity.

A NOTE ON THE REMOVAL OF PHYTATE IN SOYBEAN MEAL USING Aspergillus usami

  • Ilyas, A.;Hirabayasi, M.;Matsui, T.;Yano, H.;Yano, F.;Kikishima, T.;Takebe, M.;Hayakawa, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.2
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    • pp.135-138
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    • 1995
  • Soybean meal was fermented by Aspergillus usami in order to reduce phytate content. Aflatoxin B1 was not detected in the fermented soybean meal. The contents of crude protein, crude fiber, ether extract and crude ash were slightly increased following fermentation with a concomitant reduction in nitrogen free extract. Though the fermentation partly degraded proteins in the soybean meal, there was small difference in amino acid composition between the soybean meal and the fermented soybean meal. The results showed that the fermentation did not affect nutritional value of protein in soybean meal. Approximately 55% of phosphorus extracted by trichloroacetic acid was inositol hexaphosphate (phytate) in the soybean meal. The content of inositol tetra to hexaphosphates was not detected in the fermented soybean meal. These results indicated that the fermentation almost completely eliminated phytate in soybean meal. Phytase activity was not detected in the unfermented soybean meal. However, the enzyme activity in the fermented soybean meal was 167.7 U/g. When the fermented soybean meal in supplemented in formula feeds, phytase in the fermented soybean meal might partly degrade the phytate in other ingredients in the digestive tract. The fermented soybean meal is possibly used as a phytate-free protein source of feed, which contains high available phosphorus.

Studies on the Glucose Isomerizing Enzyme -Part III The Effects of Metal Ions on the Reaction of K-17 Strain's Glucose Isomerizing Enzyme- (포도당(葡萄糖) 이성화(異性化) 효소(效素)에 관(關)한 연구(硏究) -제3보(第三報) 분리(分離) 선정(選定)된 균주(菌株) 효소(酵素)의 작용(作用)에 미치는 금속염(金屬鹽)의 영향(影響)에 대(對)해서-)

  • Seu, J.H.;Kim, C.K.;Ki, W.K.;Rhee, I.K.;Kwon, T.J.;Woo, D.L.
    • Applied Biological Chemistry
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    • v.11
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    • pp.55-61
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    • 1969
  • This K-17 strain was not absolutely requiring xylose as an inducer for enzyme formations. The most activity of this enzyme was lost when treated at $75^{\circ}C$. for 30 hours but was not influenced at $70^{\circ}C$. for 70 hours of treatment. The activity of this enzyme was increased by the addition of magnesium ions or cobalt ions in the reaction system. In the studies, we found that the magnesium ions simply activate the enzyme reaction and the cobalt ions do not but protect the enzyme from heat inactvation. And it was also found the phosphate buffer solution was very suitable as glucose dissolving solvent on the enzyme reaction. The mixed carbon source medium containing glucose, fructose, sorbitol, xylose and sucrose was more favorable for enzyme production then a sole carbon source containing medium on the shaking culture method.

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Regeneration of adventitious root from Calystegia soldanella L. in Jeju island and mass proliferation method using bioreactor system (제주지역 갯메꽃(Calystegia soldanella L.) 유래 부정근 재분화 및 생물반응기 시스템 이용 대량증식법)

  • Jong-Du Lee;Eunbi Jang;Weon-Jong Yoon;Yong-Hwan Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.37-37
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    • 2021
  • Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

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Aldose Reductase Inhibitor Fidarestat as a Promising Drug Targeting Autophagy in Colorectal Carcinoma: a Pilot Study

  • Pandey, Saumya
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.4981-4985
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    • 2015
  • Background: Colorectal cancer (CRC) is a leading cause of morbidity and mortality worldwide. Targeting autophagic cell death is emerging as a novel strategy in cancer chemotherapy. Aldose reductase (AR) catalyzes the rate limiting step of the polyol pathway of glucose metabolism; besides reducing glucose to sorbitol, AR reduces lipid peroxidation-derived aldehydes and their glutathione conjugates. A complex interplay between autophagic cell death and/or survival may in turn govern tumor metastasis. This exploratory study aimed to investigate the potential role of AR inhibition using a novel inhibitor Fidarestat in the regulation of autophagy in CRC cells. Materials and Methods: For glucose depletion (GD), HT-29 and SW480 CRC cells were rinsed with glucose-free RPMI-1640, followed by incubation in GD medium +/- Fidarestat ($10{\mu}M$). Proteins were extracted by a RIPA-method followed by Western blotting ($35-50{\mu}g$ of protein; n=3). Results: Autophagic regulatory markers, primarily, microtubule associated protein light chain (LC) 3, autophagy-related gene (ATG) 5, ATG 7 and Beclin-1 were expressed in CRC cells; glyceraldehyde-3 phosphate dehydrogenase (GAPDH) was used as an internal reference. LC3 II (14 kDa) expression was relatively high compared to LC3A/B I levels in both CRC cell lines, suggesting occurrence of autophagy. Expression of non-autophagic markers, high mobility group box (HMG)-1 and Bcl-2, was comparatively low. Conclusions: GD +/- ARI induced autophagy in HT-29 and SW-480 cells, thereby implicating Fidarestat as a promising therapeutic agent for colorectal cancer; future studies with more potent ARIs are warranted to fully dissect the molecular regulatory networks for autophagy in colorectal carcinoma.

Effect of Lysophosphatidic Acid on Proliferation and Differentiation of Rat Skeletal Myoblasts in Culture

  • Kwon, Min-Seong;Cho
    • Animal cells and systems
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    • v.1 no.4
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    • pp.641-646
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    • 1997
  • Lysophosphatidic acid (LPA; 1-acyl-glycerol-3-phosphate) has been known as an intercellular phospholipid messenger with a wide range of biological activities. In this study, the effect of LPA on both the proliferation and differentiation of rat E63 myoblasts has been investigated. In the serum-free Insulin-Transferrin-Selenium (ITS) media, the proliferation of E63 cells was largely restricted. Addition of LPA into the ITS media strongly promoted the cell proliferation and resulted in two to four fold increase of cell number. Furthermore, it appeared to increase the percent fusion in a dose-dependent manner up to 15 ug/ml. The synthesis of myosin heavy chain (MHC) was increased by LPA as well. These results indicate that LPA is able to promote both cell proliferation and differentiation in rat E63 myoblasts. Suramin, known to have uncoupling activity on growth factor-receptor interaction, was tested for antagonistic activity in myoblast proliferation and differentiation. Myoblasts grown in the ITS medium containing LPA were able to proliferate well even in the presence high concentration of suramin whereas myoblast differentiation was completely blocked by 30 ug/ml of suramin. The inhibitory effect of suramin on the myoblast differentiation was completely reversible by removing the suramin. This result indicates that the intracellular signaling pathway of LPA leading to cell proliferation might be distinct from that leading to cell differentiation on E63 myoblasts. Also, the antagonistic effect of suramin suggests that the differentiation activity elicited by LPA might be mediated by a specific G protein-coupled receptor.

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The Effects of Kimchi on Hematological and Immunological Parameters in vivo and in vitro (In vivo와 in vitro에서 김치가 혈액성상과 면역세포배양에 미치는 영향)

  • 송영선;김미정;권명자;송영옥;이은경;윤현주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1208-1214
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    • 1997
  • This study was designed to know the effect of kimchi on the hematological and immunological parameters in vivo and in vitro, respectively. To study the effects of kimchi on the hematological parameters, rats(S.D., male) were divided into 4 groups and fed diets containing of 3%, 5% and 10% kimchi or kimchi free diet(control) for 6 weeks. The results of CBC(complete blood cell) tests obtained from the bloods of rates were as follows ; In 10% kimchi group, the level of WBC(white blood cells), RBC(red blood cells), Hgb(hemoglobin), Hct(hematocrit) were increased significantly than those of control group(p<0.05). MCV(mean corpuscular volume), one of the red cell indices, was also increased significantly in the animals fed 10% kimchi(p<0.05). RDW(Red cell distritution width) and PCT(plateletcrit) was lowest in 10% kimchi group(p<0.05). To examine the effects of kimchi on immune cell growth in vitro, three types of mouse immune cells-spleen cells, bone marrow cells, thymus cells-were cultured with extracts of salted Chinese cabbage, fresh kimchi and fermented kimchi(for 1 week) for 12 or 20 days. Control was supplemented with PBS(phosphate buffer saline) excluding kimchi extract. The results of spleen cell, bone marrow cell, and thymus cell cultures showed similar tendency: control medium accelerated death of cells, extracts of salted Chinese cabbage reduced the rate of cell death, and extracts of fresh kimchi and fermented kimchi promoted cell growth. From these results, it could be suggested that kimchi possibly has an effect on the hematopoietic ability and increases immune cell development and growth in vivo.

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